{"id":38744,"date":"2023-09-19T14:00:51","date_gmt":"2023-09-19T12:00:51","guid":{"rendered":"https:\/\/phiab.com\/?p=38744"},"modified":"2023-11-01T11:04:38","modified_gmt":"2023-11-01T10:04:38","slug":"fluorescence-capture-protocol","status":"publish","type":"post","link":"https:\/\/phiab.com\/protocol\/fluorescence-capture\/","title":{"rendered":"Fluorescence Capture Protocol"},"content":{"rendered":"<div class=\"wp-block-group is-style-banner-image is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\"><figure class=\"wp-block-image size-large background-image is-landscape block-editor-block-list__block block-editor-block-list__first\" id=\"top\" style=\"--phi-media-width:1024px;--phi-media-height:341px;--phi-aspect-ratio:3.0029325513196;\">\n    <img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"Green fluorescent cells on a black background.\" class=\"wp-image-38672 lazyload figure__media\" width=\"1024\" height=\"341\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-banner-1024x341.jpg\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-banner-1024x341.jpg 1024w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-banner-300x100.jpg 300w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-banner-768x256.jpg 768w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-banner-150x50.jpg 150w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-banner-512x171.jpg 512w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-banner-1280x427.jpg 1280w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-banner.jpg 1536w\" data-sizes=\"auto\">\n  <\/figure>\n  <div class=\"wp-block-columns alignfull is-style-no-collapse is-layout-flex wp-container-core-columns-layout-1 wp-block-columns-is-layout-flex block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\"><div class=\"wp-block-column is-vertically-aligned-bottom is-style-default is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout is-empty block-editor-block-list__block block-editor-block-list__first\" style=\"flex-basis:15%;\"><\/div>\n    <div class=\"wp-block-column is-vertically-aligned-center is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second\" style=\"flex-basis:70%;\">\n      <h1 class=\"wp-block-heading alignfull has-text-align-center is-style-default has-white-color has-transparent-black-background-color has-text-color has-background has-huge-font-size block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" id=\"h-fluorescence-capture-protocol\" style=\"font-weight:600;\"><strong style=\"font-weight:bold;\">Fluorescence Capture<\/strong> <strong>Protocol<\/strong><\/h1>\n    <\/div>\n    <div class=\"wp-block-column is-vertically-aligned-center is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout is-empty block-editor-block-list__block block-editor-block-list__last\" style=\"flex-basis:15%;\"><\/div>\n  <\/div>\n<\/div>\n\n<div class=\"wp-block-media-text alignwide has-media-on-the-right is-stacked-on-mobile is-vertically-aligned-center floated-parent\" style=\"grid-template-columns:auto 20%;--phi-media-width:39px;--phi-media-height:39px;--phi-aspect-ratio:1;--phi-media-max-width:20%;\"><figure class=\"wp-block-media-text__media is-svg is-landscape floating-element\">\n    <img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"Three green cells and four blue cells in a circle.\" class=\"wp-image-38747 size-full lazyload figure__media\" data-media-id=\"38747\" width=\"39\" height=\"39\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/assay_fluorescence.svg\">\n  <\/figure><div class=\"wp-block-media-text__content block-editor-block-list__layout\"><p class=\"is-style-highlight wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">This protocol helps to set up a <a href=\"https:\/\/phiab.com\/holomonitor\/fluorescence\/\">Fluorescence Capture<\/a> using <a href=\"https:\/\/phiab.com\/holomonitor\/fluorescence\/\">HoloMonitor\u00ae M4FL<\/a> and the <a href=\"https:\/\/phiab.com\/holomonitor\/cell-imaging-software\/\">HoloMonitor\u00ae App Suite<\/a> software. The <a href=\"https:\/\/phiab.com\/holomonitor\/fluorescence\/\">HoloMonitor\u00ae Fluorescence Capture<\/a> provides holography time-lapse images for further analysis by any other <a href=\"https:\/\/phiab.com\/product\/holomonitor-app-suite-software\/\">HoloMonitor\u00ae App Suite<\/a> application and accompanying fluorescence images for <a href=\"https:\/\/phiab.com\/applications\/single-cell-tracking\/\">Single Cell Tracking<\/a> and <a href=\"https:\/\/phiab.com\/holomonitor\/cell-morphology-assay\/\">Cell Morphology<\/a> analysis.<\/p>\n    <div class=\"wp-block-buttons alignwide is-horizontal is-content-justification-center is-layout-flex wp-container-core-buttons-layout-1 wp-block-buttons-is-layout-flex block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\">\n      <div class=\"wp-block-button\">\n        <a class=\"wp-block-button__link wp-element-button\" href=\"https:\/\/phiab.com\/reports\/AppSuite-4.0\/AppSuite-protocol-fluorescence-capture.pdf\">Download this protocol<\/a>\n      <\/div>\n    <\/div>\n  <\/div>\n<\/div>\n\n<div class=\"wp-block-group is-style-curve is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <div class=\"wp-block-columns alignwide is-layout-flex wp-container-core-columns-layout-2 wp-block-columns-is-layout-flex block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\"><div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first\"><h2 class=\"wp-block-heading has-phi-blue-color has-text-color block-editor-block-list__block block-editor-block-list__first\" id=\"requirements\">Requirements:<\/h2>\n      <ul class=\"has-normal-font-size wp-block-list block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\"><li class=\"block-editor-block-list__block block-editor-block-list__first\"><a href=\"https:\/\/phiab.com\/holomonitor\/cell-imaging-system\/\">HoloMonitor\u00ae M4&nbsp;with M4FL<\/a>, placed in incubator<\/li>\n        <li class=\"block-editor-block-list__block block-editor-block-list__second\">\n          <a href=\"https:\/\/phiab.com\/holomonitor\/cell-imaging-software\/\">HoloMonitor\u00ae M4&nbsp;App Suite<\/a>\n        <\/li>\n        <li class=\"block-editor-block-list__block\">Culture vessel of choice with cells, including fluorescent cells for positive control<\/li>\n        <li class=\"block-editor-block-list__block\"><a href=\"https:\/\/phiab.com\/holomonitor\/hololids\/?swcfpc=1\">HoloLid\u2122 <\/a>for selected vessel<\/li>\n        <li class=\"block-editor-block-list__block block-editor-block-list__last\"><a href=\"https:\/\/phiab.com\/holomonitor\/accessories\/#vesselholders\">Vessel holder<\/a> for selected vessel<\/li>\n      <\/ul>\n    <\/div>\n    <div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second\"><h2 class=\"wp-block-heading has-phi-blue-color has-text-color block-editor-block-list__block block-editor-block-list__first\" id=\"output\">Output:<\/h2>\n      <ul class=\"has-normal-font-size wp-block-list block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\"><li class=\"block-editor-block-list__block block-editor-block-list__first\">Time-lapse images<\/li>\n        <li class=\"block-editor-block-list__block block-editor-block-list__second\">Time-lapse videos<\/li>\n        <li class=\"block-editor-block-list__block\">Fluorescence intensity values<\/li>\n        <li class=\"block-editor-block-list__block block-editor-block-list__last\">Selected application data analysis<\/li>\n      <\/ul>\n    <\/div>\n    <div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__last\"><h2 class=\"wp-block-heading has-phi-blue-color has-text-color block-editor-block-list__block block-editor-block-list__first\" id=\"reanalysis\">Reanalysis:<\/h2>\n      <ul class=\"is-style-default wp-block-list block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second\">\n        <li class=\"block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\">Guided assays<ul class=\"wp-block-list block-editor-block-list__layout\"><li class=\"block-editor-block-list__block block-editor-block-list__first\">\n              <a href=\"https:\/\/phiab.com\/holomonitor\/cell-qc-assay\/?swcfpc=1\">Cell Quality Control<\/a>\n            <\/li>\n            <li class=\"block-editor-block-list__block block-editor-block-list__second\">\n              <a href=\"https:\/\/phiab.com\/holomonitor\/cell-proliferation-assay\/\">Cell Proliferation<\/a>\n            <\/li>\n            <li class=\"block-editor-block-list__block\">\n              <a href=\"https:\/\/phiab.com\/applications\/kinetic-motility\/?swcfpc=1\">Kinetic Cell Motility<\/a>\n            <\/li>\n            <li class=\"block-editor-block-list__block block-editor-block-list__last\">\n              <a href=\"https:\/\/phiab.com\/applications\/dose-response\/?swcfpc=1\">Kinetic Dose Response<\/a>\n            <\/li>\n          <\/ul>\n        <\/li>\n      <\/ul>\n      <ul class=\"wp-block-list block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__last\">\n        <li class=\"block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\">In-depth analysis<ul class=\"wp-block-list block-editor-block-list__layout\"><li class=\"block-editor-block-list__block block-editor-block-list__first\">\n              <a href=\"https:\/\/phiab.com\/applications\/single-cell-tracking\/?swcfpc=1\">Single Cell Tracking<\/a>\n            <\/li>\n            <li class=\"block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\">\n              <a href=\"https:\/\/phiab.com\/holomonitor\/cell-morphology-assay\/?swcfpc=1\">Cell Morphology<\/a>\n            <\/li>\n          <\/ul>\n        <\/li>\n      <\/ul>\n    <\/div>\n  <\/div>\n<\/div>\n\n<div class=\"wp-block-group alignwide is-style-curve has-phi-blue-background-color has-background is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <h2 class=\"wp-block-heading alignwide is-style-page-break has-large-font-size block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" id=\"h-preparations\">\n    <strong>PREPARATIONS<\/strong>\n  <\/h2>\n<\/div>\n\n<h3 class=\"wp-block-heading alignwide has-small-large-font-size\" id=\"h-materials\">Materials<\/h3>\n\n<div class=\"wp-block-group alignwide is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <ul class=\"wp-block-list block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\"><li class=\"block-editor-block-list__block block-editor-block-list__first\"><strong><a href=\"https:\/\/phiab.com\/holomonitor\/?swcfpc=1\">HoloMonitor\u00ae M4FL<\/a><\/strong>, placed inside the incubator<\/li>\n    <li class=\"block-editor-block-list__block block-editor-block-list__second\"><a href=\"https:\/\/phiab.com\/holomonitor\/cell-imaging-software\/\">HoloMonitor\u00ae App Suite<\/a> software<\/li>\n    <li class=\"block-editor-block-list__block\"><strong>Cell culture vessel<\/strong>. Please check <a href=\"https:\/\/phiab.com\/protocol\/recommended-vessel-list\/\">our list<\/a> of recommended vessels.<\/li>\n    <li class=\"block-editor-block-list__block\"><strong>Cells<\/strong>, including a <strong>positive fluorescence control<\/strong><\/li>\n    <li class=\"block-editor-block-list__block\"><a href=\"https:\/\/phiab.com\/holomonitor\/hololids\/?swcfpc=1\">HoloLid\u2122<\/a> for the selected vessel<\/li>\n    <li class=\"block-editor-block-list__block\"><a href=\"http:\/\/phiab.com\/holomonitor\/accessories\/#vesselholders\">Vessel holder<\/a> for selected vessel<\/li>\n    <li class=\"block-editor-block-list__block block-editor-block-list__last\"><a href=\"https:\/\/phiab.com\/resources\/manuals-protocols\/\">Setup and Operation Manual<\/a> for HoloMonitor\u00ae<\/li>\n  <\/ul>\n<\/div>\n\n<h2 class=\"wp-block-heading alignwide has-small-large-font-size\" id=\"h-steps\">Steps<\/h2>\n\n<div class=\"wp-block-group alignwide is-style-restart-numbering is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\"><div class=\"wp-block-media-text alignwide has-media-on-the-right is-stacked-on-mobile is-vertically-aligned-center is-style-border is-style-lightbox floated-parent block-editor-block-list__block block-editor-block-list__first\" style=\"--phi-media-width:708px;--phi-media-height:474px;--phi-aspect-ratio:1.4936708860759;\"><figure class=\"wp-block-media-text__media is-landscape floating-element\">\n      <div class=\"lightbox-image-wrapper\">\n        <img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"calibration\" class=\"wp-image-31309 size-full lazyload figure__media\" data-media-id=\"31309\" width=\"708\" height=\"474\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2021\/10\/calibration.png\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2021\/10\/calibration.png 708w, https:\/\/phiab.com\/wp-content\/uploads\/2021\/10\/calibration-300x201.png 300w, https:\/\/phiab.com\/wp-content\/uploads\/2021\/10\/calibration-150x100.png 150w, https:\/\/phiab.com\/wp-content\/uploads\/2021\/10\/calibration-512x343.png 512w\" data-sizes=\"auto\" data-full-url=\"https:\/\/phiab.com\/wp-content\/uploads\/2021\/10\/calibration.png\">\n      <\/div>\n    <\/figure><div class=\"wp-block-media-text__content block-editor-block-list__layout\"><p class=\"is-style-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Seed the cells with about <strong>5&nbsp;% confluence<\/strong> (ca. 6000 \u2013 11000&nbsp;cells\/cm<sup>2<\/sup>).<br>\u25baPlease note that too few cells may lead to inadequate results due to auto-focus failure.<br>\u25baPlease note that a positive fluorescent control that<br>is representative of your exmeriment set up is essential.<\/p>\n      <p class=\"is-style-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\">Place the vessel in the incubator and let <strong>cells attach<\/strong> for 2-24&nbsp;hours.<\/p>\n      <p class=\"is-style-numbered wp-block-paragraph block-editor-block-list__block\"><strong>Start <\/strong>the <strong>software <\/strong>and wait for complete instrument <strong>initialization<\/strong>.<\/p>\n      <p class=\"is-style-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\">Run an<strong> auto-calibration<\/strong>. With successful calibration (as shown in the picture), the instrument is ready to use.<\/p>\n    <\/div>\n  <\/div>\n  <figure class=\"wp-block-table alignwide is-style-stripes block-editor-block-list__block block-editor-block-list__second\">\n    <table>\n      <tbody><tr><td>\n            <strong>Vessel<\/strong>\n          <\/td><td class=\"has-text-align-center\" data-align=\"center\">\n            <strong>Vendor cat.<br>number<\/strong>\n          <\/td><td class=\"has-text-align-center\" data-align=\"center\">\n            <strong>HoloLid\u2122<\/strong>\n          <\/td><td class=\"has-text-align-center\" data-align=\"center\">\n            <strong>Final working<br>volume<\/strong>\n          <\/td><td class=\"has-text-align-center\" data-align=\"center\">\n            <strong>Growth area,<br>cm2\/well<\/strong>\n          <\/td><td class=\"has-text-align-center\" data-align=\"center\">\n            <strong>Vessel cut in<br>a holder<\/strong>\n          <\/td>\n        <\/tr><tr><td>Sarstedt TC-dish 35<\/td><td class=\"has-text-align-center\" data-align=\"center\">83.3900<\/td><td class=\"has-text-align-center\" data-align=\"center\">71110<\/td><td class=\"has-text-align-center\" data-align=\"center\">3.0 mL\/well<\/td><td class=\"has-text-align-center\" data-align=\"center\">8.00<\/td><td class=\"has-text-align-center\" data-align=\"center\">NA<\/td>\n        <\/tr><tr><td>Sarstedt TC 6-well plate<\/td><td class=\"has-text-align-center\" data-align=\"center\">83.3920.005<\/td><td class=\"has-text-align-center\" data-align=\"center\">71120<\/td><td class=\"has-text-align-center\" data-align=\"center\">3.0 mL\/well<\/td><td class=\"has-text-align-center\" data-align=\"center\">8.80<\/td><td class=\"has-text-align-center\" data-align=\"center\">top left<\/td>\n        <\/tr><tr><td>Sarstedt lumox\u00ae 24-multiwell plate<\/td><td class=\"has-text-align-center\" data-align=\"center\">94.6000.014<\/td><td class=\"has-text-align-center\" data-align=\"center\">71130<\/td><td class=\"has-text-align-center\" data-align=\"center\">1.9 mL\/well<\/td><td class=\"has-text-align-center\" data-align=\"center\">1.90<\/td><td class=\"has-text-align-center\" data-align=\"center\">top left<\/td>\n        <\/tr><tr><td>Sarstedt lumox\u00ae 96-multiwell plate<\/td><td class=\"has-text-align-center\" data-align=\"center\">94.6000.024<\/td><td class=\"has-text-align-center\" data-align=\"center\">71140<\/td><td class=\"has-text-align-center\" data-align=\"center\">170 \u00b5L\/well<\/td><td class=\"has-text-align-center\" data-align=\"center\">0.34<\/td><td class=\"has-text-align-center\" data-align=\"center\">top left<\/td>\n        <\/tr><tr><td>ibidi\u00ae \u03bc-dish 35 mm, high<\/td><td class=\"has-text-align-center\" data-align=\"center\">81156<\/td><td class=\"has-text-align-center\" data-align=\"center\">71111<\/td><td class=\"has-text-align-center\" data-align=\"center\">2.5 mL\/well<\/td><td class=\"has-text-align-center\" data-align=\"center\">3.50<\/td><td class=\"has-text-align-center\" data-align=\"center\">NA<\/td>\n        <\/tr><tr><td>ibidi\u00ae \u03bc-plate 24 Well Black<\/td><td class=\"has-text-align-center\" data-align=\"center\">80241<\/td><td class=\"has-text-align-center\" data-align=\"center\">71131<\/td><td class=\"has-text-align-center\" data-align=\"center\">2.5 mL\/well<\/td><td class=\"has-text-align-center\" data-align=\"center\">1.90<\/td><td class=\"has-text-align-center\" data-align=\"center\">NA<\/td>\n        <\/tr><tr><td>Eppendorf CCCadvanced\u00ae FN1 &#8211; 6 well<\/td><td class=\"has-text-align-center\" data-align=\"center\">0038110010<\/td><td class=\"has-text-align-center\" data-align=\"center\">71150<\/td><td class=\"has-text-align-center\" data-align=\"center\">3.0 mL\/well<\/td><td class=\"has-text-align-center\" data-align=\"center\">9.40<\/td><td class=\"has-text-align-center\" data-align=\"center\">bottom right<\/td>\n        <\/tr>\n      <\/tbody>\n    <\/table>\n  <\/figure>\n  <p class=\"is-style-numbered wp-block-paragraph block-editor-block-list__block\">Sterilize the <strong>HoloLids\u2122<\/strong> according to the specific <a href=\"https:\/\/phiab.com\/resources\/manuals-protocols\/#accessories\">HoloLid\u2122 protocol<\/a>.<\/p>\n  <p class=\"is-style-numbered wp-block-paragraph block-editor-block-list__block\">Add the <strong>treatment <\/strong>to your cells. The final working volumes per well, essential for using <strong>HoloLids\u2122<\/strong>, are shown in the table above.<\/p>\n  <p class=\"is-style-numbered wp-block-paragraph block-editor-block-list__block\"><strong>Slide <\/strong>the cell culture <strong>vessel <\/strong>onto the Vessel <strong>holder<\/strong>, its grips facing towards you. Ensure that the vessel is parallel to the holder. There is a spring that holds the vessel in place.<br>\u25baWhen using multi-well plates, place them with the cut-off corner to the left.<\/p>\n  <p class=\"is-style-numbered wp-block-paragraph block-editor-block-list__block\"><strong>Replace <\/strong>the standard lids with the <strong>HoloLid\u2122<\/strong>.<\/p>\n  <p class=\"is-style-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\"><strong>Put <\/strong>the vessel <strong>holder <\/strong>with the sample on the <strong>HoloMonitor\u00ae M4&nbsp;stage<\/strong>.<\/p>\n<\/div>\n\n<div class=\"wp-block-group alignwide is-style-curve has-phi-blue-background-color has-background is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <h2 class=\"wp-block-heading alignwide is-style-page-break has-large-font-size block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" id=\"h-calibrating-fluorescence-signal-in-live-view\">\n    <strong>CALIBRATING FLUORESCENCE SIGNAL IN LIVE VIEW<\/strong>\n  <\/h2>\n<\/div>\n\n<h3 class=\"wp-block-heading alignwide is-style-numbered has-small-large-font-size\" id=\"h-focus-holography-image\">Focus Holography image<\/h3>\n\n<div class=\"wp-block-group alignwide is-style-restart-numbering is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <div class=\"wp-block-columns alignwide are-vertically-aligned-top is-style-border is-style-lightbox is-layout-flex wp-container-core-columns-layout-3 wp-block-columns-is-layout-flex block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\"><div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first\" style=\"flex-basis:50%;\"><p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Go to <strong>Live View Tab<\/strong> on the left side panel.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\">Select the correct<strong> vessel map<\/strong> from the drop-down list.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Find a position with cells. Preferably, the position should be as centered in the vessel as possible.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\">In <strong>Controls<\/strong>, make sure that the holography image of the cells is <strong>in focus<\/strong> by adjusting the <strong>Z-position<\/strong> of the stage using the up\/down arrows. Ensure that <strong>Autofocus<\/strong> is ticked and then adjust the <strong>Z-position<\/strong> until the Software focus bar shows <strong>1300\u00b15<\/strong>.<br>\u25ba The <strong>Z-position<\/strong> will be saved with the calibration and used as the default <strong>Z-position<\/strong> during <strong>Capture Setup<\/strong>.<\/p>\n    <\/div>\n    <div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\" style=\"flex-basis:50%;\">\n      <figure class=\"wp-block-image is-style-lightbox has-caption is-landscape block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" style=\"--phi-media-width:1024px;--phi-media-height:546px;--phi-aspect-ratio:1.8754578754579;\">\n        <div class=\"lightbox-image-wrapper\">\n          <img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"Screenshot of HoloMonitor App Suite showing the fluorescence calibration screen.\" class=\"wp-image-38814 lazyload figure__media\" width=\"1024\" height=\"546\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_hol-1024x546.jpg\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_hol-1024x546.jpg 1024w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_hol-300x160.jpg 300w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_hol-768x409.jpg 768w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_hol-150x80.jpg 150w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_hol-512x273.jpg 512w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_hol-1536x818.jpg 1536w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_hol-1280x682.jpg 1280w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_hol.jpg 1554w\" data-sizes=\"auto\" data-full-url=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_hol.jpg\">\n        <\/div><figcaption class=\"wp-element-caption\">Live View window, Controls tab<\/figcaption>\n      <\/figure>\n    <\/div>\n  <\/div>\n<\/div>\n\n<h3 class=\"wp-block-heading alignwide is-style-numbered has-small-large-font-size\" id=\"h-calibrate-fluorescence-focus-plane\">Calibrate Fluorescence Focus Plane<\/h3>\n\n<div class=\"wp-block-group alignwide is-style-restart-numbering is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <div class=\"wp-block-columns alignwide are-vertically-aligned-top is-style-border is-style-lightbox is-layout-flex wp-container-core-columns-layout-4 wp-block-columns-is-layout-flex block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\"><div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first\" style=\"flex-basis:50%;\"><p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Go to the <strong>Fluorescence <\/strong>tab.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\">Turn on <strong>Fluorescence live capture<\/strong> by pressing the <strong>Play<\/strong> button (or take a <strong>Single<\/strong> image repeatedly) while manually adjusting the <strong>Fluorescence focus<\/strong> until you get a fluorescence signal.<br>\u25ba You can use the arrow keys on the keyboard.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Check that the view is set to mixed channel.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Set <strong>Exposure time<\/strong> and <strong>Gain<\/strong>. For more information see <a href=\"https:\/\/phiab.com\/resources\/manuals-protocols\/#manuals\">HoloMonitor\u00ae M4FL Setup and Operation Manual<\/a>.<br>\u25ba Increasing <strong>gain <\/strong>value might be beneficial when the fluorescence signal is weak, and App Suite struggles to find focus automatically.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Press <strong>Autofocus <\/strong>to automatically set best focus offset.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">If the software fails to find optimal focus, adjust it manually by adjusting the <strong>fluorescence focus<\/strong> slider.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Press <strong>Calibrate<\/strong>.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Use the <strong>Position<\/strong>, <strong>Scale <\/strong>and <strong>Rotation <\/strong>controls to adjust and align the fluorescence overlay to fit the holographic image.<br>\u25ba This is an iterative step. Thus, position, scale, and rotation adjustments do not need to be performed sequentially.<br>\u25ba If needed, you can decrease the Step size to make finer adjustments. You can also change the opacity of the fluorescence overlay to compare it with the holographic image better.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Turn Off the <strong>Fluorescence Live Capture<\/strong>.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Press <strong>Save calibration<\/strong> when the fluorescence focus and alignment are as good as possible.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\">Go to the <strong>Apps <\/strong>tab and select the <strong>Fluorescence Capture Application <\/strong>and proceed by clicking the <strong>Setup <\/strong>button.<\/p>\n    <\/div>\n    <div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\" style=\"flex-basis:50%;\"><figure class=\"wp-block-image size-large is-style-lightbox has-caption is-landscape block-editor-block-list__block block-editor-block-list__first\" style=\"--phi-media-width:1024px;--phi-media-height:584px;--phi-aspect-ratio:1.7534246575342;\">\n        <div class=\"lightbox-image-wrapper\">\n          <img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"Screenshot from App Suite showing the fluorescence calibration.\" class=\"wp-image-38828 lazyload figure__media\" width=\"1024\" height=\"584\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_FL-1024x584.jpg\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_FL-1024x584.jpg 1024w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_FL-300x171.jpg 300w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_FL-768x438.jpg 768w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_FL-150x86.jpg 150w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_FL-512x292.jpg 512w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_FL-1280x730.jpg 1280w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_FL.jpg 1355w\" data-sizes=\"auto\" data-full-url=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-calibration_FL.jpg\">\n        <\/div><figcaption class=\"wp-element-caption\">Live View window, Fluorescence tab<\/figcaption>\n      <\/figure>\n      <p class=\"is-style-default is-style-round-corners is-style-lighter is-style-highlight has-phi-light-grey-background-color has-background wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\">Fluorescence signal intensity can be evaluated using the measure tool (or histogram), which are found to the right of the Live View image.<\/p>\n      <p class=\"is-style-default is-style-round-corners is-style-lighter is-style-highlight has-phi-light-grey-background-color has-background wp-block-paragraph block-editor-block-list__block\">To reduce the fluorescent exposure during calibration, turn off fluorescence live capture. Instead, take a single fluorescence image every time you need to confirm a change during the calibration.<\/p>\n      <div class=\"wp-block-group is-style-round-corners is-style-highlight is-style-set-numbering has-phi-light-grey-background-color has-background is-layout-constrained wp-block-group-is-layout-constrained block-editor-block-list__layout block-editor-block-list__block\"><p class=\"is-style-lighter is-style-default wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">If the arrows in Controls for Microscope Stage were used in<br>step 1.3&nbsp;to navigate to the location used for calibration, the<br>holographic position might need to be corrected:<\/p>\n        <p class=\"is-style-lighter is-style-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\">Go to Controls.<\/p>\n        <p class=\"is-style-numbered is-style-lighter wp-block-paragraph block-editor-block-list__block\">Turn OFF the Holographic Phase channel by p ressing the Stop button .<\/p>\n        <p class=\"is-style-numbered is-style-lighter wp-block-paragraph block-editor-block-list__block\">Press Correct Pos.<\/p>\n        <p class=\"is-style-numbered is-style-lighter wp-block-paragraph block-editor-block-list__block\">Turn ON the Holographic Phase channel by pressing the Play button.<\/p>\n        <p class=\"is-style-numbered is-style-lighter wp-block-paragraph block-editor-block-list__block\">Turn OFF the Holographic Phase channel by pressing the Stop button.<\/p>\n        <p class=\"is-style-numbered is-style-lighter wp-block-paragraph block-editor-block-list__block\">Take Single Fluorescence Capture.<\/p>\n        <p class=\"is-style-numbered is-style-lighter wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\">Check if Holographic Channel and Fluorescence Channel images are aligned. Repeat steps if needed.<\/p>\n      <\/div>\n      <figure class=\"wp-block-image size-large is-style-lightbox has-caption is-landscape block-editor-block-list__block block-editor-block-list__last\" style=\"--phi-media-width:1024px;--phi-media-height:611px;--phi-aspect-ratio:1.6759410801964;\">\n        <div class=\"lightbox-image-wrapper\">\n          <img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"Screenshot from App Suite showing buttons representing all available assays.\" class=\"wp-image-38829 lazyload figure__media\" width=\"1024\" height=\"611\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Front-page_FC-1024x611.jpg\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Front-page_FC-1024x611.jpg 1024w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Front-page_FC-300x179.jpg 300w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Front-page_FC-768x458.jpg 768w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Front-page_FC-150x90.jpg 150w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Front-page_FC-512x306.jpg 512w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Front-page_FC-1280x764.jpg 1280w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Front-page_FC.jpg 1397w\" data-sizes=\"auto\" data-full-url=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Front-page_FC.jpg\">\n        <\/div><figcaption class=\"wp-element-caption\">App Suite main window with selected Fluorescence Capture<\/figcaption>\n      <\/figure>\n    <\/div>\n  <\/div>\n<\/div>\n\n<div class=\"wp-block-group alignwide is-style-curve has-phi-blue-background-color has-background is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <h2 class=\"wp-block-heading alignwide is-style-page-break has-large-font-size block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" id=\"h-experiment-setup\">\n    <strong>EXPERIMENT SETUP<\/strong>\n  <\/h2>\n<\/div>\n\n<h3 class=\"wp-block-heading alignwide is-style-numbered has-small-large-font-size\" id=\"h-basic-setup-describe-the-experiment-and-assign-treatments-to-the-wells\">Basic setup: Describe the experiment and assign treatments to the wells<\/h3>\n\n<div class=\"wp-block-group alignwide is-style-restart-numbering is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <div class=\"wp-block-columns alignwide are-vertically-aligned-top is-style-border is-style-lightbox is-layout-flex wp-container-core-columns-layout-5 wp-block-columns-is-layout-flex block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\"><div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first\" style=\"flex-basis:50%;\"><p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Enter the experiment <strong>name<\/strong>, optional experiment <strong>description, <\/strong>and cell types.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\">Select the correct <strong>vessel map <\/strong>from the drop-down list.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Map <strong>treatments <\/strong>on the vessel map. <strong>Select wells<\/strong> by<br>marking them with the left mouse button while moving<br>the cursor over the relevant well(s).<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Add the<strong> treatment name(s)<\/strong> in the text box below the vessel map and click <strong>Add<\/strong> \/ press Enter. It is possible to<br>add wells as <strong>individual treatments<\/strong>. Marked well(s) are light blue, selected wells will appear dark blue.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\">Proceed to<strong> Capture setup<\/strong>.<\/p>\n    <\/div>\n    <div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\" style=\"flex-basis:50%;\">\n      <figure class=\"wp-block-image is-style-lightbox has-caption is-landscape block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" style=\"--phi-media-width:1024px;--phi-media-height:546px;--phi-aspect-ratio:1.8754578754579;\">\n        <div class=\"lightbox-image-wrapper\">\n          <img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"Screenshot from App Suite showing the basic Fluorescence Capture setup.\" class=\"wp-image-38866 lazyload figure__media\" width=\"1024\" height=\"546\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Basic-Setup-1024x546.jpg\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Basic-Setup-1024x546.jpg 1024w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Basic-Setup-300x160.jpg 300w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Basic-Setup-768x409.jpg 768w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Basic-Setup-150x80.jpg 150w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Basic-Setup-512x273.jpg 512w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Basic-Setup-1280x682.jpg 1280w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Basic-Setup.jpg 1360w\" data-sizes=\"auto\" data-full-url=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Basic-Setup.jpg\">\n        <\/div><figcaption class=\"wp-element-caption\">Basic Setup window<\/figcaption>\n      <\/figure>\n    <\/div>\n  <\/div>\n<\/div>\n\n<h3 class=\"wp-block-heading alignwide is-style-numbered has-small-large-font-size\" id=\"h-capture-setup-set-experiment-time-settings-choose-and-validate-capture-positions\">Capture setup: Set experiment time settings, choose and validate capture positions<\/h3>\n\n<div class=\"wp-block-group alignwide is-style-set-numbering is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <div class=\"wp-block-columns alignwide are-vertically-aligned-top is-style-border is-style-lightbox is-layout-flex wp-container-core-columns-layout-6 wp-block-columns-is-layout-flex block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\"><div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first\" style=\"flex-basis:50%;\"><p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Adjust the default settings for <strong>duration <\/strong>and <strong>interval<\/strong>.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\"><strong>Set <\/strong>Fluorescence capture <strong>interval<\/strong>.<br>\u25ba Captures set too often will induce phototoxicity.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Setup capture positions.<\/p>\n      <div class=\"wp-block-group is-style-restart-numbering is-layout-constrained wp-block-group-is-layout-constrained block-editor-block-list__layout block-editor-block-list__block\"><p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Setting capture locations:<\/p>\n        <div class=\"wp-block-group is-style-restart-numbering is-layout-constrained wp-block-group-is-layout-constrained block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second\"><p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Use the software <strong>automatically <\/strong>pre-set positions, change the number of positions if needed by pressing the <strong>Regenerate pattern<\/strong> button.<\/p>\n          <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\">If automatically generated positions are not suitable, find and set the positions <strong>manually<\/strong> (see <a href=\"#manual-setup\">Capture Setup: Manually choose and validate capture positions<\/a> section).<\/p>\n        <\/div>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block\">Setting Holographic focus:<br>\u25ba This step can be skipped if the Holographic <strong>Software focus<\/strong> was set manually to <strong>1300\u00b15<\/strong>.<\/p>\n        <div class=\"wp-block-group is-style-restart-numbering is-layout-constrained wp-block-group-is-layout-constrained block-editor-block-list__layout block-editor-block-list__block\"><p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Go to the<strong> Holography<\/strong> tab.<\/p>\n          <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\"><strong>Automatically<\/strong> optimize the Z-position by pressing the <strong>Start Optimization button<\/strong>. The software automatically tries to set a Z-value, where the <strong>Software focus<\/strong> would be as close as possible to <strong>1300\u00b15<\/strong>. Values for the inspected positions are displayed in the <strong>Position list<\/strong>.<br>\u25ba This function also validates the image quality. For more information about image quality, consult <a href=\"https:\/\/phiab.com\/resources\/manuals-protocols\/#manuals\">App Suite Setup and Operation Manual<\/a>.<\/p>\n          <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\">If the Automatically set <strong>Z-value<\/strong> does not lead to a <strong>Software focus<\/strong> of <strong>1300\u00b15<\/strong>, adjust the <strong>Z-value manually<\/strong> (see <a href=\"#manual-setup\">Capture Setup: Manually choose and validate capture positions<\/a> section).<\/p>\n        <\/div>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block\">Setting Fluorescence focus:<\/p>\n        <div class=\"wp-block-group is-style-restart-numbering is-layout-constrained wp-block-group-is-layout-constrained block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__last\"><p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Go to the Fluorescence tab.<\/p>\n          <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\">To automatically adjust the Fluorescence focus value for selected positions that have fluorescence signal at the beginning of the capture or all positions, press <strong>Run on Selected<\/strong> or <strong>Run on All<\/strong> under <strong>Autofocus Pre-experiment<\/strong>.<br>\u25ba This step can be skipped for positions\/wells where the fluorescence signal appears later during the capture.<\/p>\n          <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block\">To <strong>manually <\/strong>adjust the Fluorescence focus value for the selected position or all positions, see <em>Capture Setup: Manually choose and validate capture positions<\/em> section.<\/p>\n          <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\">Enable <strong>Continuous Refocus during experiment<\/strong> for the software to adjust Fluorescence focus during the capture.<\/p>\n        <\/div>\n      <\/div>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Adjust the fluorescence <strong>exposure time<\/strong> and <strong>gain <\/strong>if needed in the <strong>Fluorescence tab<\/strong>.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Ensure that the <strong>storage requirement<\/strong> for the experiment does not exceed the computer capacity.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\">When satisfied with the experiment setup, click <strong>Proceed<br>to Capture<\/strong>.<\/p>\n    <\/div>\n    <div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\" style=\"flex-basis:50%;\"><figure class=\"wp-block-image size-large is-style-lightbox has-caption is-portrait block-editor-block-list__block block-editor-block-list__first\" style=\"--phi-media-width:918px;--phi-media-height:1024px;--phi-aspect-ratio:0.896484375;\">\n        <div class=\"lightbox-image-wrapper\">\n          <img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"Screenshot from App Suite showing the Fluorecence Capture setup.\" class=\"wp-image-38868 lazyload figure__media\" width=\"918\" height=\"1024\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capture-Setup-918x1024.jpg\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capture-Setup-918x1024.jpg 918w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capture-Setup-269x300.jpg 269w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capture-Setup-768x857.jpg 768w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capture-Setup-150x167.jpg 150w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capture-Setup-512x571.jpg 512w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capture-Setup-1280x1428.jpg 1280w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capture-Setup.jpg 1357w\" data-sizes=\"auto\" data-full-url=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capture-Setup.jpg\">\n        <\/div><figcaption class=\"wp-element-caption\">Capture Setup window with opened steup tabs<\/figcaption>\n      <\/figure>\n      <p class=\"is-style-default is-style-round-corners is-style-lighter is-style-highlight has-phi-light-grey-background-color has-background wp-block-paragraph block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\">For the best Fluorescence imaging result, the Holographic Image Software focus value should be as close as possible to 1300\u00b15. Adjusting the Actual Z-value for each position might be necessary.<\/p>\n    <\/div>\n  <\/div>\n<\/div>\n\n<h3 class=\"wp-block-heading alignwide is-style-indented has-small-large-font-size\" id=\"manual-setup\">Capture setup: Manually choose and validate capture position<\/h3>\n\n<div class=\"wp-block-group alignwide is-style-set-numbering is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <div class=\"wp-block-columns alignwide are-vertically-aligned-top is-style-border is-style-lightbox is-layout-flex wp-container-core-columns-layout-7 wp-block-columns-is-layout-flex block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\"><div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first\" style=\"flex-basis:50%;\"><p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Setting capture locations:<\/p>\n      <div class=\"wp-block-group is-style-restart-numbering is-layout-constrained wp-block-group-is-layout-constrained block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second\"><p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Click the <strong>positions list<\/strong> button to view the list.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\">Press the <strong>Select all<\/strong> button and <strong>Delete <\/strong>to remove automatically generated positions.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block\">Press <strong>Add Current Location<\/strong> to add Fluorescence calibration position if wanted.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block\">Find a position with cells using arrows in <strong>Stage Controls<\/strong> or by clicking on the vessel map.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block\">Adjust the <strong>Z-value<\/strong> to the position where the <strong>Software focus<\/strong> is <strong>1300\u00b15<\/strong>.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block\">If satisfied, press <strong>Add current location<\/strong>.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\"><strong>Repeat<\/strong> until satisfied.<\/p>\n      <\/div>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Setting Holographic focus:<\/p>\n      <div class=\"wp-block-group is-style-restart-numbering is-layout-constrained wp-block-group-is-layout-constrained block-editor-block-list__layout block-editor-block-list__block\"><p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\"><strong>Visit <\/strong>the position.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\">Change position <strong>Z-value<\/strong> using Focus arrows in <strong>Stage Controls<\/strong>. Once the <strong>Software focus<\/strong> is <strong>1300\u00b15<\/strong>, press <strong>Apply Current Focus<\/strong>.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\">If satisfied, proceed to the next position. Otherwise, find a new location.<\/p>\n      <\/div>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Setting Fluorescence focus:<\/p>\n      <div class=\"wp-block-group is-style-restart-numbering is-layout-constrained wp-block-group-is-layout-constrained block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__last\"><p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Go to the <strong>Fluorescence<\/strong> tab.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\">Start <strong>Fluorescence live<\/strong> feed.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block\">Use <strong>Fluorescence focus<\/strong> slider to achieve the sharpest fluorescence image.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block\">Once satisfied, press <strong>Apply to Selected<\/strong> or <strong>Apply to All<\/strong>.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block\">Repeat for other positions if needed until satisfied.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block\">Enable <strong>Continuous Refocus during experiment<\/strong> for the software to adjust Fluorescence focus during the capture.<\/p>\n        <p class=\"is-style-box-numbered is-style-legal-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\">Click Proceed to capture.<\/p>\n      <\/div>\n    <\/div>\n    <div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\" style=\"flex-basis:50%;\">\n      <figure class=\"wp-block-image size-large is-style-lightbox has-caption is-portrait block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" style=\"--phi-media-width:918px;--phi-media-height:1024px;--phi-aspect-ratio:0.896484375;\">\n        <div class=\"lightbox-image-wrapper\">\n          <img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"Screenshot from App Suite showing the manual Fluorescence Capture setup procedure.\" class=\"wp-image-38882 lazyload figure__media\" width=\"918\" height=\"1024\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_manual_Capture-Setup-918x1024.jpg\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_manual_Capture-Setup-918x1024.jpg 918w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_manual_Capture-Setup-269x300.jpg 269w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_manual_Capture-Setup-768x857.jpg 768w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_manual_Capture-Setup-150x167.jpg 150w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_manual_Capture-Setup-512x571.jpg 512w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_manual_Capture-Setup-1280x1428.jpg 1280w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_manual_Capture-Setup.jpg 1357w\" data-sizes=\"auto\" data-full-url=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_manual_Capture-Setup.jpg\">\n        <\/div><figcaption class=\"wp-element-caption\">Capture Setup window with opened steup tabs<\/figcaption>\n      <\/figure>\n    <\/div>\n  <\/div>\n<\/div>\n\n<h3 class=\"wp-block-heading alignwide is-style-numbered has-small-large-font-size\" id=\"h-capture-review-the-experiment-in-real-time-during-the-time-lapse\">Capture: Review the experiment in real-time during the time-lapse<\/h3>\n\n<div class=\"wp-block-group alignwide is-style-restart-numbering is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <div class=\"wp-block-columns alignwide are-vertically-aligned-top is-style-border is-style-lightbox is-layout-flex wp-container-core-columns-layout-8 wp-block-columns-is-layout-flex block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\"><div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first\" style=\"flex-basis:50%;\"><p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Click <strong>Start Capture<\/strong>.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\">To stop the experiment ahead of time, click the <strong>Stop Capture<\/strong> button.<br>\u25ba It is not possible to restart the experiment once it has been stopped.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">To pause the experiment, click the <strong>Pause <\/strong>button. Press <strong>Resume <\/strong>button to continue the experiment.<br>\u25ba Missed frames can be appended to the end of the experiment or skipped.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">Click on <strong>Preview Results<\/strong> to preview the captured images during the run.<br>\u25baWait for the experiment to fully finish before starting data analysis.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\">When the Experiment capture finishes, click the <strong>Show Result<\/strong> button to go directly to the <strong>Experiment Overview<\/strong> tab.<\/p>\n    <\/div>\n    <div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\" style=\"flex-basis:50%;\">\n      <figure class=\"wp-block-image size-large is-style-lightbox has-caption is-landscape block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" style=\"--phi-media-width:1024px;--phi-media-height:536px;--phi-aspect-ratio:1.910447761194;\">\n        <div class=\"lightbox-image-wrapper\">\n          <img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"Screenshot from App Suite showing the Fluorescence Capture screen.\" class=\"wp-image-38880 lazyload figure__media\" width=\"1024\" height=\"536\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capturing-1-1024x536.jpg\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capturing-1-1024x536.jpg 1024w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capturing-1-300x157.jpg 300w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capturing-1-768x402.jpg 768w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capturing-1-150x79.jpg 150w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capturing-1-512x268.jpg 512w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capturing-1-1280x670.jpg 1280w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capturing-1.jpg 1412w\" data-sizes=\"auto\" data-full-url=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FC_Capturing-1.jpg\">\n        <\/div><figcaption class=\"wp-element-caption\">Capture window<\/figcaption>\n      <\/figure>\n    <\/div>\n  <\/div>\n<\/div>\n\n<div class=\"wp-block-group alignwide is-style-curve has-phi-blue-background-color has-background is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <h2 class=\"wp-block-heading alignwide is-style-page-break has-large-font-size block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" id=\"h-results-analysis\">\n    <strong>RESULTS &amp; ANALYSIS<\/strong>\n  <\/h2>\n<\/div>\n\n<h3 class=\"wp-block-heading alignwide is-style-indented has-small-large-font-size\" id=\"h-experiments-tab\">Experiments tab<\/h3>\n\n<div class=\"wp-block-group alignwide is-style-restart-numbering is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <div class=\"wp-block-columns alignwide are-vertically-aligned-top is-style-border is-style-lightbox is-layout-flex wp-container-core-columns-layout-9 wp-block-columns-is-layout-flex block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\"><div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first\" style=\"flex-basis:50%;\"><p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">Click Experiments to see a <strong>list of the experiments<\/strong>.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\">Click on the experiment title to open an <strong>experiment summary<\/strong>.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\">Click <strong>Open <\/strong>or double-click left mouse button to open the results page.<\/p>\n    <\/div>\n    <div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\" style=\"flex-basis:50%;\">\n      <figure class=\"wp-block-image size-large is-style-lightbox has-caption is-landscape block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" style=\"--phi-media-width:1024px;--phi-media-height:546px;--phi-aspect-ratio:1.8754578754579;\">\n        <div class=\"lightbox-image-wrapper\">\n          <img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"A screenshot from App Suite showing the Experiments overview.\" class=\"wp-image-38885 lazyload figure__media\" width=\"1024\" height=\"546\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Experiment-overview_FC-1024x546.jpg\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Experiment-overview_FC-1024x546.jpg 1024w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Experiment-overview_FC-300x160.jpg 300w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Experiment-overview_FC-768x409.jpg 768w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Experiment-overview_FC-150x80.jpg 150w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Experiment-overview_FC-512x273.jpg 512w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Experiment-overview_FC-1280x682.jpg 1280w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Experiment-overview_FC.jpg 1360w\" data-sizes=\"auto\" data-full-url=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/Experiment-overview_FC.jpg\">\n        <\/div><figcaption class=\"wp-element-caption\">Experiments tab<\/figcaption>\n      <\/figure>\n    <\/div>\n  <\/div>\n<\/div>\n\n<h3 class=\"wp-block-heading alignwide is-style-indented has-small-large-font-size\" id=\"h-experiment-overview-tab\">Experiment overview tab<\/h3>\n\n<div class=\"wp-block-group alignwide is-style-restart-numbering is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <div class=\"wp-block-columns alignwide are-vertically-aligned-center is-style-border is-style-lightbox is-layout-flex wp-container-core-columns-layout-10 wp-block-columns-is-layout-flex block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\"><div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first\" style=\"flex-basis:50%;\"><p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">See the experiment <strong>summary<\/strong>, view all <strong>images <\/strong>and go to the experiment <strong>setup<\/strong>.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\">Generate <strong>In-depth analysis<\/strong> data (fluorescence green area (\u03bcm2), fluorescence green average, fluorescence green max, fluorescence green min, fluorescence green std, and fluorescence green sum) from the captured images by clicking on the <strong>Cell Morphology<\/strong> or <strong>Single Cell Tracking<\/strong> icon. A new window for the In-depth analysis will open.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\"><strong>Create New Guided Assay Results<\/strong> from this experiment by clicking the button.<br>\u25ba Note that only the DHM images will be analysed using the guided assay analysis.<\/p>\n    <\/div>\n    <div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\" style=\"flex-basis:50%;\">\n      <figure class=\"wp-block-image size-large has-caption is-landscape block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" style=\"--phi-media-width:1024px;--phi-media-height:545px;--phi-aspect-ratio:1.8788990825688;\"><img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"A screenshot from App Suite showing the Experiment overview of a fluorescent experiment.\" class=\"wp-image-38886 lazyload figure__media\" width=\"1024\" height=\"545\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_FC-1024x545.jpg\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_FC-1024x545.jpg 1024w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_FC-300x160.jpg 300w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_FC-768x409.jpg 768w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_FC-150x80.jpg 150w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_FC-512x273.jpg 512w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_FC-1280x681.jpg 1280w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_FC.jpg 1360w\" data-sizes=\"auto\"><figcaption class=\"wp-element-caption\">Experiment overview tab<\/figcaption>\n      <\/figure>\n    <\/div>\n  <\/div>\n<\/div>\n\n<h3 class=\"wp-block-heading alignwide is-style-indented has-small-large-font-size\" id=\"h-in-depth-analysis\">In-depth analysis<\/h3>\n\n<div class=\"wp-block-group alignwide is-style-float-right is-style-restart-numbering is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout floated-parent\"><div class=\"wp-block-group is-style-default is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first floating-element\">\n    <figure class=\"wp-block-image alignleft size-large is-style-lightbox has-caption is-portrait block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" style=\"--phi-media-width:854px;--phi-media-height:1024px;--phi-aspect-ratio:0.833984375;\">\n      <div class=\"lightbox-image-wrapper\">\n        <img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"Shreenshot from App Suite showing the Identify cells windows.\" class=\"wp-image-38887 lazyload figure__media\" width=\"854\" height=\"1024\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL_Manual-changes-854x1024.jpg\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL_Manual-changes-854x1024.jpg 854w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL_Manual-changes-250x300.jpg 250w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL_Manual-changes-768x921.jpg 768w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL_Manual-changes-150x180.jpg 150w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL_Manual-changes-512x614.jpg 512w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL_Manual-changes-1280x1534.jpg 1280w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL_Manual-changes.jpg 1359w\" data-sizes=\"auto\" data-full-url=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL_Manual-changes.jpg\">\n      <\/div><figcaption class=\"wp-element-caption\">Identify cells window<\/figcaption>\n    <\/figure>\n  <\/div>\n  <div class=\"wp-block-columns is-layout-flex wp-container-core-columns-layout-11 wp-block-columns-is-layout-flex block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\">\n    <div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" style=\"flex-basis:100%;\">\n      <ul class=\"is-style-checkmark wp-block-list block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\"><li class=\"block-editor-block-list__block block-editor-block-list__first\">The first step for <strong>Cell Morphology<\/strong> analysis and <strong>Single Cell Tracking<\/strong> is to identify the cells.<\/li>\n        <li class=\"block-editor-block-list__block block-editor-block-list__second\">The software automatically calculates cell count and confluence for each image.<\/li>\n        <li class=\"block-editor-block-list__block\">Review the image quality and include\/ exclude images from results analysis. See the <strong>Image guide<\/strong> for more information.<\/li>\n        <li class=\"block-editor-block-list__block\">In the<strong> Identify cells<\/strong> under <strong>Fluorescence tab<\/strong>, adjust <strong>Holographic<\/strong> and <strong>Fluorescence Image overlay<\/strong> automatically or manually for individual frames, selected frames, or all frames.<\/li>\n        <li class=\"block-editor-block-list__block block-editor-block-list__last\">Follow corresponding <strong>Cell Morphology<\/strong> or <strong>Single Cell Tracking<\/strong> application protocols for data analysis.<\/li>\n      <\/ul>\n    <\/div>\n  <\/div>\n<\/div>\n\n<div class=\"wp-block-group alignwide is-style-curve has-phi-blue-background-color has-background is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <h2 class=\"wp-block-heading alignwide is-style-page-break has-large-font-size block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" id=\"h-one-experiment-multiple-results\" style=\"text-transform:uppercase;\">\n    <strong>One experiment \u2014 multiple results<\/strong>\n  <\/h2>\n<\/div>\n\n<ul class=\"is-style-checkmark wp-block-list block-editor-block-list__layout\">\n  <li class=\"block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\">This section helps to reanalyze data between different assays using HoloMonitor\u00ae App Suite software.<\/li>\n<\/ul>\n\n<h3 class=\"wp-block-heading alignwide has-small-large-font-size\" id=\"h-generating-guided-assay-results\">Generating Guided Assay results<\/h3>\n\n<div class=\"wp-block-group alignwide is-style-restart-numbering is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <div class=\"wp-block-columns alignwide are-vertically-aligned-top is-style-lightbox is-layout-flex wp-container-core-columns-layout-12 wp-block-columns-is-layout-flex block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\"><div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first\" style=\"flex-basis:50%;\"><p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">In the <strong>Experiment overview page<\/strong> under Guided Assay Results select <strong>Create New Result<\/strong>.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second\"><strong>Choose the type <\/strong>of analysis in the pop-up window and <strong>name <\/strong>the new result.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block\">If there are previous analyses it is possible to tick <strong>copy image analysis from<\/strong> and <strong>select <\/strong>the experiment to copy from. This will copy the image analysis settings from the selected result including all changes.<br>\u25ba For further data analysis steps, please see the respective <a href=\"https:\/\/phiab.com\/resources\/manuals-protocols\/#protocols\">assay protocol<\/a>.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__last\"><strong>Press create<\/strong>.<\/p>\n    <\/div>\n    <div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\" style=\"flex-basis:50%;\">\n      <figure class=\"wp-block-image has-caption is-landscape block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" style=\"--phi-media-width:1024px;--phi-media-height:545px;--phi-aspect-ratio:1.8788990825688;\"><img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"Screenshot from App Suite showing the re-analysis promt.\" class=\"wp-image-38889 lazyload figure__media\" width=\"1024\" height=\"545\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_reanalysis_-1024x545.jpg\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_reanalysis_-1024x545.jpg 1024w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_reanalysis_-300x160.jpg 300w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_reanalysis_-768x409.jpg 768w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_reanalysis_-150x80.jpg 150w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_reanalysis_-512x273.jpg 512w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_reanalysis_-1280x681.jpg 1280w, https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/overview_reanalysis_.jpg 1360w\" data-sizes=\"auto\"><figcaption class=\"wp-element-caption\">Experiment overview tab<\/figcaption>\n      <\/figure>\n    <\/div>\n  <\/div>\n<\/div>\n\n<h3 class=\"wp-block-heading alignwide has-small-large-font-size\" id=\"generating-in-depth-assay-results\">Generating In-depth Assay results<\/h3>\n\n<div class=\"wp-block-group alignwide is-style-restart-numbering is-layout-flow wp-block-group-is-layout-flow block-editor-block-list__layout\">\n  <div class=\"wp-block-columns alignwide are-vertically-aligned-top is-style-border is-style-lightbox is-layout-flex wp-container-core-columns-layout-13 wp-block-columns-is-layout-flex block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\"><div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__first\" style=\"flex-basis:46%;\"><p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__first\">In the <strong>Experiment overview page select the In-depth assay icon for the <\/strong>wanted result.<\/p>\n      <p class=\"is-style-box-numbered wp-block-paragraph block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\"><strong>Follow <\/strong>the respective assay protocol.<\/p>\n    <\/div>\n    <div class=\"wp-block-column is-layout-flow wp-block-column-is-layout-flow block-editor-block-list__layout block-editor-block-list__block block-editor-block-list__second block-editor-block-list__last\" style=\"flex-basis:54%;\">\n      <figure class=\"wp-block-image size-medium_large is-landscape block-editor-block-list__block block-editor-block-list__first block-editor-block-list__last\" style=\"--phi-media-width:959px;--phi-media-height:720px;--phi-aspect-ratio:1.3319444444444;\">\n        <a href=\"https:\/\/phiab.com\/holomonitor\/live-cell-assays\/?swcfpc=1\" class=\"figure__media figure__media-link\"><img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/phiab.com\/wp-content\/themes\/phi-xxvi\/images\/placeholder.svg\" alt=\"Cell proliferation assay protocol reanalysis\" class=\"wp-image-31545 lazyload\" data-media-id=\"31545\" width=\"959\" height=\"720\" data-src=\"https:\/\/phiab.com\/wp-content\/uploads\/2021\/10\/reanalysis-5.jpg\" data-srcset=\"https:\/\/phiab.com\/wp-content\/uploads\/2021\/10\/reanalysis-5.jpg 959w, https:\/\/phiab.com\/wp-content\/uploads\/2021\/10\/reanalysis-5-300x225.jpg 300w, https:\/\/phiab.com\/wp-content\/uploads\/2021\/10\/reanalysis-5-768x577.jpg 768w, https:\/\/phiab.com\/wp-content\/uploads\/2021\/10\/reanalysis-5-150x113.jpg 150w, https:\/\/phiab.com\/wp-content\/uploads\/2021\/10\/reanalysis-5-512x384.jpg 512w\" data-sizes=\"auto\"><\/a>\n      <\/figure>\n    <\/div>\n  <\/div>\n<\/div>\n\n<div class=\"wp-block-buttons is-horizontal is-content-justification-center is-layout-flex wp-container-core-buttons-layout-2 wp-block-buttons-is-layout-flex\">\n  <div class=\"wp-block-button\">\n    <a class=\"wp-block-button__link wp-element-button\" href=\"https:\/\/phiab.com\/reports\/AppSuite-4.0\/AppSuite-protocol-fluorescence-capture.pdf\">Download this protocol<\/a>\n  <\/div>\n<\/div>","protected":false},"excerpt":{"rendered":"The Fluorescence Capture allows HoloMonitor M4FL to capture fluorescent images for further analysis in other HoloMonitor App Suite assays. Requires App Suite 4.0 or higher.","protected":false},"author":8,"featured_media":38824,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"ub_ctt_via":"","footnotes":""},"categories":[6157],"tags":[],"featured_image_src":"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-swoosh.jpg","author_info":{"display_name":"Andreas Svensson","author_link":"https:\/\/phiab.com\/author\/ans\/"},"acf":[],"yoast_head":"<!-- This site is optimized with the Yoast SEO Premium plugin v21.6 (Yoast SEO v21.6) - https:\/\/yoast.com\/wordpress\/plugins\/seo\/ -->\n<title>HoloMonitor\u00ae Fluorescence Capture Protocol - Phase Holographic Imaging<\/title>\n<meta name=\"description\" content=\"This Fluorescence Capture protocol helps to set up fluorescence experiments using HoloMonitor\u00ae M4FL and the HoloMonitor\u00ae App Suite software.\" \/>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/phiab.com\/protocol\/fluorescence-capture\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Fluorescence Capture Protocol\" \/>\n<meta property=\"og:description\" content=\"This Fluorescence Capture protocol helps to set up fluorescence experiments using HoloMonitor\u00ae M4FL and the HoloMonitor\u00ae App Suite software.\" \/>\n<meta property=\"og:url\" content=\"https:\/\/phiab.com\/protocol\/fluorescence-capture\/\" \/>\n<meta property=\"og:site_name\" content=\"PHI\" \/>\n<meta property=\"article:published_time\" content=\"2023-09-19T12:00:51+00:00\" \/>\n<meta property=\"article:modified_time\" content=\"2023-11-01T10:04:38+00:00\" \/>\n<meta property=\"og:image\" content=\"https:\/\/phiab.com\/wp-content\/uploads\/2023\/09\/FL-swoosh.jpg\" \/>\n\t<meta property=\"og:image:width\" content=\"1536\" \/>\n\t<meta property=\"og:image:height\" content=\"673\" \/>\n\t<meta property=\"og:image:type\" content=\"image\/jpeg\" \/>\n<meta name=\"author\" content=\"Andreas Svensson\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:label1\" content=\"Written by\" \/>\n\t<meta name=\"twitter:data1\" content=\"Andreas Svensson\" \/>\n\t<meta name=\"twitter:label2\" content=\"Est. reading time\" \/>\n\t<meta name=\"twitter:data2\" content=\"12 minutes\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\/\/schema.org\",\"@graph\":[{\"@type\":\"WebPage\",\"@id\":\"https:\/\/phiab.com\/protocol\/fluorescence-capture\/\",\"url\":\"https:\/\/phiab.com\/protocol\/fluorescence-capture\/\",\"name\":\"HoloMonitor\u00ae Fluorescence Capture Protocol - 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