Instruction Manual for revvity models including: Celigo Image Cytometer, Image Cytometer, Cytometer
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DocumentDocumentEvery cell. Every well. Celigo® image cytometer · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Overview Celigo image cytometer overview The benchtop Celigo image cytometry system provides high-throughput, whole-well imaging and quantitative data through image analysis in brightfield and up to four fluorescent channels, for a wide variety of cell-based assays. It is routinely used to investigate adherent and suspension cells, 3D tumor spheroids and colonies of iPSC and cancer stem cells. It is compatible with microplates from 6 to 1536-well and T-flask formats. The workflow based intuitive software provides concurrent imaging and analysis; kinetic analysis such as time-lapse growth tracking, flow cytometry-like gating analysis and reporting of cell populations. Cell images of specific populations may be displayed with color overlays. The Celigo system allows users to perform high-speed, fully automated imaging and quantification of a wide range of cell types across complex sample types. It enables an extensive menu of applications including label-free cell counting, confluence-based cell growth tracking, killing assays, apoptosis, cell cycle analysis, migration and invasion assays, as well as cellular assays for receptor internalization, protein expression and detection, phosphorylation and phagocytosis. Core technology Proprietary optics Whole-well images are captured moving the plate once per well Illumination Flat illumination and excellent edge-to-edge contrast images every cell in every well. Image segmentation Brightfield and fluorescence algorithms for accurate segmentation of adherent and suspension cells Adherent cells Suspension cells For research use only. Not approved for diagnostic or therapeutic use. 2 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Whole-well, high resolution images acquired at high speed 1. Image entire plate 2. Analyze cell images Overview 3. Results data curves & cell images Image · Proprietary optical design enables uniform illumination and consistent edge contrast · Image and count every cell in each well: 0 - 100,000 cells/96-well · 5 imaging channels with brightfield and 4 fluorescent channels Analyze · Fast scanning for image acquisition and analysis with minimal plate movement ensuring minimal sample disruption · Accurately quantify cells and colonies with a non-invasive method · Measure adherent cells without trypsinization Results · Save experiment settings quickly run the same assay on many plates without additional set up · System automatically stitches multiple fields of view into a single full resolution image Imaging channels Imaging Channels BBrriigghhttfFieieldld 628/688 ex/em 531/629 ex/em 483/536 ex/em 337/470 ex/em Celigo easily integrates with robotic arms, plate stackers, automated incubators and liquid handlers. 3 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Applications Celigo applications Fluorescent assays Cell cycle, viability/cell health, internalization & phagocytosis, co-culture, surface proteins & antibodies, cell secretion, transfection/transduction, apoptosis, migration Cell counting Adherent cell counting, suspension cell counting, fluorescent cell counting, T-cells, splenocytes Fluorescent assays Brightfield Brightfield Adherent cell growth tracking, suspension cell counting, embryoid bodies, colonies, spheroids, wound healing, morphology Cell line development Single colony identification, single tumor sphere identification, single cell per well verification, transfection/transduction Cell Counting els reproIgPrSaCmming dCeveelllolipnme ent Migrati Virology Virology Viral titer, viral infection, antibody neutralization, transduction efficiency, cytopathic effect, CAR T cell-mediated cytotoxicity Migration | Invasion Chemotaxis, wound healing, transwell invasion, 3D migration, 3D invasion Image | Analyze | Results on | Invasion Immuno-oncology 3D mod Immuno-oncology Direct cell counting, visualization and documentation of all cells, using gating interface, ADCC, direct NK cell killing, CAR-T, CDC iPSC reprogramming Fibroblast doubling, iPSC colony counting, embryoid body formation, immunostaining for differentiation 3D models Growth inhibition, apoptosis, tumor spheroid viability, invasion into matrigel, migration onto ECM, tumorsphere formation & clonogenic survival, EBs & PDOs, 3D confrontation assay 4 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Integrated cell imaging and analysis in the same software Oncology · Apoptosis · Cell cycle · Cell viability · DNA synthesis · Cytotoxicity · Cell migration · Cell morphology · Immunophenotyping · Metabolism · Wound healing · Chemotaxis Immuno-oncology · Direct cell-mediated cytotoxicity · Antibody-dependent cell-mediated cytotoxicity (ADCC) · Complement-dependent cytotoxicity · Antibody-mediated cytotoxicity 3D tumor spheroid modeling · Growth tracking · Growth inhibition · Apoptosis | Viability · Invasion | Migration · Patient derived organoids (PDOs) Stem cell research · iPS cell line generation · Embryoid body morphology · Stem cell marker analysis Virology assays · Foci and plaque counting · Antibody neutralization · Hemagglutination assay Cell line development and biologics production · Single cell detection · Transfection | Transduction efficiency · CRISPR gene editing monitoring · Single cell to single colony monitoring · Growth tracking · Cell secretion · Hybridoma screening · Routine quality control Applications Cells counted to the well edge Propidium iodide and Hoechst stained 3D spheroid Surface marker stained iPS cells 5 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Application spotlights Immuno-oncology Accurate & consistent cell-mediated cytotoxicity assay for immuno-oncology The Celigo system images and counts cells directly within the experimental environment, while other methods often require the removal of cells from the experimental plate to perform end-point assays. Improve your data by performing consistent and accurate direct cell counts. Immuno-oncology assays additionally benefit from Celigo's increased sensitivity since cell death is measured at the individual cell level. Consequently, fewer target and effector cells are required, allowing for effective use of precious primary cell samples to generate the highest number of data points. Typically, only a few minutes are required for imaging and analyzing a 96- or 384-well plate.* Quantify killing & visualize formation of immune complexes: Direct cell counting assay in a 96-well plate A B C T = 0 hr 11,381 live tumor cells/well T = 6 hr: 1,635 live tumor cells/well Count every cell in every well Acquire whole-well, high resolution images (in brighfield and fluorescent) and count every cell in every well (A). Auto-save hi-res images Celigo images are used to verify results and are automatically saved for all wells (A). Kinetic measurement of tumor cell killing Images from the same 96-well plate (B, C) were acquired at 0 hours and 6 hours during a non-destructive, kinetic assay using calcein AM. Celigo images verify the decrease of calcein positive live tumor cells from 11,381 to 1,635 within the same well. *Somanchi SS, et al. PLoS ONE. 2015 Oct 10(10) Kummerow C, et al. Eur J Immunol. 2014 Jun 44(6):1870-2 6 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Antibody-dependent cell-mediated cytotoxicity (ADCC) assays NK cell mediated cytotoxicity effects on targeted cells Live calcein AM-positive cells were counted to determine the effect of a bimodal fusion protein (M7824) on NK cell-mediated cytotoxicity in co-cultures. Target cells (A549) were pretreated with TGF-1 and IFN- and stained with calcein AM (CAM). The NK cells were isolated from PBMCs and co-cultured with the A549 cells. · Cytotoxicity imaged in co-cultures · Percent specific lysis determined Application spotlight: immuno-oncology Untreated TGF-1 + IFN- % Specific lysis Fluorescent and bright field images of A549 treated with M7824 proteins. After 16 hours of co-culture with NK cells, propidium iodide (PI) was added, and live cells (CAM+/PI-) target cells were counted. M7824 (ng/mL) M7824 (ng/mL) Specific Lysis (%): Pretreatment with TGF-1 and IFN- sensitized A549 target cells to ADCC by M7824. Oncoimmunology 2017, 6(10):e1349589 7 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Tracking bispecific antibody induced cytotoxicity in complex co-culture Multi-valent antibody analysis (DuetMab Assay) The Celigo was used to directly count co-culture of prenatal cells and HER2 KO cells stained with CellTrackerTM Green and Violet, respectively. The assay investigated the selective cytotoxicity effect of the monovalent bispecific IgG (DuetMab) on a co-culture of prenatal cells and HER2 KO cells. Application spotlight: immuno-oncology 0 Hours Cell viabilities were calculated by direct cell counting, which showed differences in DuetMabs dose response between the prenatal and HER2 KO cell types (IC50: 1.08 nM and 10.71 nM, respectively). 72 Hours Fluorescent images of time 0 and 72 hours of prenatal and HER2 KO cells incubated with DuetMabs 8 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Application spotlight: drug screening Drug screening Discover combination therapies, investigate mechanisms of drug resistance and signaling pathways Fast and high-throughput cell counting assays are needed for drug discovery to satisfy the need to test an increasingly large number of cell models with large numbers of compounds and compound combinations. Gene editing technologies, such as CRISPR/Cas9, are rapidly expanding phenotypic in vitro cell models for oncology, and the success of immune and combination therapies has increased the number of compounds to be tested. · Count and analyze cells directly in experimental plates · Up to 5 imaging channels (e.g. BF, far red, red, green, blue) · Whole-well imaging for more accurate analysis · Direct high-throughput counting of suspension and adherent cells Label-free kinetic cytotoxicity assays Multi-drug growth inhibition · 96-well plate set-up with 4 compounds at 10 concentrations and a control · Scans the plates at multiple time points over hours or days · Automatic growth curve generation for each well Fu X, et al. Breast Cancer Res. 2014 Sep 11;16(5):430 Giuliano M, et al. Clin Cancer Res. 2015 Sep 1;21(17):3995-4003 9 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Automatically quantify 2D migration | invasion assays Image and quantify transwell-based chemotaxis, invasion and migration of suspension and adherent cells. Suspension cells located inside the insert (A) migrate through the porous membrane toward the chemoattractant in the bottom of the plate. Adherent cells located on the top of an ECM coated insert (C) invade and migrate to the bottom side of the membrane insert. Celigo automatically images and counts the migrated suspension and adherent cells (B, D). Image and quantify wound healing assays The entire well is imaged at multiple time points. Software automatically, and concurrently with imaging, calculates the cell confluence in each well. Application spotlight: drug screening Increase in Confluence Monitor growth inhibition using direct cell counting Dose-dependent growth inhibition Generated images are from two wells, B2 and B10, post drug treatment at doses of 33.3 m and 0.005 m respectively. · Visual determination of growth inhibition (greater in B2 than B10). · Identification and segmentation of individual cells (green) for each drug treatment. Well B2 Drug dose of 33.3 M Well B10 Drug dose of 0.005 M Well B2 Drug dose of 33.3 M Well B10 Drug dose of 0.005 M Bright field image Bright field image with single cell segmentation 10 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Application spotlight: 3D tumor models 3D tumor models Image cytometry enabling high-throughput 3D tumor spheroid screening 3D tumor spheroids are highly complex models increasingly used for cancer drug screening assays due to their better physiological relevance. Unlike 2D models, where cells can be enumerated and analyzed individually, the 3D model, which consists of clustered cells, requires a more advanced analysis method to characterize and quantify the effect of drugs on the entire spheroid. Therefore, to create a multi-parametric analysis platform, various orthogonal assays are required to screen the drug compounds. Celigo's high-throughput imaging and analysis is achieved by acquiring a single image from a single focal plane per well, which allows rapid screening of 96- and 384-well plates, typically in less than 2 minutes. Growth inhibition studies in microplates 1. 1. Form and treat 3D tumor spheroids Four days after seeding, spheroids were treated with compounds for 72 hours in u-bottom ultra low attachment 96-well plates. 2. Analyze 3D tumor spheroids Whole-well images are acquired and analyzed at multiple 2. days after adding drug compounds. Tumor sphere diameter under different drug treatments is exported into Excel for further data analysis. 11 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Application spotlight: 3D tumor models Real-time kinetic apoptosis and viability screening Measure the effects on apoptosis and viability with fluorescent image analysis. Tumor spheroids were treated with different drug compounds and screened to measure the effects on apoptosis and viability. The drug effects are characterized by measuring caspase 3/7 and propidium iodide (PI) fluorescent intensities over time. 12 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Tumor spheroid drug screening Visualize drug compound screening Spheroid growth, viability (calcein AM/ PI), apoptosis (Caspase 3-7/Hoechst) and invasion are analyzed. Brightfield and fluorescent images acquired by the Celigo visually confirm the state of the spheroid under different drug treatments. Drug screening data analysis Data generated from image analysis: Compound 3 and 4 were highly cytotoxic to the spheroids, while compound 5 and 12 showed cytostatic effects. Application spotlight: 3D tumor models 13 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Growth tracking and viability of organoids 3D organoid analysis Organoids are a 3D in vitro culture system derived from self-organizing stem cells. They can recapitulate the in vivo architecture, functionality, and genetic signature of original tissues and are thought to be a relevant model for evaluating drug effects. In the examples shown here, green segmentation is added to bright field images for quick and easy identification of Celigo counted organoids. Dome Method Viability determination Addition of non-perturbing dyes like propidium iodide (PI) generates valuable viability data, as seen in these whole-well images. Application spotlight: 3D tumor models Spin Method Perform organoid analysis in 24- to 384-well plates Whole-well imaging for organoid screening can be done in 24-well plates down to 384-well plates. Propidium Iodide Brightfield and PI Overlay 14 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Application spotlight: cell line development Cell line development through gene editing Quantify and monitor cell line development mediated by CRISPR/Cas9 system Chinese hamster ovary (CHO) cells are widely used as cell factories for the production of biologics. Gene editing technologies are used to alter function and to enhance production. During gene editing optimization and monitoring of stable clones, Celigo whole-well imaging allows for: · Direct cell counting and quantification of transfection/transduction efficiencies · Rapid high-throughput imaging · Direct documentation and verification of single cell to a single colony Monitor cell health and expansion during cell line development Quantify transfection efficiencies* Whole-well imaging combined with the 2D gating interface, allows high-throughput testing of transfection constructs and as a quality control measure during expansion to verify cell line stability. Monitor single cell to single colony formation Generated colonies are imaged and analyzed for colony number, size, shape and expression of a fluorescent marker indicative of a successful transfection. Monitor cell expansion during batch culturing During culture expansion, cell growth and viability were measured using a bright field and multi-channel fluorescent assay. (Images below BF = total cells, PI = dead cells, Hoechst = live cells) Brightfield Propidium Iodide Hoechst+ cells 47,380 PI+ cells 23,884 Viability 50.4% Hoechst *Grav LM, Lee JS, Gerling S, et al. (2015) One-step generation of triple knockout CHO cell lines using CRISPR/Cas9 and fluorescent enrichment. Biotechnol. J. 10, 1446-1456 Lee JS, Kallehauge TB, Pedersen LE, Kildegaard HF (2015) Site-specific integration in CHO cells mediated by CRISPR/Cas9 and homology-directed DNA repair pathway. Sci Rep. Feb 25;5 Hansen HG, Nilsson CN, Lund AM, et al. (2015) Versatile microscale screening platform for improving recombinant protein productivity in Chinese hamster ovary cells. Sci Rep. Dec, 11:5 15 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Monitor tumor sphere formation in microwell plates: single cell to a single sphere Single cell to single tumor sphere Use whole-well imaging to confirm a single cell per well (shown on Day 0) and verify its formation into a single tumor sphere (shown on Day 15). Day 0 Day 15 Identification of a single cell Formation of a single tumor sphere Quantification of tumor sphere size Each tumor sphere (shown at right) grew from a single cell clone seeded on day 1. · Images show the variation in formed tumor sphere size and morphology. · A size distribution graph was generated showing the variation of tumor sphere sizes. Application spotlight: cell line development Automated, high-throughput, characterization and differentiation of induced pluripotent stem cells Celigo image cytometer is utilized during the multi-stage process of iPSC generation. · Perform brightfield confluence and fluorescent viability assays. · Track fibroblast growth, monitor cell expansion during pre and post freeze/thaw cycles. · Image iPSC colonies using fluorescent fixed and live-cell markers for monitoring the re-programming process. · Examine differentiation by imaging and quantifying embryoid bodies and cell-line specific markers iPSC-derived cardiomyocytes stained with troponin-T and Hoechst. On the right, the same population of cells is analyzed using Celigo gating interface. Cells outlined in yellow represent troponin-T negative cells and those outlined in red are troponin-T positive differentiated cardiomyocytes. Paull D, et al. Nat Methods. 2015 Sep;12(9):885-92 Zhou H, et al. Stem Cell Rev. 2015 Aug;11(4):652-65 Proschel C, at al. EMBO Mol Med. 2014 Apr;6(4):504-18 16 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Application spotlight: virology Virology Analysis of viral-based assays for vaccine, therapy and disease research: automation, high-throughput and multi-parameters The Celigo imaging cytometer is an essential technology for the standardization and optimization of viral based assays. With rapid image acquisition and data analysis, viral plaque, antibody neutralization, and multiplexed high-throughput assays are achieved in minutes. Also, enhanced sensitivity, for measuring viral titers of low-level infections, is achieved through direct cell counts of whole-well images. The versatility of the Celigo instrument provides fast, efficient and reliable results for your virology research. Large dynamic range for viral titers via direct cell counting Count every infected cell to determine the viral titer HEK293T cell seeded at 80% confluence and infected with a 10-fold serial dilution of GFP-labeled lentivirus. Post infection, cells were imaged on the Celigo (outlined in red; Figure 1A), and the fluorescent cell count for each well was used to generate a viral titer curve (see Figure 1B) 1x Figure 1A 10x 100x Figure 1B Yang ML, et al. Sci Rep. 2017 Mar 6;7:43829 Randhawa P, et al. Antiviral Res. 2014 Dec 112:113-9 17 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Application spotlight: virology Increase throughput by automating plaque imaging and analysis Count brightfield, fluorescent and stained viral plaques Image and analyze plaques from cytolytic and non-cytopathic viruses: areas of infection in confluent cell culture indicated by horseradish peroxidase (HRP) for non-cytopathic viruses (Figure 2A) and zones of clearing for cytolytic viruses (Figure 2B). Early viral infections can be seen by detecting zones of viral infection (plaques) by detecting the GFP signal within the infected cells (Figure 2C). Figure 2A. Figure 2B. Figure 2c. Perform real-time and end point monitoring of viral infections Monitor the influenza virus infection Influenza infected cells were fixed, stained and imaged (Figure 3A and 3B). The software identifies and enumerates the number of infected cells (figure 3D), as well as total number of cells (Figure 3C). · Whole-well imaging allows for the identification of infected cells even at low viral titers. Figure 3A. Figure 3B. Figure 3C. Figure 3D. 18 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Celigo specifications Software Illumination/optics Fluorescent channels Proprietary image acquisition and processing software Powerful analysis software/computer workstation Microsoft Windows 1 LED-based enhanced brightfield imaging channel with uniform well illumination 4 LED-based fluorescent channels Proprietary F-theta lens with superior well edge-to-edge contrast Galvanometric mirrors for fast imaging of large areas Large chip CCD camera (2024x2024 pixels) 1, 2, 4 or 8 µm/pixel resolution Channel Blue Excitation 377/50 Dichroic 409 Green 483/32 509 Red 531/40 593 Plate compatibility High-speed imaging Weight and dimensions Power requirements Regulatory compliance Focusing modes Far-Red 628/40 660 6, 12, 24, 48, 96, 384, 1536 well plates (black, white and clear wall plates) T-25 and T-75 flasks Slides and cell array plate profiles available upon request Less than 2 minutes per 384-well plate Dimensions: 19.5 "W x 16 "H x 24 "D (49.5 cm x 40 cm x 61 cm) Weight: 117 lbs. (53 kg) 110-220 VAC 50-60 Hz CE marking Hardware-based auto focus, image-based auto focus, manual focus Emission 470/22 536/40 629/53 688/31 Specifications Typical dyes Hoechst, DAPI FITC, Calcein, GFP, AlexaFluor® 488 R-PE, PI, Texas Red, AlexaFluor® 568 DRAQ5®, AlexaFluor® 647 19 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Plate scan times Plate type 24-well plate 96-well plate 96-well plate 384-well plate 384-well plate Supported plate types Plate name 6-Well BD FalconTM 353046 plate 6-Well CorningTM 3516 plate 12-Well CorningTM 3513 plate 24-Well CorningTM 3524 plate 24-Well Revvity VisiplateTM 1450606 plate 96 Well-ULA Revvity ULA-96U-020 plate 96-Well CorningTM 3603 plate 96-Well GreinerTM 655090 plate 96-Well Greiner 675090 plate 384 Well-ULA Revvity ULA-384U-020 plate 384-Well CorningTM 3542 plate 384-Well CorningTM 3680 plate 384-Well CorningTM 3764 plate 1536-Well CorningTM 3838 plate Images / well 1 16 16 1 4 Resolution (m / pixels) 2 2 1 2 2 Manufacturer Corning Corning Corning Corning Revvity Revvity Corning Greiner Greiner Revvity Corning Corning Corning Corning Specifications Typical time < 7 minutes < 3.5 minutes < 5 minutes < 2 minutes < 7 minutes Well type Clear Clear Clear Clear Black Clear, round bottom Black, white Black, white White, half-area Clear, round bottom Low volume, black Clear Black, white Black, white 20 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Celigo platform The Celigo platform's modular design allows you to customize for your laboratory's needs. With automated microplate handling for either kinetic end-point analysis or time-point analysis, as well as complementary reagent kits, Celigo can improve your lab's work-flow and increase its capabilities. Hardware · Celigo BF Only · Celigo 5 Channel Software 5.0 with Project Mode. 1. Select project file 2. Load prepared plate & click run Acquisition, analysis, and data export steps are done automatically 3. Get Results · Report Templates: present data in customer defined layouts · Multiple FCS ExpressTM file export options · FCS and ICE file formats · Single file per plate or individual files per well · With images or without images in ICE file · Compression option · Auto open in templates · Security and Logging add-ons in FCS ExpressTM Stacker Automation · Up to 50 plate capacity · 15 sec transfer rate between plates · Set up experiment and run · Data exports automatically for each plate · Handles plates with or without lids · 6-well up to 1536-well plate formats · Fits on standard lab bench · Ideal for endpoint assays · Easy to add on to existing Celigo instruments Celigo Specific Reagents · Formulations designed for Celigo · Project files associated for each reagent kit Celigo platform 21 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Plate stacker Parameter Celigo only dimensions Celigo + stacker dimensions Electrical power Compressor (sold with stacker) Operating temperature Operating humidity Instrument shipping & storage temperature Instrument shipping & storage humidity Specification 19 inch wide x 29 inch long x 17 inch high (with plate holder extended) 25 inch wide x 44 inch long x 40 inch high (total) 5 Electrical outlets needed at 100-240 VAC 50/60 Hz (30 Amps Total) 1.2 CFM @ 90 PSI with 1 gallon tank 15 ºC to 25 ºC 10% to 90% RH, non-condensing -18 ºC to 65 ºC 10% to 90% RH, non-condensing Plate stacker 22 | · Overview · Applications · Application spotlights · Immuno-oncology · Drug screening · 3D tumor models · Cell line development · Virology · Specifications · Platform highlights · Plate stacker · Automation · Reagents · contact us Reagents ViaStainTM viability reagents · Perform in-plate staining and analysis of suspension and adherent cells · Measure percent viability and number live/dead cells Name AO / PI staining solution Calcein AM Calcein-AM / PI cell viability kit Calcein AM / PI / Hoechst viability kit Hoechst/PI viability kit Catalog # CS2-0106 CS1-0119 CSK-0118 CSK-V0006-1 CSK-V0005-1 ViaStainTM apoptosis reagents · Measure programmed cell death using various apoptosis detection reagents: Annexin V, Caspase 3/7-Live cell, Caspase 3 and Caspase 8 · Use Caspase 3/7 reagents for 2D and 3D cultures Name Live Caspase 3/7 Detection for 2D/3D Culture Live Caspase 3/7 Detection for 2D/3D Culture with Hoechst Catalog # CS1-V0002-1 CSK-V0003-1 Annexin V-FITC CS1-0114 ViaStainTM proliferation/tracer reagents · Fluorescent proliferation and cell-labeling dyes for cell identification, cell tracking and co-culture experiments Name CMFDA CFSE Tracer Blue Calcein AM Catalog # CS1-P0001-1 CS1-P0002-1 CS1-P0003-1 CS1-0119 Reagents ViaStainTM viability ViaStainTM apoptosis ViaStainTM proliferation/tracer 23 | www.revvity.com Revvity, Inc. 940 Winter Street Waltham, MA 02451 USA www.revvity.com For a complete listing of our global offices, visit www.revvity.com Copyright ©2023, Revvity, Inc. 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