Cellometer. Vision CBA. Image Cytometry System for 20µl Cell-Based Assays

Transcription

1 Cellometer Vision CBA Image Cytometry System for µl Cell-Based Apoptosis Cell Cycle and Others

2 Features of the Vision CBA Image Cytometry System All-in-One System Basic cell counting, primary cell viability, and cellbased assays. Dual-Fluorescence for Accurate Primary Cell No interference from red blood cells. Analyze bone marrow, peripheral blood, and cord blood without lysing. Unique Algorithms for Advanced Cell Analysis Determine concentration and viability of hepatocytes, adipocytes, and other sophisticated cell types. Fast Results Obtain cell images, counts, size measurements, viability calculations, and population data in < minutes. Bone Marrow Aspirate: bright field image and dual-fluorescence image showing live and dead nucleated cells present Simple Cell-Based Pre-qualified reagents Small µl sample size Simple, image-based analysis Pre-defined instrument settings -specific data templates Accurate, consistent results Advantages of Cellometer Image Cytometry Cell Imaging Visually check cell morphology Ensure only cells of interest are counted Archive and re-analyze cell images Export images for publication Non-Fluidic Platform Disposable counting chambers - no washing Compatible with fragile cells Maintenance-free Robust optics modules and LED light sources Proprietary Pattern-Recognition Software IQ/OQ Validation and GMP/GLP Accessories Count individual cells in clusters Installation Qualification reagents/protocol Count irregular-shaped cells Operational Qualification reagents/protocol Count cells based on size On-site IQ or OQ Performance Eliminate debris from cell counts GMP/GLP Software Module Primary Hepatocytes: bright field image Primary Adipocytes: bright field counted image

3 Cellometer Vision CBA Image Cytometry System for Cell-Based Vision CBA combines the simplicity of image cytometry with the power of flow analysis software to offer simple, accurate cell-based assays. Apoptosis Detect programmed cell death based on Annexin-V binding, Caspase activation, Chromatin condensation, or changes in mitochondrial membrane potential Aggresome Detection Detection Detect inclusion body formation in response to the accumulation of aggregating proteins Detect the breakdown of intra-cellular components by formation of autophagosomes and autolysosomes (special transport vesicles) Cell Cell Cycle Cycle Determine population distribution by cell cycle phase based on DNA content: resting/growth phase (G/G1), DNA replication phase (S), cell division phase (GM) Multidrug Multidrug Resistance Resistance (MDR) (MDR) Detect multidrug resistance based on activity of ABC transporter proteins and the removal of compounds from the cell Measure cell division based on reduction of original cytoplasmic protein content (and fluorescence intensity) in each generation Surface Surface Marker Marker Quantify specific cell populations based on surface marker expression (CD56+ NK cells, CD+ stem cells, etc.) Determine the efficiency of transfection based on CFP, GFP, mcherry, RFP, TdTomato, or YFP expression Measure the number, concentration, and percentage of live and dead cells based on membrane integrity and/or metabolic activity Validated Cell Types for Many Research Areas Adipocytes Monocytes Lymphocytes Clinical Immunology: PBMCs Diabetes / Obesity: Adipocytes Immunotherapy: Leukocytes Microbiology: Yeast (Vision x) Oncology: Cell Lines Regenerative Medicine: Stem Toxicology: Hepatocytes Transplantation: Nucleated Vaccine Development: Splenocytes Hepatocytes Optimized for Primary Cell Analysis PBMCs Epithelial Stem Dendritic Neural Keratinocytes Splenocytes Contact Nexcelom regarding your cell type

4 Simple, µl Cell-Based Growing Menu of Optimized Assasys Simple staining procedures User-friendly sample preparation Detect inclusion body formation in response to the accumulation of aggregating proteins Measure cell division based on reduction of original cytoplasmic protein content (and fluorescence intensity) in each generation Non-Fluidic System Determine population distribution by cell cycle phase based on DNA content: resting/growth phase (G/G1), DNA replication phase (S), cell division phase (GM) Determine the efficiency of transfection based on CFP, GFP, mcherry, RFP, TdTomato, or YFP expression Detect multidrug resistance based on activity of ABC transporter proteins and the removal of compounds from the cell Measure the number, concentration, and percentage of live and dead cells based on membrane integrity and/or metabolic activity Simple, maintenance-free operation Confirmation of counted cells Fluorescent cell counting Accurate results Regenerative Medicine: Stem Toxicology: Hepatocytes Transplantation: Nucleated Vaccine Development: Splenocytes L e ge nd 5 GFP 7.5% Count 11 PBMCs Splenocytes 1 1.9% 9 % % % Cellometer % 8 Flow Cytometer % [Nocodazole] (µg/ml) View Nexcelom Vision CBA Publications at: Cell Population % of Gated CV FL1 (intensity) 1.77% % Live 16.68% FL1 (intensity) Apoptotic Necrotic Debris Cell Poliferation: CFSE Day 1 Contact Nexcelom regarding your cell type Stem Keratinocytes 6 Dendritic GFP % 9 Neural Optimized for Primary Cell Analysis FL (intensity) 1 Epithelial Oncology: Cell Lines Concentration (^6 cells/ml) Total Sub G G/G S G/M *FCS Express Flow Cytometry software is a product of De Novo Software. Comprehensive data: images, graphs, tables Day Day 5 Day 6 Frequency Microbiology: Yeast (Vision x) Lymphocytes Hepatocytes 8 Individual Cellometer assays are designed to utilize specific optics modules for maximum performance and discrimination between fluorescence channels. Each Vision instrument accommodates two optics modules at one time. To change a module, users simply open the access panel at the rear of the instrument, depress the lever and remove the appropriate optics module, then insert the new one in its place. Standard modules are listed in the table below. Custom fluorescence optics modules are also available. Colored Subpopulation Plot Data Plot Gated for Fluorescent Protein Expression Cell Cycle PI Histogram of the Gated Population.NDat Immunotherapy: Leukocytes Cell Cycle: PI Diabetes / Obesity: Adipocytes 5% Apoptosis: Annexin V-FITC / PI GFP 6 Clinical Immunology: PBMCs User-Changeable Fluorescence Optics Modules* Multi-sample analysis options Adipocytes 656 6% Review data for cell cycle, proliferation, apoptosis and other cell-based assays; including correlation to traditional flow cytometry. Pre-set data layouts with user-adjustable gates Monocytes 59 Export to FCS Express* for Flow-Like Data Output Validated Cell Types for Many Research Areas 875 Figure 1. Cell cycle histogram following incubation with., and.1µg/ml Nocodazole Bright Field and Fluorescent Cell Images Multidrug Resistance (MDR) Multidrug Resistance (MDR) 57 Figure. Percent of cells arrested at G/M Phase Pre-defined instrument settings Quantify specific cell populations based on surface marker expression (CD56+ NK cells, CD+ stem cells, etc.) CellCell Cycle Cycle Consistent results No washing, clogging or daily calibration Surface Marker Surface Marker Detect the breakdown of intra-cellular components by formation of autophagosomes and autolysosomes (special transport vesicles) Correlation to Flow 9 Automatic counting and data calculation Aggresome Detection Aggresome Detection Nocodazole Dose Response Detect programmed cell death based on Annexin-V binding, Caspase activation, Chromatin condensation, or changes in mitochondrial membrane potential Pre-set analysis parameters 1 Apoptosis To demonstrate the Vision CBA Image Cytometry System for cell cycle analysis, Jurkat cells were incubated overnight with various concentrations of Nocodazole, a cell cycle-arresting drug. More than % of the cell population was arrested at the G/M phase following incubation with. µg/ml Nocodazole. Cellometer Vision CBA results showed excellent correlation to results obtained with the LSRII flow cytometer. Optimized reagent concentrations Vision CBA combines the simplicity of image cytometry with the power of flow analysis software to offer simple, accurate cell-based assays. Proven Results G/M Phase % Validated Cell-Based Kits Cellometer Vision CBA Image Cytometry System for Cell-Based Optics Module Fluorophores Nucleic Acid Stains Fluorescent Proteins VB-5- Ex: 75 nm Em: 5 nm AlexaFluor 5 DAPI Hoechst Hoechst 58 BFP CFP VB-55- Ex: 75 nm Em: 55 nm Calcein FITC AlexaFluor 88 AO (acridine orange, +DNA) SYTO 9, SYTO 1 GFP YFP VB Ex: 55 nm Em: 595 nm AlexaFluor 56 AlexaFluor 555, Cy PE (R-phycoerythrin) Rhodamine B AlexaFluor 67 7-AAD Nile Red AlexaFluor 67, Cy5 APC (allophycocyanin) SYTO Orange Ds Red RFP TdTomato AO (acridine orange, +RNA) SYTO Red Crimson VB-66-5 Ex: 5 nm Em: 66 nm CFSE Fluorescence Intensity (R.U.) time course involving B1 B cells. An increasing % of daughter cells exhibiting decreased fluorescence were observed on days, 5, and 6. 5 VB Ex: 6 nm Em: 695 nm *This table is a partial list of compatible fluorophores, nucleic acid stains, and fluorescent proteins. Please contact Nexcelom technical support regarding compatibility of other reagents. Sytox, AlexaFluor, and Cy are trademarks of Life Technologies.

5 Proven Results To demonstrate the Vision CBA Image Cytometry System for cell cycle analysis, Jurkat cells were incubated overnight with various concentrations of Nocodazole, a cell cycle-arresting drug. More than % of the cell population was arrested at the G/M phase following incubation with. µg/ml Nocodazole. Cellometer Vision CBA results showed excellent correlation to results obtained with the LSRII flow cytometer. Nocodazole Dose Response 9 57 Correlation to Flow 6% % G/M Phase % % % % Figure 1. Cell cycle histogram following incubation with., and.1µg/ml Nocodazole View Nexcelom Vision CBA Publications at: Review data for cell cycle, proliferation, apoptosis and other cell-based assays; including correlation to traditional flow cytometry. Cellometer % Flow Cytometer % [Nocodazole] (µg/ml) Figure. Percent of cells arrested at G/M Phase User-Changeable Fluorescence Optics Modules* Individual Cellometer assays are designed to utilize specific optics modules for maximum performance and discrimination between fluorescence channels. Each Vision instrument accommodates two optics modules at one time. To change a module, users simply open the access panel at the rear of the instrument, depress the lever and remove the appropriate optics module, then insert the new one in its place. Standard modules are listed in the table below. Custom fluorescence optics modules are also available. Optics Module Fluorophores Nucleic Acid Stains Fluorescent Proteins VB-5- Ex: 75 nm Em: 5 nm VB-55- Ex: 75 nm Em: 55 nm VB Ex: 55 nm Em: 595 nm VB-66-5 Ex: 5 nm Em: 66 nm VB Ex: 6 nm Em: 695 nm AlexaFluor 5 Calcein FITC AlexaFluor 88 AlexaFluor 56 AlexaFluor 555, Cy PE (R-phycoerythrin) Rhodamine B AlexaFluor 67 7-AAD Nile Red AlexaFluor 67, Cy5 APC (allophycocyanin) DAPI Hoechst Hoechst 58 AO (acridine orange, +DNA) SYTO 9, SYTO 1 SYTO Orange AO (acridine orange, +RNA) SYTO Red BFP CFP GFP YFP 57.5 Ds Red RFP TdTomato Crimson *This table is a partial list of compatible fluorophores, nucleic acid stains, and fluorescent proteins. Please contact Nexcelom technical support regarding compatibility of other reagents. Sytox, AlexaFluor, and Cy are trademarks of Life Technologies.

6 Features of the Vision CBA Image Cytometry System See for Yourself Why the Top Ten Pharmaceutical Companies Trust Cellometer All-in-One System Basic cell counting, primary cell viability, and cellbased assays. On-Site Demonstrations are a convenient way to evaluate the Vision CBA System. An experienced Applications Specialist will arrive at your lab for a hands-on session to test your cells and demonstrate the Vision CBA for your application. Dual-Fluorescence for Accurate Primary Cell No interference from red blood cells. Analyze bone marrow, peripheral blood, and cord blood without lysing. Technical Seminars are an excellent way to introduce Cellometer systems to a lab group or collaborators in different laboratories within an organization. A trained biologist will discuss and demonstrate the capabilities and advantages of Cellometer image cytometry for cell viability and cell-based assays. Unique Algorithms for Advanced Cell Analysis Determine concentration and viability of hepatocytes, adipocytes, and other sophisticated cell types. Cellometer Vision CBA Image Cytometry System for µl Cell-Based Schedule a FREE on-line demonstration, on-site demonstration or technical seminar with a Nexcelom Applications Specialist today. Call or info@nexcelom.com Fast Results Obtain cell images, counts, size measurements, viability calculations, and population data in < minutes. Cellometer Cell Counters, Cell Analysis Systems & Image Cytometry Nexcelom offers a wide range of Cellometer systems developed and optimized for specific applications and cell types. Simple Cell-Based Pre-qualified reagents Small µl sample size Apoptosis Simple, image-based analysis Pre-defined instrument settings -specific data templates Accurate, consistent results Bone Marrow Aspirate: bright field image and dual-fluorescence image showing live and dead nucleated cells present Simply Counted Cell Cycle Image Cytometer Which Cellometer is Right for Me? Features Automated Cell Counters Image Cytometers Mini Auto T Auto Auto 1 K Vision CBA Vision CBA (x) Objective Magnification 5 Cell Line Cultured Primary Advantages of Cellometer Image Cytometry Cell Imaging Non-Fluidic Platform Visually check cell morphology Disposable counting chambers - no washing Ensure only cells of interest are counted Compatible with fragile cells Archive and re-analyze cell images Maintenance-free Export images for publication Robust optics modules and LED light sources Proprietary Pattern-Recognition Software IQ/OQ Validation and GMP/GLP Accessories Count individual cells in clusters Installation Qualification reagents/protocol Count irregular-shaped cells Operational Qualification reagents/protocol Count cells based on size On-site IQ or OQ Performance Eliminate debris from cell counts GMP/GLP Software Module Primary Hepatocytes: bright field image Primary Adipocytes: bright field counted image Nexcelom products are for RESEARCH USE ONLY and are not approved for diagnostic or therapeutic use. Copyright 17 Nexcelom Bioscience LLC. All Rights Reserved Rev.E 1/17 Cell / Sample Type Algae Platelets Low Concentration Cell Lines Yeast (Clean Sample) Primary cells (Messy Sample*) PBMCs, Splenocytes, Stem Apoptosis (Annexin V-FITC/PI) Apoptosis (Caspase Activity) (CytoID-green) Cell (CFSE) Yeast (Messy Sample) Hepatocytes Adipocytes*** Cell-Based ** YFP RFP Mitochondrial Potential (JC-1) Multi-drug Resistance (ABC Transporter) Surface Marker Analysis Cell Cycle (PI) GFP Vitality (Calcein-AM/PI) Image Cytometry** * A messy sample is a heterogeneous sample containing unwanted cell types, such as red blood cells, in addition to the cells of interest. ** FCS Express license must be purchased in order to perform Cell Based or Image Cytometry analysis *** Cellometer CHT-PD slides are required for cells greater than 8µm in diameter Nous contacter Service Technique (ractifs et instrumentation) : tech@ozyme.fr Service Commande - Clients : commande@ozyme.fr and Others