Cellometer Vision CBA

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1 Features of the Vision CBA Image Cytometry System All-in-One System Basic cell counting, primary cell viability, and cellbased assays. See for Yourself Why the Top Ten Pharmaceutical Companies Trust Cellometer Dual-Fluorescence for Accurate Primary Cell No interference from red blood cells. Analyze bone marrow, peripheral blood, and cord blood without lysing. 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. 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. 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 cellbased assays. Simple Cell-Based Pre-qualified reagents Small µl sample size Simple, image-based analysis Pre-defined instrument settings -specific data templates Schedule a FREE on-line demonstration, on-site demonstration or technical seminar with a Nexcelom Applications Specialist today. Accurate, consistent results Bone Marrow Aspirate: bright field image and dual-fluorescence image showing live and dead nucleated cells present Call or info@nexcelom.com Advantages of Cellometer Image Cytometry Cell Imaging Non-Fluidic Platform 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 Visually check cell morphology 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 Rev.A 9/ Copyright Nexcelom Bioscience LLC. All Rights Reserved Cellometer Vision CBA For more information, visit Contact us at: Nexcelom Bioscience Merrimack Street, Building 9 Lawrence, MA 8, USA info@nexcelom.com Phone: Fax: Image Cytometry System for µl Cell-Based Apoptosis Cell and Others

2 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 Automatic counting and data calculation Non-Fluidic System Pre-defined instrument settings Quantify specific cell populations based on surface marker expression (CD+ NK cells, CD+ stem cells, etc.) CellCell Determine population distribution by cell cycle phase based on DNA content: resting/growth phase (G/G), 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 Fluorescent cell counting Accurate results Multi-sample analysis options Clinical Immunology: PBMCs Microbiology: Yeast (Vision x) Toxicology: Hepatocytes Transplantation: Nucleated Vaccine Development: Splenocytes Count 9 PBMCs Splenocytes Dendritic Contact Nexcelom regarding your cell type 8 Cell Population % of Gated CV Concentration (^ cells/ml) Total Sub G G/G...8 S.9.. G/M *FCS Express Flow Cytometry software is a product of De Novo Software. Comprehensive data: images, graphs, tables % 8 9 % % % Cellometer % 8 Flow Cytometer % [Nocodazole] (µg/ml) Figure. Percent of cells arrested at G/M Phase View Nexcelom Vision CBA Publications at:.77% GFP - VB-- 7.% Live.8% FL (intensity) Apoptotic Necrotic Debris Cell Poliferation: CFSE Day Keratinocytes.9% Stem Optimized for Primary Cell Analysis GFP +.% Oncology: Cell Lines Regenerative Medicine: Stem 8 Immunotherapy: Leukocytes L e ge nd Neural Hepatocytes Day Day Day Frequency Diabetes / Obesity: Adipocytes Control Epithelial 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 PI Histogram of the Gated Population.NXDat % Apoptosis: Annexin V-FITC / PI GFP Cell : PI Lymphocytes 9 User-Changeable Fluorescence Optics Modules* FL (intensity) Adipocytes 87 Review data for cell cycle, proliferation, apoptosis and other cell-based assays; including correlation to traditional flow cytometry. Confirmation of counted cells Pre-set data layouts with user-adjustable gates Monocytes 7 Export to FCS Express* for Flow-Like Data Output Validated Cell Types for Many Research Areas Correlation to Flow 9 Figure. Cell cycle histogram following incubation with.,., and.µg/ml Nocodazole Bright Field and Fluorescent Cell Images Consistent results No washing, clogging or daily calibration Detect the breakdown of intra-cellular components by formation of autophagosomes and autolysosomes (special transport vesicles) 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 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-- Ex: 7 nm Em: nm AlexaFluor DAPI Hoechst Hoechst 8 BFP CFP VB-- Ex: 7 nm Em: nm Calcein FITC AlexaFluor 88 AO (acridine orange, +DNA) SYTO 9, SYTO GFP YFP VB-9- Ex: nm Em: 9 nm AlexaFluor AlexaFluor, Cy PE (R-phycoerythrin) Rhodamine B AlexaFluor 7 7-AAD Nile Red AlexaFluor 7, Cy APC (allophycocyanin) SYTOX Orange 7. AO (acridine orange, +RNA) SYTOX Red Ds Red RFP TdTomato VB-- Ex: nm Em: nm CFSE Fluorescence Intensity (R.U.) time course involving B B cells. An increasing % of daughter cells exhibiting decreased fluorescence were observed on days,, and. VB-9- Ex: nm Em: 9 nm 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.

3 Features of the Vision CBA Image Cytometry System All-in-One System Basic cell counting, primary cell viability, and cellbased assays. See for Yourself Why the Top Ten Pharmaceutical Companies Trust Cellometer Dual-Fluorescence for Accurate Primary Cell No interference from red blood cells. Analyze bone marrow, peripheral blood, and cord blood without lysing. 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. 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. 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 cellbased assays. Simple Cell-Based Pre-qualified reagents Small µl sample size Simple, image-based analysis Pre-defined instrument settings -specific data templates Schedule a FREE on-line demonstration, on-site demonstration or technical seminar with a Nexcelom Applications Specialist today. Accurate, consistent results Bone Marrow Aspirate: bright field image and dual-fluorescence image showing live and dead nucleated cells present Call or info@nexcelom.com Advantages of Cellometer Image Cytometry Cell Imaging Non-Fluidic Platform 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 Visually check cell morphology 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 Rev.A 9/ Copyright Nexcelom Bioscience LLC. All Rights Reserved Cellometer Vision CBA For more information, visit Contact us at: Nexcelom Bioscience Merrimack Street, Building 9 Lawrence, MA 8, USA info@nexcelom.com Phone: Fax: Image Cytometry System for µl Cell-Based Apoptosis Cell and Others

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 Automatic counting and data calculation Non-Fluidic System Pre-defined instrument settings Quantify specific cell populations based on surface marker expression (CD+ NK cells, CD+ stem cells, etc.) CellCell Determine population distribution by cell cycle phase based on DNA content: resting/growth phase (G/G), 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 Fluorescent cell counting Accurate results Multi-sample analysis options Microbiology: Yeast (Vision x) PBMCs Toxicology: Hepatocytes Transplantation: Nucleated Vaccine Development: Splenocytes Splenocytes Dendritic Contact Nexcelom regarding your cell type FL (intensity).9% 8 Cell Population % of Gated CV FL (intensity).77% 7.% Live.8% FL (intensity) Apoptotic Necrotic Debris Cell Poliferation: CFSE Concentration (^ cells/ml) Total Sub G G/G...8 S.9.. G/M *FCS Express Flow Cytometry software is a product of De Novo Software. Comprehensive data: images, graphs, tables % 8 9 % % % Cellometer % 8 Flow Cytometer % [Nocodazole] (µg/ml) Figure. Percent of cells arrested at G/M Phase View Nexcelom Vision CBA Publications at: Day Keratinocytes Stem Optimized for Primary Cell Analysis GFP +.8% Oncology: Cell Lines Regenerative Medicine: Stem GFP 7.% Immunotherapy: Leukocytes 9 Neural Hepatocytes Day Day Day Frequency Diabetes / Obesity: Adipocytes Count Clinical Immunology: PBMCs L e ge nd Epithelial 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 PI Histogram of the Gated Population.NXDat % Apoptosis: Annexin V-FITC / PI GFP Cell : PI Lymphocytes 9 User-Changeable Fluorescence Optics Modules* Adipocytes 87 Review data for cell cycle, proliferation, apoptosis and other cell-based assays; including correlation to traditional flow cytometry. Confirmation of counted cells Pre-set data layouts with user-adjustable gates Monocytes 7 Export to FCS Express* for Flow-Like Data Output Validated Cell Types for Many Research Areas Correlation to Flow 9 Figure. Cell cycle histogram following incubation with., and.µg/ml Nocodazole Bright Field and Fluorescent Cell Images Consistent results No washing, clogging or daily calibration Detect the breakdown of intra-cellular components by formation of autophagosomes and autolysosomes (special transport vesicles) 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 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-- Ex: 7 nm Em: nm AlexaFluor DAPI Hoechst Hoechst 8 BFP CFP VB-- Ex: 7 nm Em: nm Calcein FITC AlexaFluor 88 AO (acridine orange, +DNA) SYTO 9, SYTO GFP YFP VB-9- Ex: nm Em: 9 nm AlexaFluor AlexaFluor, Cy PE (R-phycoerythrin) Rhodamine B AlexaFluor 7 7-AAD Nile Red AlexaFluor 7, Cy APC (allophycocyanin) SYTOX Orange 7. AO (acridine orange, +RNA) SYTOX Red Ds Red RFP TdTomato VB-- Ex: nm Em: nm CFSE Fluorescence Intensity (R.U.) time course involving B B cells. An increasing % of daughter cells exhibiting decreased fluorescence were observed on days,, and. VB-9- Ex: nm Em: 9 nm 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.

5 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 Automatic counting and data calculation Non-Fluidic System Pre-defined instrument settings Quantify specific cell populations based on surface marker expression (CD+ NK cells, CD+ stem cells, etc.) CellCell Determine population distribution by cell cycle phase based on DNA content: resting/growth phase (G/G), 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 Fluorescent cell counting Accurate results Multi-sample analysis options Microbiology: Yeast (Vision x) PBMCs Toxicology: Hepatocytes Transplantation: Nucleated Vaccine Development: Splenocytes Splenocytes Dendritic Contact Nexcelom regarding your cell type FL (intensity).9% 8 Cell Population % of Gated CV FL (intensity).77% 7.% Live.8% FL (intensity) Apoptotic Necrotic Debris Cell Poliferation: CFSE Concentration (^ cells/ml) Total Sub G G/G...8 S.9.. G/M *FCS Express Flow Cytometry software is a product of De Novo Software. Comprehensive data: images, graphs, tables % 8 9 % % % Cellometer % 8 Flow Cytometer % [Nocodazole] (µg/ml) Figure. Percent of cells arrested at G/M Phase View Nexcelom Vision CBA Publications at: Day Keratinocytes Stem Optimized for Primary Cell Analysis GFP +.8% Oncology: Cell Lines Regenerative Medicine: Stem GFP 7.% Immunotherapy: Leukocytes 9 Neural Hepatocytes Day Day Day Frequency Diabetes / Obesity: Adipocytes Count Clinical Immunology: PBMCs L e ge nd Epithelial 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 PI Histogram of the Gated Population.NXDat % Apoptosis: Annexin V-FITC / PI GFP Cell : PI Lymphocytes 9 User-Changeable Fluorescence Optics Modules* Adipocytes 87 Review data for cell cycle, proliferation, apoptosis and other cell-based assays; including correlation to traditional flow cytometry. Confirmation of counted cells Pre-set data layouts with user-adjustable gates Monocytes 7 Export to FCS Express* for Flow-Like Data Output Validated Cell Types for Many Research Areas Correlation to Flow 9 Figure. Cell cycle histogram following incubation with., and.µg/ml Nocodazole Bright Field and Fluorescent Cell Images Consistent results No washing, clogging or daily calibration Detect the breakdown of intra-cellular components by formation of autophagosomes and autolysosomes (special transport vesicles) 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 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-- Ex: 7 nm Em: nm AlexaFluor DAPI Hoechst Hoechst 8 BFP CFP VB-- Ex: 7 nm Em: nm Calcein FITC AlexaFluor 88 AO (acridine orange, +DNA) SYTO 9, SYTO GFP YFP VB-9- Ex: nm Em: 9 nm AlexaFluor AlexaFluor, Cy PE (R-phycoerythrin) Rhodamine B AlexaFluor 7 7-AAD Nile Red AlexaFluor 7, Cy APC (allophycocyanin) SYTOX Orange 7. AO (acridine orange, +RNA) SYTOX Red Ds Red RFP TdTomato VB-- Ex: nm Em: nm CFSE Fluorescence Intensity (R.U.) time course involving B B cells. An increasing % of daughter cells exhibiting decreased fluorescence were observed on days,, and. VB-9- Ex: nm Em: 9 nm 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 All-in-One System Basic cell counting, primary cell viability, and cellbased assays. See for Yourself Why the Top Ten Pharmaceutical Companies Trust Cellometer Dual-Fluorescence for Accurate Primary Cell No interference from red blood cells. Analyze bone marrow, peripheral blood, and cord blood without lysing. 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. 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. 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 cellbased assays. Simple Cell-Based Pre-qualified reagents Small µl sample size Simple, image-based analysis Pre-defined instrument settings -specific data templates Schedule a FREE on-line demonstration, on-site demonstration or technical seminar with a Nexcelom Applications Specialist today. Accurate, consistent results Bone Marrow Aspirate: bright field image and dual-fluorescence image showing live and dead nucleated cells present Call or info@nexcelom.com Advantages of Cellometer Image Cytometry Cell Imaging Non-Fluidic Platform 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 Visually check cell morphology 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 Rev.A 9/ Copyright Nexcelom Bioscience LLC. All Rights Reserved Cellometer Vision CBA For more information, visit Contact us at: Nexcelom Bioscience Merrimack Street, Building 9 Lawrence, MA 8, USA info@nexcelom.com Phone: Fax: Image Cytometry System for µl Cell-Based Apoptosis Cell and Others

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