产品介绍
The Vision CBA System combines the simplicity of image cytometry with the power of flow analysis software to offer user-friendly cell-based assays, featuring:
Fluorescent Cell-Based Assays |
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Vision CBA
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Apoptosis (Annexin V-FITC / PI) | ||
Apoptosis (Caspase Activity) |
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Autophagy (CytoID-green) |
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Cell Proliferation (CFSE) |
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Cell Cycle (PI) | ||
GFP, RFP, YFP Transfection |
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Mitochondrial Potential / Early Cell Death (JC-1) | ||
Multi-Drug Resistance (ABC Transporter) |
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Surface Marker Analysis (FITC) |
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Viability (AO/PI, Trypan blue) |
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Vitality (Calcein-AM / PI) |
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Vision CBA and FCS Express Software enable automated data output to pre-set data layouts for individual cell-based assays. Users have the option to modify the automatic gating with instant visual display and data update.
Data layouts can be modified to accommodate multi-sample analysis and create customized analysis and / or quality control reports.
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Fluorescent dyes that stain DNA can be used to identify nucleated cells in a mixed cell population. Acridine orange (AO) is a nuclear staining (nucleic acid binding) dye permeable to both live and dead cells. It stains all nucleated cells to generate green fluorescence.
Fluorescent membrane-exclusion dyes that stain DNA can be used to identify dead nucleated cells in a mixed cell population. Propidiom iodide (PI) is a nuclear staining dye that can only enter cells with compromised membranes. Healthy cells exclude the dye. Dead nucleated cells stained with both AO and PI fluoresce red due to fluorescence resonance energy transfer.
Red blood cells seen in the bright field image (red arrows, left image) are not visible in the fluorescent image on the right. Red blood cells are excluded from cell counts.
Because platelets and mature mammalian red blood cells don't contain nuclei, only mononuclear cells produce a fluorescent signal. Fluorescent-positive live and dead nucleated cells are easily counted and red blood cells are excluded from cell counts. There is no need to lyse red blood cells, saving time and eliminating an extra sample preparation step. This also allows technicians to use a single viability method for analysis of both fresh and processed blood, bone marrow, and digested tissue samples.
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In addition to dual-fluorescence viability, the Vision includes a default assay for cell concentration and % viability for cultured cells stained with trypan blue.
Trypan blue viability is a dye exclusion method that utilizes membrane integrity to identify dead cells. The dye is unable to penetrate healthy cells, so they remain unstained. Dead cells have a compromised cell membrane that is permeable to the trypan blue dye. Dead cells are stained blue and display as dark cells in the Cellometer software with bright field imaging.
Other bright field stains, such as Methylene Blue and Erythrosine B, can also be analyzed using Cellometer cell counters.
Live cells are circled in green. Dead cells are circled in red.
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Primary hepatocytes (both fresh and cryo-preserved) are routinely used to test potential drug candidates for toxicity. Seeding with identical numbers of viable cells is important for accurate toxicity experiments. Due to their variable morphology and tendency to clump, hepatocytes can be difficult to count manually or with other automated cell counters. The Cellometer Vision features specialized algorithms for accurate counting of hepatocytes. Dual-staining with AO/PI enables counting of live and dead hepatocytes for automated viability determination.
Bright field image (left) shows the variable morphology of primary hepatocytes. Dual fluorescence image (right) shows counted live cells (circled in green) and counted dead cells (circled in red).
Hepatocytes are quite fragile. Because the Vision images cells directly from the counting chamber, the shear stress present in flow-based systems (where cells travel through the instrument) is eliminated.
Because hepatocytes lose viability over time, it is important to complete the analysis very quickly. Staining with the Cellometer AO/PI viability reagent and loading of the counting chamber takes just 1 minute. Analysis with the Cellometer Vision is completed in less than 60 seconds.
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The Cellometer Vision (10x) Image Cytometry System is optimized for simple, 1-step determination of yeast concentration and viability in messy samples. Fluorescent nuclear staining dyes are used to stain live and dead nucleated cells. Learn more about the AO/PI dual-fluorescence method.
The Cellometer Yeast Viability Kit was specifically developed for dual-fluorescence analysis of yeast in samples containing corn mash. Reagents for viability of yeast in samples containing corn stover, sugar cane, and biomass are also available. Contact Nexcelom technical support for more information.
Bright field and fluorescent images of yeast from corn mash. Debris visible in the bright field image (above left) is excluded in the dual-fluorescence image (above right). Only live (circled in green) and dead (circled in red) yeast are counted.
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Including the NCI-60 human cancer cell lines developed by the National Cancer Institute.
The MCF-7 breast cancer cell line can be very clumpy. The Cellometer pattern-recognition software identifies and counts individual cells within these cell clumps for accurate analysis (shown at right).
For clumpy cells with poorly-defined edges, the Vision offers fluorescent counting. Cells can be stained with a nuclear dye, such as acridine orange (AO), then imaged and counted in the fluorescent channel.
The Cellometer cell roundness setting can be adjusted for recognition and counting of irregular-shaped cells, such as RD cells, activated T-cells, and hepatocytes.
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Nexcelom has designed specialized (PD-300) counting chambers to accommodate large cells, including adipocytes. The controlled chamber depth eliminates potential problems caused by the buoyancy of adipocytes. Imaging directly from the counting chamber eliminates the shear stress present in flow-based systems where cells travel through the instrument, making accurate analysis of fragile adipocytes possible.
Optimized bright field counting parameters or fluorescent stains can be used to exclude free lipid droplets (shown at right) from counting results. Optimized settings for adipocyte analysis are saved as a unique cell type
Bright field image of adipocytes
Bright field counted image of adipocytes
Dual fluorescence live / dead counted image of adipocytes
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Because Cellometer recognizes cells based on size, brightness, and morphology, cellular debris is easily and accurately excluded from counting results.
The Cellometer Vision automatically generates a cell size histogram based on cell diameter.
Adipocyte cell size histogram showing variation in cell diameter from 44 to 202 microns.
Minimum and maximum cell diameter settings in the Cellometer software can be optimized to count specific cells in a sample. The example below demonstrates counting of mature dendritic cells cultured from PBMCs based on cell diameter.
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Bright field and fluorescent counted images of macrophages stained with Calcein-AM for detection of metabolically-active cells. Debris and dead cells in the bright field image (left) are not visible in the fluorescent image (right).
View Bright Field Images to check cell morphology.
View Fluorescent Counted Images to:Automatically Save raw images and data to a secure network location.
Export Data to Excel.
Capture Colorized Images with screen-capture software for use in publications and presentations.
Print images and custom data reports directly from Cellometer software and FCS Express.
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Optics Module | Fluorophores | Nucleic Acid Stains | Fluorescent Proteins |
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VB-450-302 Ex: 375 nm Em: 450 nm |
AlexaFluor® 350 | DAPI Hoechst 33342 Hoechst 33258 |
BFP CFP |
VB-535-402 Ex: 475 nm Em: 535 nm |
Calcein FITC AlexaFluor® 488 |
AO (acridine orange, +DNA) SYTO®9, SYTO®13 |
GFP YFP |
VB-595-502 Ex: 525 nm Em: 595 nm |
AlexaFluor® 546 AlexaFluor® 555, Cy3® PE (R-phycoerythrin) Rhodamine B |
PI (propidium iodide) EB (ethidium bromide) SYTOX® Orange |
57.5 |
VB-660-502 Ex: 540 nm Em: 660 nm |
AlexaFluor® 647 7-AAD Nile Red |
PI (propidium iodide) EB (ethidium bromide) AO (acridine orange, +RNA) |
Ds Red RFP TdTomato |
VB-695-602 Ex: 630 nm Em: 695 nm |
AlexaFluor® 647, Cy5® APC (allophycocyanin) |
SYTOX® Red | 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.
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Cellometer Disposable Counting Chambers consist of two independent enclosed chambers with a precisely controlled height. Cell suspension of 20 microliters is loaded into the chamber using a standard single channel pipette.
The chamber is inserted into the Cellometer cell counter and the cells are imaged directly from the chamber. This simple sample loading and analysis method is ideal for fragile cells.
The disposable Cellometer Cell Counting Chambers offer several key advantages:
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On-site demonstrations allow users to evaluate a critical cell type or cell-based assay prior to making a purchase.
Technical seminars are a great way to introduce Cellometer cell viability and cell-based assays to multiple users within a lab or organization.
Free on-site installation and training are available to all Vision and Vision CBA customers.
Web-based support sessions are available to all Cellometer customers at no cost. Nexcelom technical support specialists can assist with creation of new cell types, optimization of counting parameters, and analysis of cell-based assays.
Comprehensive Protocols for a growing menu of cell-based assays assist with sample preparation, imaging, and data analysis. Detailed, step-by-step instructions with screen shots enable new users to run Cellometer kits for cell cycle, apoptosis, cell viability, and other assays.
On-Line Help can be accessed directly from the Vision and Vision CBA Instruments. Users can:
Application or Cell-Based Assay |
Vision
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Vision CBA
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Live/Dead Cell Concentration & Trypan Blue Viability (clean cell samples) |
X
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X
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Live/Dead Nucleated Cell Concentration & Dual-Fluorescence Viability (samples with and without debris, RBC, platelets; no lysing) |
X
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X
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Small Cells: Yeast, Algae, Platelets |
10x
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10x
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Hepatocyte and Adipocyte Analysis |
X
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X
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Cell Cycle: PI |
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X
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Apoptosis: Annexin V-FITC/PI, Caspases, Mitochondrial Membrane Potential |
X
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X
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Autophagy |
X
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X
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GFP, RFP, YFP, and other fluorescent proteins* |
X
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X
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Surface Markers: FITC*, PE*, APC* |
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X
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Proliferation: CFSE* |
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X
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Apoptosis: Chromatin Condensation* |
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X
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X = numbers only X = with population plots and histograms
*Enhanced sensitivity of Vision CBA is strongly recommended for this application
*Enhanced sensitivity of Vision CBA may be required if signal is weak
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User-changeable fluorescence optics modules compatible with the Cellometer Vision and Vision CBA Systems
Catalog # | Description | Compatible Dyes / Fluorophores* | Unit |
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VB-450-302 | Cellometer Vision Optical Module Excitation / Emission: 375nm/450nm |
DAPI, Hoechst 33342 and 33258, BFP, CFP | each |
VB-535-402 | Cellometer Vision Optical Module Excitation / Emission: 475nm/535nm |
Acridine Orange (AO) (+DNA), Fluorescein (FITC), SYTO®9, SYTO®13, Calcein, Fluorescein (FITC), AlexaFluor®488, GFP, YFP | each |
VB-595-502 | Cellometer Vision Optical Module Excitation / Emission: 525nm/595nm |
Propidium Iodide (PI),Ethidium Bromide (EB), SYTOX® Orange, AlexaFluor®546, AlexaFluor®555, Cy3®, R-phycoerythrin (PE), Rhodamine B, Ds Red, RFP, TdTomato | each |
VB-660-502 | Cellometer Vision Optical Module Excitation / Emission: 540nm/660nm |
Propidium Iodide (PI),Ethidium Bromide (EB), Acridine Orange (AO) (+RNA), AlexaFluor®647, 7-AAD, Nile Red | each |
VB-695-602 | Cellometer Vision Optical Module Excitation / Emission: 630nm/695nm |
Allophycocyanin (APC), AlexaFluor®647, Cy5®, SYTOX® Red, Crimson | each |
Software upgrades available for the Cellometer Vision CBA Instrument
Catalog # | Description | Size | Unit |
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VSL-01 | Cellometer Vision Software License for GMP/GLP Support | Single-user copy | 1 license |
FCS-SW4RP-10 | Additional single-user copy of FCS Express 4 Flow, RUO Professional Software | Single-user copy | 1 license |
Installation Qualification (IQ) and Operational Qualification (OQ) package for the Vision instrument.
Service | Description | Size | Unit |
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Cellometer Vision-IQOQ | Counting chambers, reference bead solution, and IQ/OQ validation protocol for the Cellometer Vision | 100 counts | 1 Set |
Trained Nexcelom Applications Specialists are available for on-site preventative maintenance, IQ validation, and OQ validation.
Service | Description |
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Preventative Maintenance |
Available for all Cellometer Systems Please inquire for scheduling and pricing. |
IQ Validation* |
Available for Auto T4, Auto X4, and Vision Systems Please inquire for scheduling and pricing. |
OQ Validation* |
Available for Auto T4, Auto X4, and Vision Systems Please inquire for scheduling and pricing. |
Extended Warranty |
Available for all Cellometer Systems currently under a valid warranty. Please inquire for pricing. |
Models and Specifications
Cellometer Vision CBA | |||||||||||||||||||||||||||||||||
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Includes: |
· Cellometer Vision HSL Instrument · PC Laptop · Two Standard Fluorescence Optics Modules · Cellometer Vision CBA Software (2 copies: 1 for instrument, 1 for remote work station) · USB 2.0 Connection Cable · Power Supply · Phone/online applications support during set-up · 75 slides |
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Optional Components /
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10x Magnification (5x magnification is standard) | ||||||||||||||||||||||||||||||||
Imaging Performance: |
Cell Size: Conc. Range: |
Vision 5x 5 - 300* microns 105 - 107 cells/ml |
Vision 10x 0.5 - 25 microns 105 - 107 cells/ml |
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*Cellometer CHT4-PD300 Slides are required for cells > 80 microns in diameter | |||||||||||||||||||||||||||||||||
Instrument
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Weight: 25 lbs. (11 kg) Dimensions: Width: 6" (15cm) Depth: 8.5" (22cm) Height: 14" (36cm) Voltage: 100-240V AC 50-60 Hz |
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Available Fluorescence Optics Modules: |
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