Nexcelom Bioscience
Cellometer Vision Image Cytometry

The Vision CBA is a PC-based image cytometer for automated image capture and analysis. The system performs 5x or 10x bright field and dual channel fluorescent cell counting, viability, intensity, and population analysis with flow-like data output and interchangeable fluorescent modules. Image analysis software handles clumpy and irregularly shaped cells and omits debris for accurate measurements using a variety of cell types, including cell lines, PBMCs, hepatocytes, adipocytes, yeast, and algae. Combining with the FCS Express software, cell-based assays such as Cell Cycle, Apoptosis, Autophagy, Transfection Efficiency, Surface Marker Immunophenotyping can be easily performed.

Simple, 20µl Cell-Based Assays

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:

  • Validated assay kits and reagents
  • Simple staining and analysis procedures
  • Default settings for automated data generation
  • No washing. No clogging. No daily calibration.
Cell Cycle
Annexin V data plot

Growing Menu of Assays

Fluorescent Cell-Based Assays
Vision CBA
Apoptosis (Annexin V-FITC / PI)
Watch Video
Apoptosis (Caspase Activity)
Autophagy (CytoID-green)
Cell Proliferation (CFSE)
Cell Cycle (PI)
Watch Video
GFP, RFP, YFP Transfection
Mitochondrial Potential / Early Cell Death (JC-1)
Watch Video
Multi-Drug Resistance (ABC Transporter)
Surface Marker Analysis (PE, APC, FITC)
Viability (AO/PI, Trypan blue)
Vitality (Calcein-AM / PI)

Flow-Like Data Output

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.

  • Cell Images
  • Data Tables
  • Dot Plots
  • Histograms

Data layouts can be modified to accommodate multi-sample analysis and create customized analysis and / or quality control reports.

Back to top ↑

Nucleated Cell Counting and Viability in <60 Seconds

Count Live Nucleated Cells

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.

Count Dead Nucleated Cells

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.

Live cells stained with AO

Live cells stained with AO

Dead cells stained with PI

Dead cells stained with PI
Red blood cells are excluded from cell counts.

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.

No Interference from RBCs, Platelets, or Debris

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.

Back to top ↑

Trypan Blue Viability

trypan blue viability

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.

Counted Trypan Blue Bright Field Image

Counted Erythrosin B Bright Field Image

Counted Trypan Blue  Bright Field Image  Counted Erythrosin B Bright Field Image

Live cells are circled in green. Dead cells are circled in red.

Fast

Within 30 seconds, the Vision instrument reports:

  • Cell concentration (total, live, and dead)
  • Cell number (total, live, and dead)
  • Mean cell diameter (total, live, and dead)
  • Percent viability

Back to top ↑

Hepatocyte Analysis

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.

Hepatocyte Analysis

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).

Simple Imaging Method Ideal for Fragile Cells

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.

Fast Analysis for Accurate Viability Determination

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.

Back to top ↑

Yeast Viability for BioFuel Production with Vision 10x

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.

Yeast Viability for BioFuel Production

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.

Back to top ↑

Analysis of Clumpy and Irregular-Shaped Cells

Clumpy Cells

Including the NCI-60 human cancer cell lines developed by the National Cancer Institute.

Clumpy Cells

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.

Irregular-shaped Cells

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.

  • More than 1,600 different cell lines have been successfully counted using Cellometer Systems
count irregular-shaped cells

Back to top ↑

Adipocyte Analysis

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

Exclude free lipid droplets
Bright field image of adipocytes

Bright field image of adipocytes

Bright field counted image of adipocytes

Bright field counted image of adipocytes

Dual fluorescence live / dead counted image of adipocytes

Dual fluorescence live / dead counted image of adipocytes

Back to top ↑

Exclusion of Debris & Size-Based Counting

Because Cellometer recognizes cells based on size, brightness, and morphology, cellular debris is easily and accurately excluded from counting results.

  • Cell size parameters can be modified to optimize exclusion of debris from results and enhance the accuracy of counting for a wide range of cell sizes.
  • A fluorescent nuclear dye, such as acridine orange, can be used to more easily identify nucleated cells in samples containing debris.
Counted cell image excludes debris

Cell Size Histogram

The Cellometer Vision automatically generates a cell size histogram based on cell diameter.

Cell Size Histogram

Adipocyte cell size histogram showing variation in cell diameter from 44 to 202 microns.

Size-Based Counting

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.

Cell Size Dendritic Cells

Back to top ↑

View, Print, and Save Cell Images and Data Tables

Confirm counted cells

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:
  • confirm exclusion of debris
  • confirm exclusion of unwanted cell types
  • optimize counting of irregular-shaped cells
  • confirm counting of individual cells within clumps
Save raw images and data

Automatically Save raw images and data to a secure network location.

Export data to Excel

Export Data to Excel.

Capture colorized images

Capture Colorized Images with screen-capture software for use in publications and presentations.

Print data

Print images and custom data reports directly from Cellometer software and FCS Express.

Back to top ↑

User-Changeable Fluorescence Optics Modules

The Vision CBA Image Cytometry System features two user-changeable fluorescence optics modules. The robust modules are easily removed and replaced. Available modules cover a spectrum from UV to far-red, enabling analysis of a wide range of fluorescent proteins, dyes, and fluorophores.
User-changeable optics modules
DAPI, Sytox Green, EB, PI 7-AAD, Sytox-Red cell images
Optics Module Fluorophores Nucleic Acid Stains Fluorescent Proteins
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
RFP
Ds Red
TdTomato
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)
 
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.

Back to top ↑

Counting Chambers: No Washing or Contamination

Cell Counting Chamber

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:

  • Time savings – no washing
  • No risk of cross-contamination
  • Reduced biohazard risk to users
  • Controlled sample volume
  • Controlled chamber depth
  • Direct imaging from chamber
  • Most affordable automated counting consumables

Back to top ↑

Dedicated On-line and On-site Applications Support

Nexcelom Applications Specialists focus exclusively on cell counting, cell viability, and cell-based assays.
On-site demonstrations

On-site demonstrations allow users to evaluate a critical cell type or cell-based assay prior to making a purchase.

Technical seminars

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

Free on-site installation and training are available to all Vision and Vision CBA customers.

Web-based support sessions

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

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.

Help

On-Line Help can be accessed directly from the Vision and Vision CBA Instruments. Users can:

  • View training videos
  • View frequently asked questions
  • Download instructions and software updates
  • Submit a Support Ticket directly to Nexcelom

Cellometer Vision vs. Vision CBA: Which System is Right for Me?

The Cellometer Vision and Vision CBA are specifically optimized for concentration and viability analysis of complex cell samples and unique sample types. Unlike the standard Cellometer Vision, the Vision CBA features an additional light source (HSL Module) for enhanced sensitivity and FCS Express 4 Flow software for advanced population analysis based on fluorescence intensity. Assay data is displayed in a table and also as a scatter plot or histogram for simple identification and measurement of unique cell populations. The table below compares Vision and Vision CBA capabilities for a number of popular cell types and cell-based assays.
Application or Cell-Based Assay
Vision
Vision CBA
Live/Dead Cell Concentration & Trypan Blue Viability (clean cell samples)
X
X
Live/Dead Nucleated Cell Concentration & Dual-Fluorescence Viability (samples with and without debris, RBC, platelets; no lysing)
X
X
Small Cells: Yeast, Algae, Platelets
10x
10x
Hepatocyte and Adipocyte Analysis
X
X
Cell Cycle: PI
X
Apoptosis: Annexin V-FITC/PI, Caspases, Mitochondrial Membrane Potential
X
X
Autophagy
X
X
GFP, RFP, YFP, and other fluorescent proteins*
X
X
Surface Markers: FITC*, PE*, APC*
X
Proliferation: CFSE*
X
Apoptosis: Chromatin Condensation*
X

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

Back to top ↑

Cellometer Vision Accessories

Cellometer Vision / Vision CBA Fluorescence Optics Modules

User-changeable fluorescence optics modules compatible with the Cellometer Vision and Vision CBA Systems

Catalog # Description Compatible Dyes / Fluorophores* Unit
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
*An incomplete listing of popular fluorophores, dyes, and fluorescent proteins that are compatible. Customized optical modules in the UV to far-red range are available. E-mail support@nexcelom.com or call 978-327-5340 for more information.

Software

Software upgrades available for the Cellometer Vision CBA Instrument

Catalog # Description Size Unit
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

IQ / OQ Validation Package

Installation Qualification (IQ) and Operational Qualification (OQ) package for the Vision instrument.

Service Description Size Unit
Cellometer Vision-IQOQ Counting chambers, reference bead solution, and IQ/OQ validation protocol for the Cellometer Vision 100 counts 1 Set

On-Site Service, Maintenance, and Instrument Warranties

Trained Nexcelom Applications Specialists are available for on-site preventative maintenance, IQ validation, and OQ validation.

Service Description
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.
*IQ/OQ Validation Package must be purchased separately

Back to top ↑

Cellometer Vision CBA Resources