The Cellometer Auto 2000 cell viability counter from Nexcelom Bioscience is a touchscreen, fluorescent automated instrument specifically designed for primary samples.

 

Simple and User-Friendly Procedure

Cellometer Auto 2000

 

  • Pipette 20 µL
  • Insert slide
  • Choose assay and click count
  • View results in 30 seconds

Simple, Automated Cell Counting in 30 Seconds

The Cellometer Auto 2000 cell viability counter uses dual-fluorescence imaging and bright-field imaging to rapidly and precisely detect and count individual cells. Count, diameter, concentration, and % viability of cells are calculated and reported automatically.

Load sample, view image, count cells, and obtain results in less than 30 seconds.

The Auto 2000 enables users to:

  • Enhance consistency
  • Increase precision
  • Increase throughput
  • Avoid judgment errors, interference from red blood cells, miscounts, and user-to-user variability
  • Count complex cells (both irregular-shaped and clumpy)
  • Guarantee that all data is properly captured

Primary Cell Analysis: PBMCs, Stem Cells and More

The Cellometer Auto 2000 has been particularly optimized to analyze primary cells from bone marrow, cord blood, peripheral blood, and other complex samples for use in various research areas, such as:

  • Suspensions of tumor cells for oncology
  • Stem cells for cellular therapy
  • Splenocytes for development of vaccines
  • PBMCs for immunology
  • Nucleated cells for transplantation

Both live and dead nucleated cells can be stained with dual-color fluorescence. This produces precise viability results even in the presence of red blood cells, platelets, and debris. Precise analysis of both “clean” and “messy” samples allows the Cellometer Auto 2000 to assess samples at many different points throughout sample processing, right from initial collection through to separation and cryopreservation.

The Cellometer Auto 2000 includes one-touch assays for inspecting a variety of primary samples, such as:

Nucleated Cells from Whole Blood

Cellometer Auto 2000

 

Stem Cells

Cellometer Auto 2000

 

PBMCs/Splenocytes Following Separation

Cellometer Auto 2000

 

Tumor and Tissue Cell Suspension

Cellometer Auto 2000

 

No Interference from Red Blood Cells, Platelets, or Debris

The dual-fluorescence AO/PI technique uses nuclear staining dyes that adhere to nucleic acids in the cell nucleus. Since a majority of the mature mammalian red blood cells lack nuclei, only live mononuclear cells and dead mononuclear cells create a fluorescent signal. This means, red blood cells do not have to be lysed, which saves time and prevents the additional sample preparation step. Debris, platelets, and red blood cells are not counted in the fluorescent channels.

The Advantage of Fluorescent Counting For Primary Cells

The following images reveal the benefit of fluorescent counting for primary cells. The bright-field image demonstrates the combination of platelets, red blood cells, and nucleated cells present in the sample. Only the live nucleated cells and dead nucleated cells are observed and counted in the red and green fluorescent channels.

Sample Measurement Total nucleated All RBC % RBC n
PBMC+RBC Mean

 

CV

1.26E+07

 

6.2%

1.39E+07

 

8.8%

1.23E+06 8.9% 10
PBMC+1/2RBC Mean

 

CV

1.21E+07

 

4.8%

1.27E+07

 

5.4%

5.82E+05 4.6% 10
PBMC+1/4RBC Mean

 

CV

1.22E+07

 

7.9%

1.24E+07

 

7.3%

2.27E+05 1.8% 10
 

Fresh human peripheral blood mononuclear cells (PBMCs) were spiked with different amounts of red blood cells. All cells (both RBC and nucleated) were counted in the bright-field channel. Subsequently, nucleated cells were counted in the green fluorescent channel. Different amounts of red blood cells (1.8%, 4.6%, and 8.9%) did not have any impact on the nucleated cell count.

Cellometer Auto 2000

 

The bright-field image illustrated below indicates many red blood cells. While these red blood cells are not seen in the fluorescent image shown below, cells stained with nuclear staining dye have been identified.

Analysis of Clumpy and Irregular-Shaped Cells

Including NCI-60 and Clumpy MCF-7 Cells

NCI-60 is a series of 59 human cancer cell lines, which were originally 60 human cancer cell lines and produced by the National Cancer Institute for screening purposes.

  • Of the NCI-60 cell lines, 57% are clumpy, exhibit a huge difference in cell size or shape, or contain debris
  • The entire 59 NCI-60 cell lines have been effectively verified on the Cellometer Auto 2000 cell counter

Clumpy Cells

The MCF-7 breast cancer cell line can be quite clumpy. The Cellometer pattern-recognition software detects and counts each cell inside these cell clumps for precise analysis as illustrated in the image below.

Cellometer Auto 2000

 

Irregular-Shaped Cells

Users can adjust the cell roundness setting of the Cellometer to identify and count irregular-shaped cells, like activated T-cells and RD cells.

Cellometer Auto 2000

 

Cellometer cell counters are used by all 40 NCI Comprehensive Cancer Centers.

User-Friendly Touch Screen

The easy-to-use touch screen shows pre-optimized assays for normal types of cells. User-defined protocols can be easily produced and saved to the menu. The pre-optimized and simple assays make it easy to operate the instrument and streamline training for novice users.

Cellometer Auto 2000

 

By simply clicking a button, scientists can choose a fresh assay with saved counting parameters. This makes the instrument perfect for testing a wide range of cell types in laboratories. Using auto-save options, users can rapidly test a range of samples.

Live/Dead Nucleated Cell Counts Using Dual-Fluorescence

Green Fluorescent Live Cell Image

Cellometer Auto 2000

 

Red Fluorescent Dead Cell Image

Cellometer Auto 2000

 

Why Dual-Fluorescence?

Since bright-field cell counting does not differentiate between nucleated cells and non-nucleated cells and Trypan blue staining cannot be as easily detected as fluorescent staining, dual-color fluorescence is highly advised for precise viability analysis of primary cells.

The Cellometer Auto 2000 Cell Viability Counter is fitted with regular assays for dual-fluorescence analysis of AO/PI-stained primary cells.

The AO/PI Method

Acridine orange (AO) is a nuclear staining (binding of nucleic acid) dye that is permeable to both live cells and dead cells. It creates green fluorescence by staining all nucleated cells.

Propidium iodide (PI) can only penetrate dead cells with substandard membranes, and generates red fluorescence by staining all the dead nucleated cells. Both PI- and AO-stained cells fluoresce red because of quenching, and therefore, all the dead nucleated cells fluoresce red, while all the live nucleated cells fluoresce green.

Cell Images for Data Verification

No Two Cells are the Same

With the Cellometer Auto 2000 cell counter, the morphology of cells can be instantly visualized on-screen in the bright-field image. Counted cells are indicated on-screen to further verify that cells present in the sample are being imaged and studied correctly. Bright-field counted images can be visualized for Trypan blue viability and basic cell counting.

Fluorescent counted images denoting counted live nucleated cells and dead nucleated cells can be visualized for dual-fluorescence primary cell viability assays.

Users can confirm whether:

  • Cells have been counted properly, depending on their shape and size
  • Platelets, red blood cells, and debris have been excluded from results
  • Cells inside the clumps have been counted separately

Cellometer Auto 2000

 

Cellometer Auto 2000

 

The bright-field image demonstrates that smaller debris is not being counted, while individual cells within pairs are being counted. Live counted cells in the integrated fluorescent counted image are circled in green, and dead counted cells in the same image are circled in red.

  • Users can archive and export the images of the cells for use in presentations and publications
  • Users can re-count the saved images using user-defined or default analysis settings

Cell Size Analysis and Size-Based Counting

Cellometer Auto 2000

 

The Auto 2000 Cell Counter automatically creates a cell size histogram depending on the diameter of cells.

Since the Cellometer Auto 2000 cell counter creates measurements of individual cell size, various samples can be superimposed on a single histogram, thus making it possible to analyze the changes that occur in the diameter of cells over time.

Cellometer Auto 2000

 

In experiments where SF21 or SF9 cells are used, saved images can be visualized and re-counted to assess the changes in cell morphology over time. Settings of maximum and minimum cell diameter can be improved to count specific cells present in a sample.

Cellometer Auto 2000

 

Such an example shows the counting of mature dendritic cells that were cultured from PBMCs based on the diameter of cells.

10x Faster than Manual Counting

It takes about 5 minutes to count 1 x 106 cells with a manual hemacytometer. At times, it takes twice the time to count live cells and dead cells. The Cellometer Auto 2000 Cell Viability Counter estimates the cell count and concentration for both dead and live cells and also calculates % viability in only 30 seconds.

Cellometer Auto 2000

 

Data Accuracy and Consistency can be Improved

  • Judgment errors can be prevented
  • Wash steps can be eliminated
  • Recording and calculation errors can be prevented
  • RBC interference can be eliminated
  • Counting time can be reduced, allowing users to run more experiments

Cellometer Precision

The Cellometer Auto 2000 cell counter provides exceptional reproducibility, with a % coefficient of variation (CV) of less than 10% for fluorescent concentration as well as viability measurements.

The following data is based on four preparations of Jurkat cells stained with a fluorescent nuclear-staining dye called propidium iodide.

.

Sample N Value Average Live Cell Concentration % Viability CV of Concentration CV of Viability
A 4 4.20E+06 91.1 10% 2%
B 4 1.06E+06 22.7 7% 1%
C 4 3.27E+06 57.5 7% 7%
 

Imaging/Counting Chambers: No Washing or Contamination

Cellometer Auto 2000

 

The Cellometer Disposable Imaging Chambers contain a pair of separately enclosed chambers with an accurately controlled height. A 20 µL cell suspension is placed into the chamber using a regular single-channel pipette. This chamber, in turn, is embedded into the Cellometer cell counter and the cells are finally imaged.

This method of sample loading and analysis is simple and suitable for fragile cells.

The disposable Cellometer cell counting chambers provides many major benefits:

  • Risk of cross-contamination is eliminated
  • The washing step is eliminated, which saves time
  • A controlled volume of samples is achieved
  • Decreased biohazard risk to users
  • The most affordable automated counting consumables
  • Large-depth chambers for huge cells

Auto-Save Data and Cell Images

The Cellometer Auto 2000 cell counter enables users to produce specific folders in which images and data reports can be saved automatically. With this auto-save feature, users can ensure that all the data is precisely recorded in the right place. This feature also allows users to test a range of samples quite rapidly.

Cellometer Auto 2000

 

Users can also configure the Cellometer Auto 2000 cell counter to request a new sample name for every sample that is imaged and counted, or alternatively, they can apply auto-numbering to each sample in a series to further decrease the analysis time. In addition, users can apply a time stamp to individual data files.

Cellometer Auto 2000

 

Dedicated On-line and On-Site Applications Support

At Nexcelom Bioscience, experienced technical support specialists are available from 8:30 am to 5:00 pm EST via phone and on-line support and can help customers with:

  • Training of novice users
  • Troubleshooting
  • Generation of new cell types
  • Optimization of counting parameters
  • Installation of the Cellometer Auto 2000 cell viability counter

By using the help button provided at the bottom right of the Cellometer Auto 2000 software screen, users can get quick access to:

  • Online training videos and tutorials
  • Features and instructions of the software
  • Submission of a Support Ticket

All applications specialists at Nexcelom Bioscience are fully focused on cell-based assays and image-based concentration and viability of cells using Cellometer Image Cytometry.

Nexcelom field-based applications specialists are also available for:

  • Troubleshooting
  • Training
  • Technical seminars
  • On-site demonstrations