Nexcelom Bioscience

978-327-5340

Applications for Cellometer Auto 2000 Cell Viability Counter

Adipocytes

Adipocytes

Automatically measure cell size of freshly isolated adipocytes and plot size histogram. DNA staining fluorescence dyes are used to identify cells from lipid droplets. »

Adoptive Cell Transfer Therapy

Adoptive Cell Transfer Therapy

Use Cellometer to perform cell based assays and measure cell size, viability and concentration of cell lines and primary samples used in adoptive cell therapy research. »

Trypan Blue and AO/PI

Cell Viability Measurement Using Trypan Blue or AO/PI

When should you use trypan blue and when should you use acridine orange/propidium iodide to measure cell viability? »

Cell Viability and Necrosis

Cell Viability and Necrosis

Cell viability is performed using various fluorescent membrane exclusion dyes, such as PI, EB, 7AAD, and others. This assay is performed by enumerating cells in captured bright-field and fluorescent images. And Necrotic cells are detected using propidium iodide. »

Blood Samples

TNC Concentration & Viability for Clinical (Blood) Samples

Analyze fresh and processed blood and bone marrow samples without lysing: no interference from RBCs. »

Cell Size Assay

Cell Size Assay

Performing cell size measurement assay and using cell size to count cells within preset cell size parameters. For adipocytes, stem cells, Sf9 cells, dendritic cells, and others. »

Cancer Cell Lines

NCI-60 Cancer Cell Lines

Automatically measure live cell concentration and viability of cancer cell lines used in oncology research and most of all biology research. »

GFP Transfection Efficiency

Quantitative Measurement of GFP Transfection

Rapidly identify fluorescence positive cells from a sample, analyze individual cell fluorescence intensity, calculate cell concentration, size and determine the GFP transfection automatically. »

Immunology

Immunology Research

Automatically quantify cell viability and concentration for a variety of immunologically relevant samples such as: bone marrow, cord blood, slpenocytes, lymphocytes, isolated mononuclear cells, tumor digests, murine samples, and others. »

Insect Cells

Insect Cells

Automatically measure live cell concentration, viability for baculovirus infected insect cells. Cell size histogram live cell concentration and viability are generated within less than 60 seconds using 20 µl sample. »

PBMC

Peripheral Blood Mononuclear Cells (PBMC)

Automatically measure live cell concentration and viability without lysing red blood cells for consistent results from patient samples. Other cells include splenocytes and bone marrow. »

Platelets

Platelets

Automatically measure mouse and human platelet concentration using brightfield only method. »

Stem Cells

Stem Cells

Automatically measure live cell concentration and viability for stem cells, such as human and mouse ES cells, mesenchymal, cardiac, induced pluripotent stem cells. Multiple viability stains, such as trypan blue, propidium iodide are used to identify dead cells. For samples with a lot of debris, dual fluorescent nuclear stains are used for live and dead cells. »

WBSc in Whole Blood

WBCs in Whole Blood

Automatically measure nucleated cell concentration without lysing red blood cells using nuclear staining dyes (AO), for human and mouse blood. »

Stem Cells, Primary Cells, Cell Line

Stem Cells, Primary Cells, Cell Line
Figure 2: Chart of the % viable cells at 3 different live/dead mixture ratios. N=4 per measurement. The error bars correspond to the standard deviation for each measurement.
Sample
N Value
Average Live Cell Concentration
% Viability
CV of Concentration
CV of Viability
A
4
4.10E+06
78
1%
2%
B
4
0.0025E+06
0.6
N/A
N/A
C
4
2.48E+06
36
4%
7%
Figure 1: Table of results for AOPI viability using the Cellometer Auto 2000.
The results indicate the accuracy of the Cellometer Auto 2000 instrument in assessing the viability of Jurkat cells using AOPI for cell viability. Four measurements were performed for each sample. The viability average was calculated and plotted. The results show the reliability and accuracy of the Cellometer Auto 2000 in measuring cell concentration and viability of mammalian cells.

Cell Line Viability using Propidium Iodide Only

Cell Line Viability using Propidium Iodide Only
Figure 2: Chart of the % viable cells at 3 different live/dead mixture ratios. N=4 per measurement. The error bars correspond to the standard deviation for each measurement.
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%
Figure 1: Table of results for cell viability using PI only.
The results indicate the accuracy of the Cellometer Auto 2000 instrument in assessing the viability of Jurkat cells using PI for cell viability. Four measurements were performed for each sample. The viability average was calculated and plotted. The results show the reliability and accuracy of the Cellometer Auto 2000 in measuring cell concentration and viability of mammalian cells.

Cell Line Total Cell Concentration

Cell Line Total Cell Concenration
Figure 2: Jurkat cell concentrations at various sample dilution factors. N=4. Error bars correspond to the Standard Deviations for cell concentration.
Sample
Dilution Factor
Average Cell Concentration
CV of Concentration
Standard Deviation
N Value
A
1
5.80E+06
1%
4.8E+04
4
B
2
3.39E+06
2%
8.12E+04
4
C
4
1.63E+06
6%
9.54E+04
4
D
8
8.27E+05
6%
5.33E+04
4
Figure 1: Table of results for total cell concentration for the Cellometer Auto 2000 instrument.
The results indicate the accuracy of the Cellometer Auto 2000 instrument cell concentration. Serial dilutions (2-, 4-, and 8-fold) were prepared using PBS and Jurkat cells. Four measurements were performed for each sample and the averages were calculated and plotted. A linear correlation between the concentration measurements and 1/dilution factor was observed. The results show the reliability and accuracy of the Cellometer Auto 2000 in measuring cell concentration.

Low Concentration of Cells using Acridine Orange

Low Concentration of Cells using Acridine Orange
Figure 2: Graph of Concentration for 4 different measurements using the Low Concentration of Cells Assay and AO on the Cellometer Auto 2000.
Low Concentration AO
F1 Concentration
Sample 1
5.91E+04
Sample 2
6.40E+04
Sample 3
6.58E+04
Sample 4
6.90E+04
Average
6.45E+04
Stdev
4.14E+03
CV
6%
Figure 1: Table of results from the Low Concentration of Cells assay.
The results indicate the accuracy of the Cellometer Auto 2000 instrument when counting Jurkat cells at very low concentrations (between 5-7E+04 cells/ml). AO was used to measure the concentration of cells in the F1 channel. Results from 4 measurements were plotted. The CV values are very close, which indicates the consistency of the measurements. The results show the reliability and accuracy of the Cellometer Auto 2000 in measuring at low cell concentration.

Performance of the Cellometer Auto 2000 Cell Viability Counter

Consistency and Accuracy Comparison to Hemacytometer

Jurkat

N = 20 Hemacytometer Auto 2000
Average 1.03E+06 1.01E+06
STDEV 6.60E+04 8.87E+04
%CV 6.4% 8.8%

5 µm beads

N = 20 Hemacytometer Auto 2000
Average 1.07E+06 1.03E+06
STDEV 5.90E+04 4.30E+04
%CV 5.5% 4.2%