Nexcelom Bioscience

Direct Count of White Blood Cells from Peripheral Blood Sample
without Lysing Red Blood Cells


Compatible with Cellometer:

Cellometer Vision Image Cytometer
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Brightfield image of whole blood.
Brightfield image of whole blood.
WBCs are not visible amongst RBCs

Fluorescent image showing AO stained white blood cells
Fluorescent image showing AO
stained WBCs. RBCs are not visible.

Cellometer Vision incorporates image based cell counting
and fluorescence detection in a compact and easy-to-use
instrument. With fluorescence detection capabilities,
Cellometer Vision is an ideal solution for many complex cell population characterization assays such as rapidly counting white-blood-cells in whole blood.

As one of the most important cellular components in peripheral blood, the concentration of white blood cells is routinely used in researching various diseases, such as bacteria/virus infections, leukemia, or leucopenia. However, conventional methods to count white blood cells include a time consuming and tedious RBC lysis process.

Vision can be used to simplify traditional methods by mixing a cell membrane permeable DNA dye (acridine orange) with diluted whole blood sample that specifically stains white blood cells. To quantitatively analyze WBCs, we simply pipette 20µl of treated sample into a Cellometer Disposable Counting Chamber and place the chamber into the instrument. Since the counting chamber is disposable, no washing is required between samples, there is no risk of cross contamination of different samples, and the risk of exposure to biohazard materials is reduced.

Using proprietary algorithms, Cellometer Vision's robust operating software accurately analyzes cell images, and generates counting data in less than 60 seconds typically. Cell images and all analysis data, including cell size distribution histograms, can be saved for documentation. Data can also be easily exported to Microsoft Excel spreadsheets for further analysis.

Fluorescence cell count and concentration for white blood cells in whole blood
Counting results box displays fluorescence cell count & concentration.


Direct AO labeling of white blood cells:
  1. Collect 1 drop of peripheral blood sample (20-50µl).
  2. Take 20µl of blood sample and dilute the sample with 180µl of PBS.
  3. Apply 5µl of Acridine Orange solution to 100µl diluted
    blood sample (final concentration 2-10µg/ml).
Running Assay:
  1. Load 20µl of sample into the disposable counting chamber
  2. Insert chamber into Cellometer Vision
  3. Select Assay from drop-down menu
  4. Enter Sample ID for this sample
  5. Preview cell images and click 'Count' to begin analyzing sample
  6. Review images and counting data.
  7. Save or export images and/or report data.
Select WBC in Whole Blood Assay

Step 3: Choose Assay from drop down menu

Step 4: Enter Sample ID


AO Stained WBC’s are clearly visible in the acquired fluorescence image (Figure 1). The software indicates counted cells with a green circle (enlarged to show detail). Results display indicates fluorescence cells counted and automatically calculates concentration.

Acridine Orange stained white blood cells
Figure 1. Fluorescent image showing counted AO
stained WBCs. RBCs are not visible.
Cell and fluorescent size distribution histograms, scatter plots and data files can also be instantly generated. All experimental data can be instantly transferred to an Excel spreadsheet (Figure 2) or saved in a data table (Figure 3).

Example of data from AO stained white blood cell assay
Figure 2. Example of data output to Excel.
data table for white blood cell assay
Figure 3. All counting results can be instantly saved to a data table.

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