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

WBCs
Cellometer Vision
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Introduction

Brightfield image of whole blood.
Brightfield image of whole blood.
WBC's are not visible amongst RBC's

Fluorescent image showing AO stained WBC's
Fluorescent image showing AO
stained WBC's. RBC's 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 20ml 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.

Counting results box
Counting results box displays fluorescence cell count & concentration.

Method

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.
Steps

Step 3: Choose Assay from drop down menu




Step 4: Enter Sample ID

Results

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.

Fluorescent image
Figure 1. Fluorescent image showing counted AO
stained WBC's. RBC's 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
Figure 2. Example of data output to Excel.
Counting results
Figure 3. All counting results can be instantly saved to a data table.


Schedule an Online Demo
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