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WBC Count: Hemacytometer
WBC Count: Hemacytometer
HEMACYTOMETER
The hemocytometer is a device originally designed and usually used for counting blood cells.
The hemocytometer was invented by Louis-Charles Malassez and consists of a thick glass
microscope slide with a rectangular indentation that creates a chamber.
Materials
The necessary elements to perform a cell count with Neubauer chamber are as followS:
A. cellular dilution to measure
B. hemocytometer, or Neubabuer chamber
C. optical microscope
D. cover glass
E. pippette / micropippete with disposable tips.
F. dilutlon buffer / PBS (if needed)
THE NEUBABUER CHAMBER, OR HEMOCYTOMETER
Double chambers are most common than simple chamber. In this case, the chamber has two counting
areas than can be loaded independently.
Neubauer chamber's counting grid is 3 mmx3 mm in size. The grid has 9 square subdivisions of width
1mm.
Counting Grid
IMPROVED NEUBAUER CHAMBER COUNTING AREA
Counting Grid Areas Are present on central platform of chamber
Counting Grid
Each scale is 3mm wide and 3mm
long (9mm2 total area)
The whole scale is divided into 9
big squares.
Each squaree 1mm long and 1mm
wide. (1mm area)
In case of blood cell counting, the squares placed at the corners are used for white cell counting. Since
their concentration is lower than red blood cells a larger area is required to perform the cell count
The central square is used for platelets and red cells. This square is split in 25 squares of width
0.2 mm (200um).
Each one of the 25 central squares is subdivided in 16 small squares
WBC count = cells counted X dilution factor
Area counted (mm2) X depth
= 96 X 20
4 X 0.1
= 4800/mm3 or
4 800/µL or
4.8 X 10^3/µL or
4.8 X 10^9/L