HEMOCYTOMETRY

BANAG, Julie Ann S.

GAMBOA, Lois Danielle L.
PAULE, Diosshane V.
BSMT 3 C
Group 9

The device used for determining the number of

cells per unit volume of a suspension is called a
counting chamber. It is the most widely used type
of chamber, since it was mainly designed for
performing blood cell counts. It is now used to count
other types of cells and other microscopic particles as
well.

The hemocytometer was invented by Louis-Charles

Malassez. It is a special type of microscope slide
consisting of two chambers, which is divided into
nine (1.0mm x 1.0mm) large squares which
are separated from one another by triple lines. The
area of each is 1mm. Cover glass is supported over the
chambers at a height of 0.1mm. Because of that the
entire counting grid lies under the volume of 0.9 mm
on one side. The cell suspensions are introduced into the
cover glass. The hemocytometer is placed on the
microscope stage and the cell suspension is counted.

 counting chamber is not clean

cover glass is not placed correctly onto the
chamber
chamber is not filled without bubbles
chamber is overfilled
not enough time for sedimentation of the cells
the wrong cover slip is used. Standard cover
slips for microscopy are too thin and bend
because of the capillary forces.

RESEARCH QUESTIONS

a.

b.

Number of squares in one ruled area:

Area and volume of the one ruled area: 0.25 x 0.25 mm

(0.0625 mm^2) for area, 0.00625mm^3 for volume

c.

Area and volume of one large square: 1.0 x 1.0 mm

(1mm^2) for area, 0.1mm^3 for volume

d.

Depth or volume of one counting chamber: 0.1mm

a.

Number of squares used in counting:

b.

Total area and volume used in counting:

4mm^2 for area, 0.4mm^3 for
volume

corner squares

a. Number

b. Total

of squares used in counting:

area and volume used in counting:

2mm^2 for area, 0.2mm^3 for
volume


Color of
Bead(s)
Dilution Range
Markings

RBC Diluting
Pipette

WBC Diluting
Pipette

Red

White

1:100 to 1:1000
0.5, 1.0 and
101.0

1:10 to 1:100
0.5, 1.0 and 11.0

REFERENC
ES:
http://ruby.fgcu.edu/courses/hogedegb/80026/hem01lab1a.htm
http://www.free-ed.net/free-ed/Courses/06%20MedHealth/MD
%20Series/MD0853/Hematology.asp?iNum=0202
http://www.free-ed.net/sweethaven/MedTech/Hematology/lessonMain.asp?iNum=0204
http://vlab.amrita.edu/?sub=3&brch=188&sim=336&cnt=1
http://www.microbehunter.com/the-hemocytometer-counting-chamber/
http://www.hausserscientific.com/products/speirs_levy_eosinophil.html