Cell counting using Hemocytometer

Ariola, Rhoanna L.Diamance, Mary Hazel B.

What is Hemocytometer?

Hemo: blood Cyto: cell Meter:

measurement/counter

*Thus, it is an instrument used to count the blood cells.

What is Hemocytometer?

It includes:a) Neubauer’s

slideb) Cover slipc)

micropipetted) RBC pipettee) WBC pipette

Neubauer’s slide

It is a thick glass slide, center of which has double ruling area separated by troughs (these four troughs are extending across the slide and set parallel to each other. The fifth one is separating the two ruling areas from each other).

Cell Counting with Neubauer ChamberStep 1. Put the glass cover on the Neubauer slide central

area. Take 10 µl sample from a suspension using a

micropipette. Introduce it to the Neubauer chamber. When the

pipette is loaded, it must always be held in vertical position.

Place pipette tip close to the glass cover edge, right at the centre of the Neubauer chamber.

Release the plunger slowly watching how the liquid enters the chamber uniformly, being absorbed by capillarity.

In case of the appearance of bubbles, or that the glass cover has moved, repeat the operation.

Cell Counting with Neubauer Chamber

Cell Counting with Neubauer ChamberStep 2. Place the Neubauer chamber on

the microscope stage. Turn on the microscope light.. Focus the microscope until a sharp

image of the cells is already seen. Look for the first counting grid

square where the cell count will start.

1 mm

0.04 mm

Counting Rule

Pattern in counting cells

For bigger squares:(A+B+C+D)(2500)

Cell density is …..

For smaller square:(a+b+c+d+e)(50,000)

a

e

c

b

d