Prepare sample by diluting 1.0 mL sample and 9.0 mL distilled water.

Add 5 drops Methylene blue.

Use 0.1 mL of diluted sample for counting under the microscope. Observe
under scanner, LP & HP. Read/ count under HP.

1 2

X X

5
X

X X

4 3
Haemacytometer 1/10 depth

TBC - Total Budding Count

TDC - Total Dead Count
TCC - Total Cell Count

Top Chamber
5

Bottom Chamber
5
 Mean Count/field = Sum Count of 10 chamber
10 chamber

Ex. TBC = (ave.bud)(2.5x10⁵)(dil’n factor)

0.1 mL

= (9)(2.5x10⁵)(10)
0.1 mL

= 19x10⁶
0.1

= 19x10⁷ or 1.9x10⁸

Dilution factor, df = 1 = 1 = 10
1/10 0.10

1 = 2.5 x 10⁵
2.5 x 10⁻⁵ standard volume of sample in
one square

Actual Formula:

Mean Count = count

field(chamber) ( 1/ 2.5 x 10⁻⁵ )( 1/1/10 )( 0.1 mL )
where:
2.5 x 10⁻⁵ = standard volume of sample in
one square of haemacytometer with
1/10 depth

1 = volume of sample diluted

1/10 ( 1 mL sample 9 mL distilled

Water)
 TCC = ( 65 )( 2.5 x 10⁻⁵ )( 10 )( 10 )
= 130 x 10⁵ ( 10 )( 10 )
= 13,000 x 10⁵
= 13 x 10⁸

If the sample is very concentrated, do the serial dilution:

1
10
1 mL 9 mL H₂O ( 1/10 dilution)

( 0.1 )( 0.1 ) = 0.01

df = 1 = 1
dilution 0.01

= 100