Spermcounting

Estimation of sperm concentration

with the improved Neubauer haemocytometer

The central grid of the improved Neubauer haemocytometer contains 25 squares in which the spermatozoa are to be counted

Dilutions and conversion factors for the improved Neubauer haemocytometer

conversion factors

Number of
large squares
counted

estimated spermatozoa per field ( GX400) Dilution (semen + diluent) 25 10 5

<15 1:5 (1+4) 20 8 4

15-40 1:10 (1+9) 10 4 2

40-200 1:20 ( 1+19)
5
2 1

>200 1:50 (1+49) 2 0,8 0,4

Preliminary precautions before counting:

.Let semen sample liquifying (15 to 30mn)

homogenize by aspirating up & down through a 5 ml syringe

perform dilution

add 10µl of diluted samples into both chambers (coverslide should perfectly stick to the haemocytometer, observe the Newton rings!)

count only spermatozoa with a tail (double counting ,2 chambers)

mentionany agglutinates , mucous threads ,aggregations of spermatozoa, abnormal viscosity in the analysis report

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			conversion factors
		Number of	large squares	counted
estimated spermatozoa per field ( GX400)	Dilution (semen + diluent)	25	10	5
<15	1:5 (1+4)	20	8	4
15-40	1:10 (1+9)	10	4	2
40-200	1:20 ( 1+19)	5	2	1
>200	1:50 (1+49)	2	0,8	0,4