1- 200mg epididymis + NaCl till 2ml in a Perti dish (30 mm).

2- Macerate the epididymis by scissor into 4 parts.

3- Leave the tissue for 30-60 seconds to let sperms leak from the tubules (the solution will tint
whitish/grayish, similar to the diluted semen), and the resulted fluid will be handled exactly as
the semen.

4- Collect the fluid only into an Eppendorf tube and perform the semen analysis procedures,
motility, live/dead & abnormalities and sperm cell count.

P.S. for sperm cell count, you first diluted the tissue 10 times, then you will dilute in a
NaCl+formaline (20 microL semen + 180 microL NaCl+formaline) to kill the sperms. so
consider that two dilution factors in final calculation using the hemocytometer chamber. e.g. if
you count 100 sperm in a primary square so the Sp.C.C = 100x10x10x10x1000 = 100
million/ml.

Hope that useful.

If you are currently in Egypt, I sent my phone No. to you as a message.

Islam

Is there a technique for counting the sperm number in rat semen relative to the epididemal
tissue?. Available from:
https://www.researchgate.net/post/Is_there_a_technique_for_counting_the_sperm_number_in_ra
t_semen_relative_to_the_epididemal_tissue [accessed Apr 3, 2017].

Rat sperm count using SMT(Saline Merthiolate Triton X-100). THIS PROCEDURE ONLY TO
BE MADE AT 37ºC.
1. Surgery is made and excise left/right testis and weigh. Identify, isolate and excise
epididymis part and weigh.
2. Isolate and excise caudal part of epididymis and weight.
3. Place cauda epididymis in a container containing 10 ml of SMT.
(SMT is prepare by mixing 0.9%(9g/L) NaCl, 0.01%(100mg/L) Merthiolate, and
0.05%(0.5ml/L) of Triton X-100 in dH20)
4. Homogenize for 1 minutes
5. Take an aliquot of 50µl of homogenate(SMT-homogenized sample) and add 950µl of
10% neutral buffered formalin and vortex thoroughly.