Dilution of semen

DILUTION OF SEMEN

  • The French mini/medium straw is packed with 20 million sperms.
  • The final dilution is calculated based on volume, concentration and packing volume.
  • Till the final dilution rate is decided, 1:1 diluted semen has to be kept in water bath of 34 ⁰ C.
  • This time period will also help to adjust and stabilize the semen with the diluents.
  • Then it is placed outside the water bath at room temperature (20-22 ⁰ C).
  • Here it is placed for 10 minutes to bring the 1:1 diluted semen temperature to 20 ⁰ C.
  • The final dilution is done at room temperature of 20-22 ⁰ C.

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Example

Calculation of dilution rate

  • For French mini straw
    • The French mini straw is having 20 million sperms in 0.25 ml.
    • Let us consider the

Equation

  • The total diluted volume is 62.5 ml which includes the semen volume also.
  • So to the 5 ml semen 57.5 ml diluents is added to make the final volume of 62.5 ml.
  • The semen is already diluted with 1:1 dilution rate ie to 5 ml of semen 5 ml diluent was already added.
  • So the final volume available is 10 ml. So 52.5 ml of diluent has to be added as per the dilution rate.
  • In a sterile beaker (of 100 ml) add 40 ml of diluent. To this add the 1:1 diluted semen ie 10 ml.
  • The remaining 7.5 ml diluent is added to the collection vial and it is poured into the beaker.
  • When the diluents or semen is poured into the beaker it has to be passed via the side walls of the beaker.
  • It should not be poured directly into the beaker. The air bubble formation should be avoided while dilution is done.

View video (Click here for video demonstration)

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Last modified: Wednesday, 6 June 2012, 2:10 PM