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question and inquiry

By: talha ,
02 September 2009
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Question:

Hi ,

  my name is Talha , currently working as Embryologist in a pharma , I want to ask that how will be sperms counted if there are 40 -50  million sperms phpf i mean will the sperms  be in thousands expressed, i.e what wil be count actually if the count is mentioned bove ,  i am actually not been able to diffrentite 40-50 million per ml and phpf how wil I count it .

The other one is regarding Halo Sperm kit , can some body give me technical information , about it i am going to start to work on the kit and i have no prior experience on it ,thank you

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Michael L Reed



Michael L Reed said on 02 September 2009

"A few comments on sperm counting using the sperm per hpf method. From the ESHRE Monographs Manual on Basic Semen Analysis, June 2002, in the section discussing sperm concentration, the text states that 4 sperm per high powered field corresponds roughly to a concentration of 1 m/ml. So if you are counting 40 to 50 sperm phf, then you could assume that the concentration would be approximately 10 to 13 m/ml. This is also assuming that you are using 400X magnification, given that your field of vision is 500 micrometers in diameter, and given that you are using 6 microliter volume of well mixed semen under an 18x18 coverslip on a glass slide. The text states also that older microscopes may have different diameter fields of vision. So, there is a lot of room for error using this method. Yes, this method is likely ok for qualitative semen analysis - a screening method perhaps. But I would consider an alternate method for more accuracy and precision.

The second part to your question regarding the Halosperm kit, this test allows the user to perform sperm decondensation testing, reflecting sperm DNA fragmentation, in-house, using either brightfield microscopy or flurorescent microscopy. The test is fairly easy to perform, with very good step-by-step instructions. Reading the slides is like reading a version of a stained morphology, so you can expect to have a learning curve, with refinement of inter- and intra-technician variation over time. As more programs use the test, and compare the results to clinical outcomes, and assuming we can share information between all of us, then the true utility of this assay can be determined. Like any clinical test, you'll have to decide how best to use the data in your setting.

Mike"

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