Initial Microscopic Evaluation

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An initial microscopic evaluation is something that is done in the evaluation of semen quality in the diagnosis of male infertility or other reproductive disorders. The initial microscopic evaluation of semen is completed with 100x magnification and it is meant to asses the following statistics: mucus strand formation, sperm aggregation or agglutination. This evaluation also deals with observing cellular elements besides spermatozoa; these cellular elements are represented by round cells, which can be leukocytes and immature germ cells, as well as isolated sperm head or tails.


As we mentioned above, one of the steps of the initial microscopic evaluation entails assessing sperm agglutination; the WHO standard for a precise grading of agglutination is as follows:

  • Grade 1; this represents isolated agglutination, with 10 or less spermatozoa per agglutinate and most of the spermatozoa being free;
  • Grade 2; this represents moderate agglutination, with 10 to 50 spermatozoa per agglutinate and some free spermatozoa;
  • Grade 3; large agglutination presenting more than 50 spermatozoa per agglutinate and with very few free spermatozoa;
  • Grade 4; gross agglutination, the most severe of agglutination forms, with all the spermatozoa agglutinated and all agglutinates interconnected.

The next step in initial microscopic evaluation of the male semen is assessing the motility and estimating sperm concentration; for this measurement to be accurate, the microscope is switched to 400x magnification. Generally, the estimate of sperm concentration is needed to establish the adequate dilution – the dilution that needs to be obtained – for performance or efficiency of the sperm count. The type of microscope recommended for initial microscopic evaluation of the semen, sperm count and motility evaluation is a phase contrast microscope.

Now we will present the correct steps that are followed for a correct assessment of sperm quality. The physician or other employee of the clinic who executes the test needs to thoroughly mix the semen sample, in order to make sure that the part of the specimen that will be examined is representative and illustrative of the whole specimen, or rather of the general aspects of the male’s sperm at all times.

Some physicians recommend the following mixing technique: the tester is supposed to gently aspirate the semen specimen into a disposable wide-bore plastic pipette, making sure no air bubbles are allowed into the mix. This task should be repeated ten times, carefully each time, so that no mistakes are made. Immediately after mixing, the lab assistant needs to purchase the sample needed for the testing and subject it to all the required measurements.