The Makler Counting Chamber

Chamber Description: The Makler Counting Chamber is only 10 microns deep: 1/10th of the depth of ordinary hemocytometers, making it the shallowest of known chambers.  Constructed from two pieces of optically flat glass, the upper layer serves as a cover glass, with a 1 sq.mm fine grid in the center subdivided into 100 squares of 0.1 x 0.1 mm each.  Spacing is firmly secured by four quartz pins. 

Analysis Technique: A small, uncalibrated drop from a well mixed undiluted specimen is placed in the center of the Chamber by means of a simple rod and immediately covered.  A microscopic objective of x20 is required. 

Motility Evaluation: Non-motile sperm are counted within an area of nine or sixteen squares in the center of the grid.  Moving sperms are then counted, and graded if desired.  The procedure is repeated in several areas.  Percentage of motility and its quality are then calculated. 

Sperm Count: A part of the original specimen is transferred to another test tube for immobilization by placing the tube in hot water (50o-60o): a cup with 2/3 boiling water and 1/3 tap water is suggested.  A drop of the immobilized specimen is then placed in the Chamber and counting initiated: sperm heads within a ten square area are counted in the same manner as blood cells are counted in a hemocytomer, their number represents their concentration in millions per ml.  In cases of oligospermic semen, sperms in the entire grid area are to be counted, representing their concentration in hundreds of thousands. 

The Chamber is easily rinsed with water for reuse.  Contact surfaces are wiped with special lens paper after washing. 

Advantages: 

• Applied spermatozoa are uniformly distributed and monolayered, and are observed in one focal plane.
• Dilution is unnecessary even with concentrated specimens.  Analysis is done directly from original specimen in its natural environment.
• All spermatozoa acquire friction free,  horizontal movement and are always examined under constant conditions.
• The specimen can be analyzed quickly as an office procedure while the patient is waiting, and even by an inexperienced person.
• Accuracy of analysis is enhanced through the elimination of the various steps required by the usual hemocytometric technique.  In addition, the fact that sperm motility is examined each time under identical conditions further increases accuracy.  Errors incurred by uncontrolled pressure applied to the cover slip are thus avoided.
• The 10 micron depth of the Makler Chamber is ideal for still or movie camera photomicrography, as it approximately matches the field depth of the objective used in semen analysis.
• The Chamber is quickly and easily available for reuse.  In a busy laboratory a large number of tests per hour can be made by a single technician with minimal technical and material requirements.

References:  
Makler, A. : Fertil. Steril., 33:160, 1980.

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