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IBEX Sciences Spermatozoa Count Solution (Semen Diluting Fluid) is a ready-to-use in vitro diagnostic (IVD) reagent designed for manual or semi-automated semen analysis. It functions primarily as a specialized diluting fluid used to accurately determine sperm concentration under a microscope using a hemocytometer (such as an Improved Neubauer counting chamber). [1, 2, 3]
Key Functions & Mechanism
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- Immobilization: The solution contains immobilization agents (typically a mild fixative like formalin) that immediately stop the progressive movement of sperm cells. This ensures they stay stationary within the counting grid, preventing double-counting or tracking issues. [1, 2, 3, 4]
- Optimal Dilution: It thins the thick viscous semen matrix, separating densely packed spermatozoa so individual cells can be easily distinguished and counted under a 40x microscope objective. [1, 2, 4, 5]
- Debris Cleansing: Typical chemical components (such as sodium bicarbonate) assist in clearing mucus and background seminal debris, providing a clean optical field for the technician. [1, 2]
General Procedure for Manual Counting
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- Liquefaction: Allow the raw semen sample to completely liquefy at room temperature or in an incubator (37°C) for 15–30 minutes before mixing. [, 2, 3, 4, 5]
- Dilution: Pipette a precise micro-volume of the well-mixed semen sample and dilute it with the IBEX Spermatozoa Count Solution. A common diagnostic protocol uses a 1:20 dilution ratio (e.g., 0.02 mL semen into 0.38 mL diluting solution). [1, 2]
- Loading: Thoroughly mix the diluted specimen and load a small droplet onto the edge of a clean Neubauer counting chamber, allowing it to draw in via capillary action. [1, 2]
- Microscopy: Let the chamber sit for a couple of minutes to let the cells settle. Count the immobilized spermatozoa across the designated grid areas to calculate millions of sperm per milliliter. [1, 2, 3, 4, 5]
Storage and Handling
- Storage: Keep the reagent bottle tightly capped and store at standard room temperature or according to the label instructions to prevent evaporation.
- Contamination Control: Avoid inserting unsterilized pipettes directly into the primary reagent bottle. Microbial or particulate contamination can be mistaken for cellular artifacts during microscopic valuation. [1, 2]
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