Semen microscopy experiment
After the semen bath is liquefied, take 1 drop of the mixed semen specimen on a clean slide with a coverslip, and then observe the presence of spermatozoa and the activity of spermatozoa under low magnification. If spermatozoa are not seen, centrifuge the specimen and determine whether there are spermatozoa in the sediment. If there are no spermatozoa in the two slices, then no spermatozoa should be reported, and no other tests should be performed. This experiment is from Mudanjiang Medical College, undergraduate 5-year laboratory guide for testing majors.
Operation method
Semen microscopy experiment
Principle
I. Sperm motility test: Observe the wet film with or without spermatozoa with low magnification, then turn to high magnification and observe at least 100 spermatozoa, the percentage of spermatozoa with tail movement is the motility rate, which is calculated by the formula: Sperm motility % = (number of motile spermatozoa / number of motile spermatozoa tens of inactive spermatozoa) x 100 II. Sperm motility test: While checking the motility rate of the spermatozoa, motility is checked according to the following standard Grade 0, inactive, dead sperms Grade I, inactive, sperms are sluggish, rotating in place Grade II, active, good, sperms are active, but not in the same direction Grade III, active, good, sperms are wired, active and moving, normal semen 1 hour after sperm discharge, Grade II, sperms 60%, Spermatozoa morphology 1, semen with a sperm density of >15x109/L; semen with liquefied semen may be directly coated, while semen with a sperm density of <15x109/L may first be liquefied. Semen 1500r/min centrifugation for 10 minutes, take the precipitate smear. 2, the smear will be dried naturally in the air, formaldehyde fixation for 5 minutes, with HE staining, 3, to be stained after drying, in the oil microscope count 200 spermatozoa, report the rate of normal abnormalities. 2 morphology observation of the normal spermatozoa have a head, the middle section with the foot, HE staining the head is purple, the tail is white or purple. Sperm density and total sperm count 1. Rough estimation method: mix the liquefied semen and smear it directly, add a coverslip and observe 5 fields of view under high magnification microscope (40x), and take the average of x109, i.e. the approximate number of sperms per liter. Sperm reading I. Experimental principle: NaHCO3 is used to destroy the viscosity of semen, formaldehyde is used to immobilize spermatozoa, and the semen is quantitatively diluted for counting.
Materials and Instruments
Semen dilution Move Experimental reagents: semen diluent NaHCO3 5g concentrated formaldehyde solution 1ml, distilled water to 100ml. For more product details, please visit Aladdin Scientific website.
NaHCO3 Concentrated formaldehyde solution Distilled water
Experimental operations:
1. Take 0.38ml of diluent in a small test tube.
2. Pipette 20 microliters of the liquefied and mixed semen with hemoglobin Pr and add it to the diluent, and dilute it 20 times.
3. Mix the diluent well and fill it into the cell counting chamber. 4. Leave it for 5 minutes for all the sperm to be deposited, and then count them with a high magnification lens. Fill the cell counting chamber.
4. Leave for 5 minutes to allow all sperm to be deposited, and then count the number of sperm in the corners of the large central square and in the center of the center of the 5 middle squares with high magnification.
Experimental calculations: sperm count/L = number of sperm in the 5 center squares x 109