Pre-Stained Morphology Slides For Sperm Morphology Analysis

ASCEN Pre Stained Morphology Slide Intended use:

to assess the morphological parmeters of cells such as spermatozoa. The stained cells can be assessed with them help of an automated system(i.e. CASA) or manually。

Principle of the device:

when a small volume of cells / semen is deposited on a pre-stained slide,the fixative-stain film dissolves in fluid/seminal plasma and stains (sperm) cells。

Preparations before use:

Remove ASCEN pre-stained morphology slide from packaging and take care not to touch the coated area. Make sure the coated area is undamaged. Access toan incubator at 55°C / 131F, or an ethanol flame. Use semen after liquefaction or (sperm) sample in a culture medium。

Fresh liquefied semen or washed semen sample.

1.Pipette 1.5-2uI specimen on the central area of the pre-stained slide;

2.Stir 1-2 circles the semen with pipette tip to gently mix the semen on the slide;

3. Cover with a cover glass (22x22mm or 24x40 mm );

4. Press gently on the cover glass or turn over the slide on the soft tissue and press the back of slide gently to make sure the semen expanse to maximum range;

5. Observation with microscope.

Protocol B:Making smear:

Suitable for SCA CASA system and manual analysis

Fresh liquefied semen or washed semen sample.

1. Pipette 3.5-5uI specimen on the edge of the color area of the pre-stained slide

2. Make the smear by using a blank slide on it and dragging it slowly toward the other end of the glass at an angle of about 45 degrees35-45 thickness semen layer on it

3. Dry by air

4. Drop it into distilled water for 1 second and air dry the side.

5. Repeat step 4.

6. Air dry

7. Analysis by SCA CASA system or manual observation.

Protocol for preparing washed semen and adjusting the concentration of semen with in 45- 60M/mI(Recommended for SCA CASA system analysis )

1.Mix gently of 1mI (relate with the density of sperm) liquefaction semen with 10mI physiological saline or 10mI PBS (pH7.4t 0.2)

2. Centrifugal 5-8min 800g

3. Remove the most of the Suspension

4. Mix the rest completely , (usually a 20~40uI. )

5. Count the concentration of sperm by by SCACASAor Markler Chamber* Conc. = 45M/mI 50M/ ml,perfect go to ProtocolBorA

Conc. =<45M/ml-60M/m,Perfect go to Protocol B or A

Conc. < 45M/m, add 20ul physiological saline or 10mI PBS (pH74 t o.2) repeat step 2.

Conc. > 65M/ml,dilute with physological saline or 10mI PBS (pH7.4? 0.2)