Solarbio Sperm Morphology Stain Kit(Diff-Quik)

Kit Components

Before use, mix Reagent (A):(B):(C) in a ratio of 1:1:1 to form Diff-Quick Staining Solution. Use it immediately; do not store for a long time.

Uvod

Diff-Quick Staining is a rapid staining method based on improvements to the Wright Staining technique. It is commonly used in cytology tests. The dye solution is prepared using a rapid staining method recommended by the WHO. Similar to Wright Stain, it is based on the principles of Romanowsky Stain technology. The staining results are very similar to those of Wright Stain, but Diff-Quick Stain takes a very short time, usually within 90 seconds. Diff-Quick Stain can be used to evaluate sperm morphology.

Sperm Morphology Stain (Diff-Quick Method) is a sperm morphology staining solution recommended by the WHO. Its dyeing principle is based on the fact that sperm and proteins with different isoelectric points in cells have different charges under the same acidity. They can selectively combine with corresponding dyes for staining. The dye is mainly used for sperm (cell) morphology evaluation and is only intended for scientific research, not clinical diagnosis or other purposes.

Self Provided Materials

Slide, Distilled Water, Microscope.

Protocol (For reference only)

1. Prepare a smear by washing two slides thoroughly, then washing with 70% ethanol and drying. Add 5-20μL onto the slide, drag a drop of semen on the surface of the clean slide with the edge of the second slide, and make a smear. If the sperm density is too high, it can be diluted with physiological saline.
2. Put the smear in Diff-Quick Fixative or let it dry naturally, and fix for 15-20 seconds.
3. Place the smear vertically on absorbent paper to remove excess liquid.
4. Stain with Diff-Quick I for 10-20 seconds and place the smear vertically on absorbent paper to remove excess liquid.
5. Stain with Diff-Quick II for 5-10 seconds and place the smear vertically on absorbent paper to remove excess liquid.
6. Wash with running water 10-15 times to remove excess dye solution.
7. Place the smear vertically on absorbent paper to remove excess liquid and allow it to completely dry. View under the microscope.

Result

• Acrosome region of sperm head: Light Blue
• Postacrosomal region: Deep Blue
• Middle part: Maybe Slight Red
• Rear part: Blue or Slight Red

Note

1. Adjust the volume of sperm sample based on density.
2. Diff-Quick Fixative is recommended, but methanol can be used as a substitute.
3. Use fresh sperm samples and ensure an even and thick smear for optimal staining.
4. Avoid removing dye solution directly or washing the smear vigorously to prevent settling.
5. Reuse dye solution cautiously and filter if sediment occurs.
6. Decolorize with methanol if necessary, avoid re-staining.
7. Adjust staining time or concentration if colors are too deep or shallow.
8. Ensure clean slides to avoid affecting staining due to pH alterations.
9. Wear appropriate safety attire and disposable gloves.