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Cat.No: FS-1622 Datasheet Instruction for Use
Specification Quantities
Price $511.77–852.95
| Product Name | Sperm Staining Kit (Aniline Blue-Eosin Staining) |
| Catalog No. | FS-1622 |
| Description | Under normal conditions, the basic proteins (nucleoproteins) that bind to sperm nuclear DNA undergo a natural maturation process from histones to spermin. This matured spermin provides special protection for sperm DNA. The process by which histones are gradually replaced by spermin is called sperm nucleoprotein profile conversion, which has significant physiological importance. Therefore, the amount of histones is an important indicator of sperm maturity. The principle behind the sperm nucleoprotein staining solution (eosin-aniline blue method) is that, under acidic conditions, aniline blue specifically binds to lysine residues abundant in sperm nuclear histones to form a violet-blue compound. The degree of sperm maturity is determined based on the intensity of the staining. |
| Test Procedure | Preparation of the washing buffer: Take an appropriate amount of 10× concentrated washing buffer and dilute it 10-fold with distilled water to prepare the washing buffer. Preparation of the elution buffer: Take an appropriate amount of 10× concentrated elution buffer and dilute it 10-fold with distilled water to prepare the elution buffer. Place 0.2–0.5 mL of liquefied semen into an EP tube, then add 1–1.5 mL of washing solution. Pipette up and down several times using a pipette or syringe, centrifuge at 2000 rpm at room temperature for 5 minutes, discard the supernatant, and retain the sperm pellet at the bottom of the tube; repeat this procedure three times. Add approximately 0.1–0.2 mL of the working wash solution to the EP tube from Step 3 to prepare a mixed sperm suspension. Take 15 μL of the prepared sperm suspension and spread it evenly on an anti-smear slide; allow to air dry. Add 2–3 drops of fixative to the area of the slide containing the sperm to be tested, fix at room temperature for 5–15 minutes, rinse with distilled water for 5–10 minutes, and remove excess water by centrifugation. Add 2–4 drops of aniline blue staining solution to the smear area, stain at room temperature for 5 minutes, rinse with distilled water for 5 minutes, and shake off excess water. Insert the slide vertically into the reaction well containing the elution buffer, ensuring the smear area of the specimen is completely submerged in the elution buffer, and incubate at room temperature for 2–5 minutes. Rinse with distilled water for 1–5 minutes, then remove excess water by centrifuging. Add 2–4 drops of counterstain solution to the smear area, stain at room temperature for 2–5 minutes, and rinse with distilled water for 1–5 minutes. Quickly air-dry the slide and examine under a microscope. If long-term storage of the sample is required, sequentially place the slide in 70%, 80%, 95%, and 100% ethanol for 2 minutes each, then clear with xylene and mount with neutral resin. |
| Storage and Transportation | Store at room temperature, protected from light. Shelf life: 6 months. |
| Precautions | When rinsing the slide with distilled water, be sure to control the flow rate to prevent the sperm in the smear from being washed away. When removing excess water, take care to prevent the smear from drying out. For your safety and health, please wear a lab coat and disposable gloves when performing this procedure. |
For research use only, not for clinical use.
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