Sperm morphology staining kit

【Sperm morphology staining kit(Pasteur Rapid Method)Product Name

Sperm five test:Sperm morphology staining kit(Pasteur Rapid Method)

【Research and Development Basis

The lower the proportion of normal shaped sperm in semen, the lower the chance of reproduction, and the greater the possibility of miscarriage, fetal dysplasia, and stillbirth; Data from assisted reproductive technology shows that the sperm deformity index (SDI) is associated with in vitro fertilization, and when SDI>1.6, the rate of in vitro fertilization decreases; The multiple abnormalities index (MAI) and teratozoospermia index (TZI) are associated with in vivo fertilization. The World Health Organization (WHO) points out that when the normal sperm count is ≤ 4%, it will have a significant impact on male fertility.

Mainly used for staining human sperm morphology, it facilitates doctors to observe the normal morphology of sperm under a microscope and calculate the proportion of defective sperm, evaluating the quality of human sperm morphology. It can also be used for morphological examination of sperm in other mammals.

No need for special instruments, simple, fast, and accurate.

【Principle of Inspection

This staining method is the Babbitt staining method recommended by WHO. Due to the fact that sperm and proteins with different isoelectric points in cells carry different charges at the same acidity, they can selectively bind to corresponding dyes and exhibit different colors. The nucleus is composed of acidic substances and has a strong affinity for alkaline dyes; On the contrary, cytoplasm has a higher affinity for alkaline substances and acidic dyes. This kit can be used to evaluate the morphological characteristics of human sperm and can also be used for morphological examination of sperm in other mammals.

【 Composition 】The raw materials for reagent preparation include hematoxylin, eosin, phosphotungstic acid, bright green, glacial acetic acid, and alcohol.

Fresh semen sample after liquefaction

【Operation method

1. Sample preparation

（1）Fresh semen samples are liquefied or stored at -20 ° C for thawing (room temperature).

（2）Sperm needs to be washed and sliced as follows: ① Add semen into a test tube0.5mlAnd physiological saline solution10 mlMix thoroughly. ②2000turncentrifugation10minRemove most of the supernatant (repeat steps ①② if necessary). ③ Gently flick the test tube to suspend the centrifuged sperm mass in the remaining saline solution. If the sperm concentration is too high, adjust the sperm density appropriately with physiological saline solution.

2Testing steps

（1）On a clean glass slide, add 1020μlSemen, evenly spread it into a circular sample area, place the coated glass slide horizontally, and let it dry naturally in the air.

（2）Fix the dried smear in a 95% ethanol solution for 510 minutes.

(3) Soak the fixed smear in dye solution A for 5 minutes and wash with water.

(4) Immerse the smear in dye solution B for 5-10 seconds, wash twice with water, and then soak in clean water for 2 minutes.

(5) Immerse the smear in 95% ethanol for 2 minutes.

(6) Soak the smear in dye solution C for 3 minutes and rinse twice in 95% ethanol (510 seconds each). After drying, add paraffin oil dropwise (if the specimen needs to be stored for a long time, it needs to be sealed with "neutral gum"), and observe under an optical microscope at least200Individual sperm, classify and count the morphology of sperm.

【 Staining Results 】

The acrosome area of sperm is light blue, the non acrosome area of the head is dark blue, and the tail of the sperm is red.

（1）WHOClassification criteria for sperm morphology: Normal sperm: elliptical head (normal size, aspect ratio)1.501.75The boundary of the top body is clear, and the top body occupies the head4070%）The head, neck, midsection, and tail are all normal.

(2) The classification of defective sperm is as follows:

Calculation of multiple sperm defect indicators: ① Multiple abnormality index（the multiple anomalies index, MAI）The average number of defects in each abnormal sperm, including all head, neck, and tail deformities. ② Abnormal sperm index（teratozoospermia index，TZI）：TZIandMAISimilar, the difference between the two lies inTZIOnly record the four types of defects in each sperm: head, neck, tail, and whether there are excessive residual cytoplasmic droplets, rather than the actual number of defects in each abnormal sperm. ③ Sperm abnormality index(sperm deformity index，SDI)The number of defects divided by the total number of sperm (not just the number of abnormal sperm). This index can record various abnormalities in the head of sperm, but defects in the neck and tail are only recorded once.

The calculation method for multiple sperm defect indicators is as follows:

(3) World Health Organization（WHO）Recommended normal reference value:Normal form sperm﹥4%

【Precautions:

1、Insufficient sperm washing will affect microscopic observation.

2、Participants should follow theWHOTechnical specifications for collecting semen samples.

3When dyeing in winter, if the ambient temperature is low, the dyeing time of dye A and dye C can be appropriately extended.

4All samples of the test subject should be considered as potential sources of infection, and the tested samples, reagents, and utensils must be treated as medical waste to avoid environmental pollution.

5The sperm staining kit is an in vitro diagnostic reagent, and the test results are for clinical reference only. The clinical management of patients should be considered in conjunction with their symptoms/signs, medical history, other laboratory tests, and treatment response.

【 Basic Information 】

Product registration certificate number: Minming Food and Drug Administration (Approval) No. 2013 1400021

Execute product standard: YZB/Minming 0033-2013

Medical Device Manufacturing Enterprise License No.: Min Shi Yao Guan Xie Production License No. 20110297

Packaging specifications: 30ml x 6 bottles/box, 100ml x 3 bottles/box, 250ml x 3 bottles/box, 500ml x 3 bottles/box

Storage conditions and expiration date: The reagent kit is available from 4-28℃light-proofSave, valid for 12 months.

[Appendix]

Factors affecting sperm fertilization or male infertility

1.High sperm mortality rate

2.High sperm abnormality rate

3.Low sperm maturation rate

4.Chromosomal abnormalities in sperm

5.Insufficient sperm count

6.Low sperm motility

7.Low sperm liquefaction ability

Sanming Zhonghe Biotechnology Co., Ltd. Reproductive Medicine Testing Kit Series

1Sperm DNA detection kit

2Sperm DNA Fragment Detection Kit

3Sperm Activity Test Kit

4Sperm morphology detection kit

5Sperm Nuclear Protein Detection Kit

Five joint tests are used to diagnose and analyze the fertilization and fertility of sperm, without the need for instruments. They are simple, fast, and accurate,Infertility DepartmentThe diagnostic plan.