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Sperm morphology rapid dyeing reagent and method for rapid dyeing sperm

A rapid staining and morphological technology, applied in the field of sperm morphological rapid staining, can solve the problems of difficult routine items, complicated formula composition and long operation time in sperm morphological detection, and achieves easy and wide application, good effect and fast operation. Effect

Inactive Publication Date: 2010-09-15
深圳华康生物医学工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the methods for sperm morphology detection mainly include hematoxylin-eosin staining, modified Papanicolaou staining and Diff-Quik method. The hematoxylin-eosin staining method and the modified Papanicolaou staining method are brightly stained, and each part is easier to distinguish The effect is good, but its composition is complicated, the operation time is long, and it is difficult to be widely used in clinic; the Diff-Quik method is good for sperm staining, and the operation is simple and fast, but its raw materials are expensive and difficult to purchase
[0004] Due to the limitations of existing detection methods, sperm morphology detection is difficult to carry out as a routine project

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1, a kind of rapid staining reagent of sperm morphology, comprises solution A, solution B and solution C, wherein solution A is the mixed solution that contains Nile blue 0.1% acetone, methanol, 40% formaldehyde, is used for staining sperm top Body area; Solution B is the PB buffer that contains Jiaoning Y0.019%, sodium azide 0.1%, phosphate (PB) content is 0.4mol / L, PH is 7.4, is used for staining sperm body and tail area; Solution C is a PB buffer containing 0.1625% basic methylene blue, 0.1625% methylene blue, 0.1% sodium azide, 0.4 mol / L PB, and PH 7.4, and is used for staining the post-acrosomal region of sperm.

[0018] The preparation method of above-mentioned sperm morphology rapid staining reagent comprises the steps:

[0019] A, the manufacturing process of PB buffer solution

[0020] Prepare 1000 ml of 0.2M potassium dihydrogen phosphate and 1000 ml of 0.2M disodium hydrogen phosphate, mix the above two solutions under the monitoring of a pH meter...

Embodiment 2

[0027] Embodiment 2, a kind of sperm morphology rapid staining reagent, comprises solution A, solution B and solution C, wherein solution A is the mixed solution of acetone, methanol, 40% formaldehyde containing Nile blue 0.05 ~ 0.5%, is used for staining Sperm acrosome area; solution B is a PB buffer solution containing 0.01-0.05% coyne Y 0.01-0.05%, sodium azide 0.1-0.2%, PB content 0.5mol / L, pH 7.4, used for staining sperm body and tail area; solution C is a PB buffer solution containing 0.1-0.5% of basic methylene blue, 0.1-0.5% of methylene blue, 0.1-0.2% of sodium azide, a PB content of 0.5mol / L, and a pH of 7.4, and is used for staining the top of sperm posterior area.

[0028] The preparation method of above-mentioned sperm morphology rapid staining reagent comprises the steps:

[0029] A, the manufacturing process of PB buffer solution

[0030] Prepare 1000 ml of 0.15~0.25M potassium dihydrogen phosphate and 1000 ml of 0.15~0.25M disodium hydrogen phosphate, mix the...

experiment example

[0039] In the experimental example, the specific steps for rapid staining of sperm are as follows:

[0040] 1. Sample preparation

[0041] Drop a small drop (5~10ul) of fresh liquefied semen on a clean slide, and use the edge of the second slide to push forward at an included angle on the surface of the slide to make a smear of appropriate thickness.

[0042] 2. Dyeing steps

[0043] 1. After the sperm suspension smear is prepared, place it horizontally and let it dry naturally in the air;

[0044] 2. Add solution A to cover the smear and react at room temperature for 15 seconds, and stand the slide on absorbent paper to remove excess dye, which is used for staining the acrosome area of ​​the sperm suspension smear;

[0045] 3. Add solution B to cover the smear and react at room temperature for 10 seconds. Stand the slide on absorbent paper to remove excess dye, and use it to stain the body and tail of the sperm suspension smear;

[0046] 4. Add solution C to cover the smea...

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PUM

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Abstract

A sperm morphological fast stain consists of solution A containing Nile blue fixation liquid of tissues for staining sperm apical body region. A method for fast staining includes preparing sperm suspension film preparation, drying by air, using said solution A to cover film preparation till sperm apical body region be stained.

Description

technical field [0001] The invention relates to an in vitro diagnostic reagent, which is particularly suitable for rapid staining of sperm morphology. Background technique [0002] Sperm morphology detection plays an important role in the evaluation of male fertility and the selection of assisted reproductive methods. [0003] At present, the methods for sperm morphology detection mainly include hematoxylin-eosin staining, modified Papanicolaou staining and Diff-Quik method. The hematoxylin-eosin staining method and the modified Papanicolaou staining method are brightly stained, and each part is easier to distinguish The effect is good, but its formula is complex, the operation time is long, and it is difficult to be widely used in the clinic; the Diff-Quik method stains sperm well, and the operation is simple and fast, but its raw materials are expensive and difficult to purchase. [0004] Due to the limitations of existing detection methods, sperm morphology detection is ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/31G01N1/30
Inventor 傅剑华刘瑜胡家纯何林何小红
Owner 深圳华康生物医学工程有限公司
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