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Bone marrow specimen processing method, decalcification solution, and applications

A technology of specimen processing and processing method, which is applied in the field of medical and chemical engineering, can solve problems such as unclear structure, unclear staining, and unclear slices of bone marrow specimens, and achieve the effect of ensuring integrity and easy tissue sectioning

Inactive Publication Date: 2019-03-05
SHANGHAI SIMPLEGENE CLINICAL LAB CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In view of the above-mentioned shortcomings of the prior art, the purpose of the present invention is to provide a bone marrow sample processing method, decalcification solution and its use, which are used to solve the problem of unclear slices, unclear structures, and unclear staining of bone marrow samples in the prior art. The problem that the decalcification time is difficult to control

Method used

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  • Bone marrow specimen processing method, decalcification solution, and applications
  • Bone marrow specimen processing method, decalcification solution, and applications
  • Bone marrow specimen processing method, decalcification solution, and applications

Examples

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Effect test

Embodiment 1

[0046] Embodiment 1 bone marrow pathological specimen making

[0047] (1) Bone marrow biopsy tissue was obtained (Shanghai Xinpeijing Medical Laboratory).

[0048] (2) Tissue fixation, the first decalcification treatment: add the above-mentioned bone marrow tissue into the decalcification solution and soak for 120min, the components of the bone marrow decalcification solution: 200mL formic acid (99%), 50mL formaldehyde, 52mL glycerol, 200mL distilled water .

[0049] (3) Dehydration (fixed - dehydrated - transparent - soaked in wax) for 12 hours.

[0050] (4) Embedding: the bone marrow tissue was embedded in paraffin.

[0051] (5) Rough repair: to expose the cut surface of the tissue.

[0052] (6) The second decalcification treatment: the bone marrow tissue is added into the decalcification solution for the second decalcification, and the time is 30 minutes. Triol, 400mL distilled water.

[0053] (7) Slicing: the thickness of the slice is about 3 microns, and the slice of...

Embodiment 2

[0058] Example 2 HE staining of bone marrow tissue

[0059] Experimental example and comparative example prepared in embodiment 1 carry out the following operations:

[0060] Staining: ① Paraffin sections were dewaxed in xylene for 3 times, 10 minutes each time. ②Put in 100%, 95%, 85%, and 70% gradient ethanol for hydration, 2 times for each concentration, 2 minutes each time. ③ Stain with hematoxylin for 5 minutes (Besso), rinse with running water for 3-5 minutes, differentiate for 10 seconds, invert blue for 3 minutes, stain with eosin for 5-10 seconds (Besso), dehydrate, transparent, and mount.

[0061] After HE staining, the results were observed with a microscope (BX51). The results of the experiment example are as follows: figure 2 Shown; In the comparative example observation results such as figure 1 shown.

[0062] pass figure 1 and figure 2 visible, figure 2 compare figure 1 The tissue section is complete, with clear cell structure, uniform thickness, no fo...

Embodiment 3

[0063] Example 3 Bone marrow pathological specimen preparation and HE staining

[0064] (1) Bone marrow biopsy tissue was obtained (Shanghai Xinpeijing Medical Laboratory).

[0065] (2) Tissue fixation, the first decalcification treatment: add the bone marrow tissue above into the decalcification solution and soak for 100 min. The components of the bone marrow decalcification solution are: 198mL formic acid, 52mL formaldehyde, 48mL glycerin, and 210mL distilled water.

[0066] (3) Dehydration (fixed - dehydrated - transparent - soaked in wax) for 12 hours.

[0067] (4) Embedding: the bone marrow tissue was embedded in paraffin.

[0068] (5) Rough trimming: use a microtome to expose the cut surface of the tissue.

[0069] (6) The second decalcification treatment: add the bone marrow tissue into the decalcification solution for the second decalcification, the time is 25min, wherein the components of the decalcification solution: 50mL concentrated hydrochloric acid (35%), 25mL ...

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Abstract

The invention provides a bone marrow specimen processing method, a decalcification solution, and applications. The bone marrow specimen processing method comprises following processing steps of bone marrow tissues: fixing and primary decalcification, dehydration, paraffin embedding, rough trimming, secondary decalcification, and slicing; the decalcification solution at least comprises formic acid,formaldehyde, and distilled water at a volume ratio of 198-202:48-52:190-210, wherein the mass concentration of formic acid ranges from 97 to 99%. The bone marrow specimen processing method is used for processing bone marrow specimen, so that slices with relatively complete tissue structures and clear cell structures can be obtained, and it is beneficial for observation of slices under microscopes.

Description

technical field [0001] The invention relates to a bone marrow sample processing method, a decalcification solution and uses thereof, belonging to the field of medical and chemical engineering. Background technique [0002] Bone marrow biopsy (BMB) technology is more and more superior in the diagnosis of bone marrow slices, but bone marrow tissue contains a large amount of mineralized substances, which makes slices very difficult, and bone marrow is the birthplace of various cells, cell types and shapes It is extremely abundant and difficult to diagnose only by routine film production, so it must be combined with immunohistochemical staining to facilitate accurate diagnosis. [0003] Bone marrow tissue is rich in calcium salts and adipose tissue, which must be decalcified first before tissue sectioning. The traditional processing of bone marrow specimens includes the following steps: sampling, fixation and decalcification, dehydration, paraffin embedding, sectioning, HE stai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
CPCG01N1/30G01N2001/305
Inventor 杨文娟周冬冬潘红超赵琨程乐华陈忠黄士昂
Owner SHANGHAI SIMPLEGENE CLINICAL LAB CO LTD
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