CN111829858A - 实验发现两种简单、低廉、来源丰富的新型组织切片染色剂 - Google Patents
实验发现两种简单、低廉、来源丰富的新型组织切片染色剂 Download PDFInfo
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- 238000000034 method Methods 0.000 description 7
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 6
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- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
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Abstract
实验发现两种简单、低廉、来源丰富的新型组织切片染色剂。技术领域：两种服装染料紫BL和蓝B2R，首次通过实验被发现具有良好的组织切片染色效果，主要是对小鼠肝脏等组织切片染色清晰有效。要解决的技术问题：通过服装染料对小鼠肝脏组织切片染色结果观察，筛选到对小鼠肝脏等组织切片染色清晰有效的染料。解决该问题的技术方案的要点：分别用27种直接服装染料进行小鼠肝组织切片染色。利用肝脏组织切片染色效果良好的染料紫BL和蓝B2R，再进行心脏等组织切片染色观察。主要用途：比苏木精廉价、配制简单、来源丰富的服装染料紫BL和蓝B2R，可以作为新型组织切片染色剂替代苏木精，对动物肝脏等组织切片进行染色观察。
Description
技术领域
本发明涉及实验发现两种新型组织切片染色剂。具体地说，本发明涉及两种服装染料紫BL和蓝B2R，首次通过实验被发现具有良好的组织切片染色效果，主要是对小鼠肝脏、心脏、肾脏和睾丸组织切片进行染色后，结果清晰有效。
背景技术
目前用于组织切片染色的染料的种类很多，但是制片染色最常用的还是苏木素-伊红染色法，简称HE染色法。
天然染料苏木精是从南美的苏木干枝中用乙醚浸制出来的一种色素，产量有限，价格较高。使用液配制繁琐耗时。Harris苏木素液配置步骤繁琐，需要半个月以上。Ehrlich苏木素液配方更繁琐，约二个月后才可使用。
可见有必要通过实验寻找操作简单，价格低廉，来源丰富的染色剂。因为目前国内外还没有常用服装染料用于组织切片染色的实验报道，所以，本研究中应用常用服装染料进行组织切片染色实验。
发明内容
首次用服装染料紫BL和蓝B2R对小鼠肝脏、心脏、肾脏和睾丸组织切片进行染色效果观察。结果为组织结构染色清晰，类似于H-E染色结果。这两种染料的组织切片染色清晰有效，比苏木精廉价、简单、来源丰富，可能替代HE染色法中的苏木精。
实验所用各种直接服装染料以蒸馏水溶解，配制浓度为1g/50ml，溶液颜色都较深。
染色步骤基本按照HE染色步骤进行操作。修改两处，一是将苏木素染液替换为直接服装
染料；二是去除或不去除伊红染色这一步。
分别用这27种直接服装染料进行小鼠肝组织切片染色，镜下观察染色效果，进行初筛。经过初筛发现紫BL和蓝B2R的小鼠肝组织切片染色，都比较清晰，能够看清细胞核等HE染色看到的组织结构。
经过对照实验，发现染色效果比较好的染色浓度为1g/L，染色时间为5min。
将染色比较好的紫BL和蓝B2R做进一步的研究，进行小鼠心脏，肾脏和睾丸组织的染色实验，染色结果为紫色或蓝色，结构清晰。肝切片中的中央静脉、肝血窦、肝细胞核、内皮细胞核等结构清晰；心脏纵切片中血管、血细胞、心肌细胞核、内皮细胞核、横纹结构等清晰可辨；肾脏切片中血管球、肾小囊腔、肾小管等结构清晰；睾丸组织切片中生精小管的精原细胞、初级精母细胞、次级精母细胞、精子细胞、精子、肌样细胞以及睾丸间质细胞等结构清晰可辨(图1；图2)
应用直接服装染料紫BL和蓝B2R，分别与伊红对肝和肾组织切片进行共同染色，染色结果与H-E染色结果类似，结构清晰。细胞核被染成紫色或蓝色，细胞质及细胞外基质被染成红色并带有一些紫色或蓝色(图3)。
本发明的有益结果是，这两种染料的组织切片染色清晰有效，比苏木精廉价、简单、来源丰富，可能替代HE染色法中的苏木精。
附图说明
图1肝、心、肾和睾丸组织紫BL染料染色结果
L：肝；H：心脏；K：肾脏；T：睾丸
图2肝、心、肾和睾丸组织蓝B2R染料染色结果
L：肝；H：心脏；K：肾脏；T：睾丸
图3肝和肾组织被紫BL和蓝B2R分别与伊红共同染色的结果
HP：小鼠肝脏切片，紫BL和伊红共同染色的结果
KP：小鼠肾脏切片，紫BL伊红共同染色的结果
HB：小鼠肝脏切片，蓝B2R和伊红共同染色的结果
KB：小鼠肾脏切片，蓝B2R和伊红共同染色的结果
具体实施方式
染色步骤基本按照HE染色常规步骤进行操作。修改两处，一是将苏木素染液替换为直接服装染料；二是去除或不去除伊红染色这一步。
Claims (2)
1.两种服装染料紫BL和蓝B2R，首次通过实验被发现具有良好的组织切片染色效果，主要是对小鼠肝脏、心脏、肾脏和睾丸组织切片进行染色，结果清晰有效。
2.根据权利要求1所述的内容，其特征在于两种服装染料紫BL和蓝B2R对小鼠肝脏、心脏、肾脏和睾丸组织切片进行染色后，观察结果为清晰有效。
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4853210A (en) * | 1984-04-27 | 1989-08-01 | Cytocolor, Inc. | Method of staining cells with a diazo dye and compositions thereof |
| JP2004347594A (ja) * | 2003-04-30 | 2004-12-09 | Shimadzu Corp | 生体標本への試薬の分注方法及び生体標本の解析方法 |
| CN103415762A (zh) * | 2011-01-10 | 2013-11-27 | 文塔纳医疗系统公司 | 苏木精染色方法 |
| US20150369708A1 (en) * | 2013-02-05 | 2015-12-24 | Tripath Imaging, Inc. | Cytological staining compositions and uses thereof |
| CN107205980A (zh) * | 2014-10-21 | 2017-09-26 | M·布达克 | 具有获自虞美人的新型分子的组织和细胞染色剂制剂 |
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- 2019-04-18 CN CN201910351942.7A patent/CN111829858A/zh active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4853210A (en) * | 1984-04-27 | 1989-08-01 | Cytocolor, Inc. | Method of staining cells with a diazo dye and compositions thereof |
| JP2004347594A (ja) * | 2003-04-30 | 2004-12-09 | Shimadzu Corp | 生体標本への試薬の分注方法及び生体標本の解析方法 |
| CN103415762A (zh) * | 2011-01-10 | 2013-11-27 | 文塔纳医疗系统公司 | 苏木精染色方法 |
| US20150369708A1 (en) * | 2013-02-05 | 2015-12-24 | Tripath Imaging, Inc. | Cytological staining compositions and uses thereof |
| CN107205980A (zh) * | 2014-10-21 | 2017-09-26 | M·布达克 | 具有获自虞美人的新型分子的组织和细胞染色剂制剂 |
Non-Patent Citations (3)
| Title |
|---|
| 上海市纺织工业局编写组: "《染料应用手册 上册》", 30 April 1994, 中国纺织出版社, pages: 184 - 186 * |
| 何岩彬: "《染料品种大全 上卷》", 30 June 2018, 沈阳出版社, pages: 178 * |
| 来茂德: "《病理学高级教程》", 人民军医出版社, pages: 766 * |
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