CN110057647A - 非离子型kgm固定病理组织样本的新方法 - Google Patents
非离子型kgm固定病理组织样本的新方法 Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
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Abstract
本发明属医疗领域。本发明使用非离子型KGM(葡甘露聚糖)作为主要成分，以无机盐为辅助成分，进行固定病理组织样本，用以代替高致癌、高刺激性的福尔马林。
Description
技术领域
本发明涉及到使用非离子型KGM(葡甘露聚糖)作为主要成分进行固定病理组织样本的新方法，用以代替高致癌、高刺激性的福尔马林，属医疗领域。
背景技术
病理学诊断，是现代医学的基础性学科。其主要方法，是将离体的新鲜组织样本进行超薄切片，然后在光学显微镜下进行观察，以此来诊断是否发生病变。
离体组织因细胞内的酶的作用，20分钟内就要发生自溶，导致细胞形态被破坏而失去诊断价值。因此，尽快将离体组织进行处理，使其不发生改变，尽可能保持原来的细胞形态，此步骤称之为“固定”，是病理技术的重要基础性措施。
目前，固定组织样本的方法，是使用福尔马林进行浸泡。福尔马林是由甲醛溶液和水按1∶9混合而成。甲醛溶液浓度35％至40％，一般是37％。甲醛的危害极大，主要表现为对皮肤粘膜的刺激作用，可引起眼红、眼痒、咽喉不适或疼痛、声音嘶哑、喷嚏、胸闷、气喘、皮炎等。甲醛有刺激性气味，低浓度即可嗅到。而且，长期、低浓度接触甲醛会引起头痛、头晕、乏力、感觉障碍、免疫力降低，并可出现瞌睡、记忆力减退或神经衰弱、精神抑郁等。甲醛导致的慢性中毒对呼吸系统的危害也是巨大的，长期接触甲醛可引发呼吸功能障碍和肝中毒性病变，表现为肝细胞损伤、肝辐射能异常等。甲醛还有致突变性、致癌性、生殖毒性等。广大病理工作者就是在如此的环境下常年接触甲醛，对身心的危害是极其巨大的。
因此，寻找到有效的、环保的福尔马林替代品来进行组织固定，是十分必要和迫切的。本发明就是要解决上述问题。
发明内容
发明人凭借多年的病理工作经验，并经过大量实践和筛选，找到了可以替代甲醛的物质和方法。
非离子型KGM，中文名称为非离子型葡甘露聚糖，或半乳甘露聚糖，由葡萄糖和甘露糖聚合而形成的杂多糖。常见于植物、酵母等细胞壁中。市售成品主要由甘露聚糖和葡萄糖以β-1，4键键合[摩尔比为1.6∶(1～4)]的高分子量非离子型甘露聚糖(glucomannan)，有少量以β-1，4键的之键结构，沿葡甘露聚糖主链上平均每隔9～19个单糖单位有一个乙酰基，它有助于的溶解。平均分子量20万～200万。其主要来源，是广受大众喜爱的魔芋。因此，该物质具有来源广泛、安全性高等特点。
实际研究表明，非离子型KGM具有很好的防腐保鲜功能。主要原因是由于其特殊的葡萄糖和甘露糖的β-1-4链式结构，它不被细胞内的消化酶所影响，而且自身分子亲水性强，容易渗透至组织内部，进而使细胞内的蛋白产生固化，因此，是进行组织固定的比较理想的基础性物质。经发明人大量实践，发现将非离子型KGM用缓冲液配制成1-3％的溶液，对小体积组织(10cm×10cm以下)的固定效果最好，大体积组织(10cm×10cm以上)，固定效果欠佳。究其原因，是因为非离子型KGM在渗透过程中容易与水分子结合而产生一定的粘度，影响了渗透作用。因此，发明人在溶液里添加了一定种类的无机盐，如氯化钠、氯化钾、碳酸氢钠等，一方面有助于延缓非离子型KGM与水的作用，另一方面也起到增加渗透能力，起到辅助固定的功能。无机盐的添加量，一般在1-5％。浓度过高，会产生组织固缩较大，浓度太低，起不到辅助作用。
因此，本发明的核心内容，就是应用非离子型KGM作为组织固定的主要成分，无机盐作为辅助成分，来对新鲜组织样本进行固定。实际使用效果表明，本发明无色无味，绿色环保，对环境无污染，对人体无害，可以使组织固定的效果接近福尔马林。
实施例
很明显，实施方案不限于实施例。
1)取非离子型KGM 25g，溶解于500ml蒸馏水中。
2)取氯化钠10g，氯化钾2g，氯化钙1g，碳酸氢钠2g溶于另外的500ml蒸馏水中。
3)将上述两种溶液混匀。即为1000ml本发明的固定液。
使用时，直接将小体积新鲜组织样本放入本发明的固定液中，使用量至少是样本体积的5倍.放置24小时后，即可进行脱水等后继操作。
Claims (1)
1.一种非离子型KGM固定病理组织样本的新方法，其特征在于，使用非离子型KGM(葡甘露聚糖)作为主要成分。
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1053089A (zh) * | 1989-12-30 | 1991-07-17 | 中国科学院成都生物研究所 | 一种魔芋多糖固定化微生物细胞和酶的新方法 |
| CN1172529A (zh) * | 1995-02-03 | 1998-02-04 | 竹崎悌二 | 生物检测样品的固定支持方法及其固定支持剂和包埋盒 |
| CN2927032Y (zh) * | 2006-07-20 | 2007-07-25 | 陈锦源 | 病理组织处理装置 |
| CN104399125A (zh) * | 2014-12-01 | 2015-03-11 | 中国人民解放军第三军医大学第三附属医院 | 表皮干细胞向汗腺样上皮细胞分化的方法 |
| CN107376025A (zh) * | 2017-07-16 | 2017-11-24 | 陈强 | 一种用于软骨损伤修复的细胞‑支架复合材料制备方法及应用 |
| CN108503739A (zh) * | 2017-02-24 | 2018-09-07 | 上海蓝帕新材料科技有限公司 | 一种非离子型硬挺剂及其制备方法 |
-
2019
- 2019-04-15 CN CN201910301796.7A patent/CN110057647A/zh active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1053089A (zh) * | 1989-12-30 | 1991-07-17 | 中国科学院成都生物研究所 | 一种魔芋多糖固定化微生物细胞和酶的新方法 |
| CN1172529A (zh) * | 1995-02-03 | 1998-02-04 | 竹崎悌二 | 生物检测样品的固定支持方法及其固定支持剂和包埋盒 |
| CN2927032Y (zh) * | 2006-07-20 | 2007-07-25 | 陈锦源 | 病理组织处理装置 |
| CN104399125A (zh) * | 2014-12-01 | 2015-03-11 | 中国人民解放军第三军医大学第三附属医院 | 表皮干细胞向汗腺样上皮细胞分化的方法 |
| CN108503739A (zh) * | 2017-02-24 | 2018-09-07 | 上海蓝帕新材料科技有限公司 | 一种非离子型硬挺剂及其制备方法 |
| CN107376025A (zh) * | 2017-07-16 | 2017-11-24 | 陈强 | 一种用于软骨损伤修复的细胞‑支架复合材料制备方法及应用 |
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