分子生物学实验室中研究对象主要是细胞,DNA,RNA和蛋白质。实验过程中我们常常要对这些实验对象进行观察,提取,裂解等操作。然而,要知道我们人体本身就是由细胞组成的,细胞内包括DNA,RNA,蛋白质。所以对这些实验对象进行操作时,使用的很多试剂也会对我们身体产生危害。今天,小编就为大家介绍一下分子生物学实验室中常见的几种有毒物质以及防护措施。如果有遗漏的,欢迎小伙伴们来信讨论补充。
溴化乙锭EtBr
溴化乙锭(EB)是一种高度灵敏的荧光染色剂,用于观察琼脂糖和聚丙烯酰胺凝胶中的DNA。溴化乙锭用标准302 nm紫外光透射仪激发并放射出橙红色信号。溴化乙锭含有一个可以嵌入DNA堆积碱基之间的一个三环平面基团。它与DNA的结合几乎没有碱基序列特异性。在高离子强度的饱和溶液中,大约每2.5个碱基插入一个溴化乙锭分子。当染料分子插入后,其平面基团与螺旋的轴线垂直并通过范德华力与上下碱基相互作用。这个基团的固定位置及其与碱基的密切接近,导致与DNA结合的染料呈现荧光。如下图所示:
溴化乙锭具有一定的毒性,皮肤吸收可造成伤害。刺激眼睛、皮肤、黏膜和上呼吸道。可诱导突变并可能致癌。戴好手套和护目镜,穿好防护服,在通风橱内小心操作。现在实验室中还有许多溴化乙锭的替代品,比如说GelRed,相对来说比EtBr更安全,想知道EtBr和GelRed的区别,请戳这里。
二甲基亚砜DMSO
二甲基亚砜(DMSO),是一种渗透性保护剂,用途广泛,能够降低细胞冰点,减少冰晶的形成,减轻自由基对细胞损害,改变生物膜对电解质、药物、毒物和代谢产物的通透性。研究表明,DMSO存在严重的毒性作用,与蛋白质疏水集团发生作用,导致蛋白质变性,具有血管毒性和肝肾毒性。
- 吸入:高挥发浓度可能导致头痛,晕眩和镇静。
- 皮肤:能够灼伤皮肤并使皮肤有刺痛感,如同所见的皮疹及水泡一样。若二甲基亚砜与含水的皮肤接触会产生热反应。
- 吸收:吸收危险性很低。
用的时候要避免其挥发,要准备1%-5%的氨水备用,皮肤沾上之后要用大量的水洗以及稀氨水洗涤。
丙烯酰胺Acrylamide
聚丙烯酰胺是SDS-PAGE凝胶中的主要成分。未聚合的丙烯酰胺是一种潜在的神经毒素,可通过皮肤吸收。丙烯酰胺的危害主要是引起神经毒性,同时还有生殖、发育毒性。神经毒性作用表现为周围神经退行性变化和脑中涉及学习、记忆和其他认知功能部位的退行性变化,试验还显示丙烯酰胺是一种可能致癌物,职业接触人群的流行病学观察表明,长期低剂量接触丙烯酰胺会出现嗜睡、情绪和记忆改变、幻觉和震颤等症状,伴随末梢神经病如手套样感觉、出汗和肌肉无力。具有累积毒性,不容易排毒。因此在称量丙烯酰胺和亚甲基双酰胺粉末时,戴好手套和面罩,在化学通风橱内操作。聚合的丙烯酰胺是无毒的,但是使用时也应小心,因为其中可能喊有少量未聚合的丙烯酰胺。(下图所示,聚合的丙烯酰胺单元体)
二硫苏糖醇DTT
二硫苏糖醇(DTT),同巯基乙醇一样是很强的还原剂,散发难闻的气味。可用作蛋白质的保护剂,用于防止蛋白质内的半胱氨酸被氧化。也可用于SDS-PAGE电泳时样品的添加剂,用于彻底打破蛋白质中的二硫键。可因吸入、咽下或皮肤吸收而危害健康。当使用固体或高浓度储存液时,戴手套和护目镜,在通风橱中操作。
苯甲基磺酰氟PMSF
这里必须要给一个大大的危险符号,PMSF剧毒,请一定小心使用!!!
苯甲基磺酰氟(PMSF),常用于蛋白质的提取过程中,是胰蛋白酶、胰凝乳蛋白酶的抑制剂,可抑制丝氨酸蛋白酶例如胰蛋白酶(trypsin)和胰凝乳蛋白酶(chymotrypsin),也可抑制半胱氨酸蛋白酶和乙酰胆碱酯酶,是一种高强度毒性的胆碱酯酶抑制剂。它对呼吸道黏膜、眼睛和皮肤有非常大的破坏性。可因吸入、咽下或皮肤吸收而致命。戴合适的手套和安全眼镜,始终在化学通风橱里使用。在接触到的情况下,要立即用大量的水冲洗眼睛或皮肤,已污染的工作服丢弃掉。
二乙基焦碳酸酯DEPC
DEPC即二乙基焦碳酸酯(diethyl pryrocarbonate),可灭活各种蛋白质,是RNA酶的强抑制剂。DEPC是一种潜在的致癌物质,在操作中应尽量在通风的条件下进行,并避免接触皮肤。DEPC毒性并不是很强,但吸入的毒性是最强的,使用时戴口罩。不小心沾到手上注意立即冲洗,RNaseAway TM试剂可以替代DEPC,操作简单,价格低,且无毒性。只需将RNaseAway TM直接倒在玻璃器皿和塑料器皿的表面,浸泡后用水冲洗去除,即可以快速去除器皿表面的RNase,并且不会残留而干扰后续实验。
紫外光
紫外光是实验室中常见的光源。能够用在无菌室进行灭菌,也会用来观察凝胶中的DNA。紫外光可损伤眼视网膜。切勿用裸眼和没有防护装置的紫外光源。在实验室里常用的紫外光源包括手提式紫外灯和紫外透射仪。只能通过吸收有害波长的滤片或安全玻璃片才能观察。紫外线也是诱变剂和致癌的。为使暴露减少到最低限度,确保紫外光源要采用适当防护装置。在紫外光下操作时要戴合适的预防性手套。
甲醇
甲醇常常用于分析试剂或者清除去油剂,有很大的毒性并易挥发。甲醇的毒性对人体的神经系统和血液系统影响最大,它经消化道、呼吸道或皮肤摄入都会产生毒性反应,甲醇蒸气能损害人的呼吸道粘膜和视力。急性中毒症状有:头疼、恶心、胃痛、疲倦、视力模糊以至失明,继而呼吸困难,最终导致呼吸中枢麻痹而死亡。慢性中毒反应为:眩晕、昏睡、头痛、耳鸣、视力减退、消化障碍。使用时要避免吸入其挥发的汽雾。要戴合适的手套和安全眼镜。
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