CN111157319A - 一种革兰氏染色液的脱色液配方及配制方法 - Google Patents
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Abstract
本发明公开了一种革兰氏染色液的脱色液配方,每配制100ml的脱色液中,需要丁酮33‑37ml、无水乙醇63‑67ml。本发明还公开了一种革兰氏染色液的脱色液配制方法,包括以下步骤,步骤1):将丁酮33‑37ml、无水乙醇63‑67ml倒入同一容器,使用搅拌棒对丁酮及无水乙醇搅拌30秒形成脱色液;步骤2):将步骤1)的脱色液倒入磨砂口玻璃瓶中,存放在6—30度的环境中。与现有的多数脱色液相比本革兰氏染色液的脱色液中的丁酮稳定性高、挥发性低,人体吸入量较少,对健康影响较小;其中丁酮的化学性质较稳定,处理方式简单,使操作流程简化,方便了操作,减少了附加成本;同时也保护了环境,减少了有毒气体的排放。
Description
技术领域
本发明涉及革兰氏染色液领域,特别涉及一种革兰氏染色液的脱色液配方及配制方法。
背景技术
革兰氏染色法是细菌学中广泛使用的一种鉴别染色法,这种染色法是由一位丹麦医生汉斯·克里斯蒂安·革兰(Hans Christian Gram,1853年-1938年)于1884年所发明,最初是用来鉴别肺炎球菌与克雷白氏肺炎菌之间的关系。未经染色之细菌,由于其与周围环境折光率差别甚小,故在显微镜下极难观察。染色后细菌与环境形成鲜明对比,可以清楚地观察到细菌的形态、排列及某些结构特征,而用以分类鉴定。染色步骤一般包括初染、媒染、脱色、复染等四个步骤,现今市场上使用的兰氏染色液的脱色液主要是以丙酮与无水乙醇生产制备的。丙酮在脱色液中起到加快脱色的作用。丙酮(acetone,CH3COCH3),又名二甲基酮,为最简单的饱和酮。是一种无色透明液体,有特殊的辛辣气味。易溶于水和甲醇、乙醇、乙醚、氯仿、吡啶等有机溶剂。易燃、易挥发,化学性质较活泼。目前世界上丙酮的工业生产以异丙苯法为主。丙酮在工业上主要作为溶剂用于炸药、塑料、橡胶、纤维、制革、油脂、喷漆等行业中,也可作为合成烯酮、醋酐、碘仿、聚异戊二烯橡胶、甲基丙烯酸甲酯、氯仿、环氧树脂等物质的重要原料。从丙酮的化学性质上来看,属于易挥发性物质,稳定性差。所配置的脱色液在开盖后极容易挥发失效,导致脱色效果变差,实验数据不准确。而且丙酮具有一定的毒性和强挥发性,容易被人体吸入,吸入丙酮会对健康造成极大的影响,操作时要做繁琐的防护工作,对工作区和技术人员水平要求高,操作不方便,增加了后续费用。
发明内容
本发明要解决的技术问题是提供一种革兰氏染色液的脱色液配方及配制方法,该脱色液脱色效果更好、该脱色液的存放时间更长。
为了解决上述技术问题,本发明的技术方案为:
一种革兰氏染色液的脱色液配方,每配制100ml的脱色液中,需要丁酮33-37ml、无水乙醇63-67ml。
进一步地,每配制100ml的脱色液中,需要丁酮35ml、无水乙醇65ml。
进一步地,所述无水乙醇的纯度为95%。
一种革兰氏染色液的脱色液配制方法,包括以下步骤,
步骤1):将丁酮33-37ml、无水乙醇63-67ml倒入同一容器,使用搅拌棒对丁酮及无水乙醇搅拌30秒形成脱色液;
步骤2):将步骤1)的脱色液倒入磨砂口玻璃瓶中,存放在6—30度的环境中。
采用上述技术方案,上述的配方在无水乙醇中配对一定比例丁酮通过上述工艺进行配制,上述比例的丁酮与无水乙醇混合搅拌后形成可以使其溶解力达到比丙酮更优的脱色效果。与现有的多数脱色液相比本革兰氏染色液的脱色液中的丁酮稳定性高、挥发性低,人体吸入量较少,对健康影响较小;其中丁酮的化学性质较稳定,处理方式简单,使操作流程简化,方便了操作,减少了附加成本;同时也保护了环境,减少了有毒气体的排放。
附图说明
图1为本发明实验1两组配制后脱色液静置5分钟的液面状态图;
图2为本发明实验1两组配制后脱色液静置20分钟的液面状态图;
图3为本发明实验1两组配制后脱色液静置1小时20分钟的液面状态图;
图4为本发明实验2通过脱色液A脱色后显微镜成像状态图;
图5为本发明实验2通过脱色液A脱色后载玻片龙胆紫溶液残留的状态图;
图6为本发明实验2通过脱色液B脱色后显微镜成像状态图;
图7为本发明实验2通过脱色液B脱色后载玻片龙胆紫溶液残留的状态图.
具体实施方式
下面结合附图对本发明的具体实施方式作进一步说明。在此需要说明的是,对于这些实施方式的说明用于帮助理解本发明,但并不构成对本发明的限定。此外,下面所描述的本发明各个实施方式中所涉及的技术特征只要彼此之间未构成冲突就可以相互组合。
本发明公开了了一种革兰氏染色液的脱色液配方,每配制100ml的脱色液中,需要丁酮33-37ml、无水乙醇63-67ml。优选地,每配制100ml的脱色液中,需要丁酮35ml、无水乙醇65ml。无水乙醇的纯度为95%。
一种革兰氏染色液的脱色液配制方法,包括以下步骤,
步骤1):将丁酮33-37ml、无水乙醇63-67ml倒入同一容器,使用搅拌棒对丁酮及无水乙醇搅拌30秒形成脱色液;
步骤2):将步骤1)的脱色液倒入磨砂口玻璃瓶中,存放在6—30度的环境中。
上述的配方在无水乙醇中配对一定比例丁酮通过上述工艺进行配制,上述比例的丁酮与无水乙醇混合搅拌后形成可以使其溶解力达到比丙酮更优的脱色效果。与现有的多数脱色液相比本革兰氏染色液的脱色液中的丁酮稳定性高、挥发性低,人体吸入量较少,对健康影响较小;其中丁酮的化学性质较稳定,处理方式简单,使操作流程简化,方便了操作,减少了附加成本;同时也保护了环境,减少了有毒气体的排放。
实验1:
量取贝索品牌的革兰氏染色液的脱色液(下称脱色液A)与本申请的配方配制的革兰氏染色液的脱色液(下称脱色液B)各5ml,倒入有5ml刻度线的试管内,放入试管架中静置。将2款脱色液静置5分钟,然后拍照记录液面(图1),记录完毕后静置15分钟,然后拍照记录第二次液面(图2)。最后静置1小时,记录第三次液面(图3)。整理数据与图片,得出结论。
经过1小时20分钟的常温挥发,两款脱色液的液面有了明显区别:脱色液A液面最低处比脱色液B的液面最低处要更接近5ml刻度线。经过称重测量后,脱色液B的试管内质量比贝索自产的脱色液要多。结合两者的化学性质,发现在同温同压的环境下,丙酮沸点:56.5℃、挥发速度1120、溶解度9.8、极性5.4;丁酮沸点:79.6℃、挥发速度572、溶解度9.2、极性4.5。由试验可知丙酮的挥发性高于丁酮,故丙酮脱色液的稳定性要比丁酮脱色液要差。
实验2:
取用脱色液A及脱色液B,选用大肠埃希菌为试验对象,大肠埃希菌为革兰氏阴性菌,经革兰氏染色法处理后,在显微镜下呈红色丝状。制作两份涂片,进行两个革兰氏染色流程,其中流程1使用脱色液A,流程2使用脱色液B。
试验流程1:
步骤1):涂片固定;
步骤2):加龙胆紫液染1分钟;
步骤3):用纯化水冲洗;
步骤4):加碘溶液覆盖涂面染1分钟;
步骤5):纯化水冲洗,用吸水纸吸去水分;
步骤6):加贝索产脱色液数滴,并轻轻摇动进行脱色,30秒后水洗,吸水纸吸去水分;
步骤7):加沙黄溶液染10秒钟后,纯化水冲洗,等待干燥,进行镜检;
步骤8):油镜观察后拍照记录,并观察涂片染色剂的残留情况,拍照记录。
试验流程2:
步骤1):涂片固定;
步骤2):加龙胆紫液染1分钟;
步骤3):用纯化水冲洗;
步骤4):加碘溶液覆盖涂面染1分钟;
步骤5):纯化水冲洗,用吸水纸吸去水分;
步骤6):加自产脱色液数滴,并轻轻摇动进行脱色,30秒后水洗,吸水纸吸去水分;
步骤7):加沙黄溶液染10秒钟后,纯化水冲洗,等待干燥,进行镜检;
步骤8):油镜观察后拍照记录,并观察涂片染色剂的残留情况,拍照记录。
经过两个试验流程对比,在使用相同的染色液和碘溶液的情况下,通过脱色液A脱色(图4)后成像模糊,观察对象不明显;且载玻片(图5)上有龙胆紫溶液残留,造成实验数据不准确,容易误判。通过脱色液B脱色后(图6)成像清晰,观察对象明显,在显微镜上能清楚的观测到红色的大肠埃希菌;载玻片(图7)上龙胆紫溶液残留少、干净不容易造成误判。
综上所述,从以上两个实验得出,在同温同压的环境下,用丁酮作为脱色溶剂,不易挥发,其开封后的稳定性要比丙酮的强,配制的脱色液保存时间比丙酮的要更长。自产的丁酮脱色液由无水乙醇与丁酮复配,比丙酮脱色液脱色效果更好,降低了实验的误判性。
以上结合附图对本发明的实施方式作了详细说明,但本发明不限于所描述的实施方式。对于本领域的技术人员而言,在不脱离本发明原理和精神的情况下,对这些实施方式进行多种变化、修改、替换和变型,仍落入本发明的保护范围内。
Claims (4)
1.一种革兰氏染色液的脱色液配方,其特征在于:每配制100ml的脱色液中,需要丁酮33-37ml、无水乙醇63-67ml。
2.根据权利要求1所述的革兰氏染色液的脱色液配方,其特征在于:每配制100ml的脱色液中,需要丁酮35ml、无水乙醇65ml。
3.根据权利要求1或2所述的革兰氏染色液的脱色液配方,其特征在于:所述无水乙醇的纯度为95%。
4.一种革兰氏染色液的脱色液配制方法,其特征在于:包括以下步骤,
步骤1):将丁酮33-37ml、无水乙醇63-67ml倒入同一容器,使用搅拌棒对丁酮及无水乙醇搅拌30秒形成脱色液;
步骤2):将步骤1)的脱色液倒入磨砂口玻璃瓶中,存放在6—30度的环境中。
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