CN113218732A - 一种伊红染液及其制备方法 - Google Patents
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Abstract
本发明公开了一种伊红染液及其制备方法,所述伊红染液的成分包括蒸馏水、水溶性伊红Y、冰醋酸、助染剂、防腐剂,各成分的质量百分比为:蒸馏水99.3~99.5%、水溶性伊红Y0.1~0.3%、冰醋酸0.05~0.2%、助染剂0.01~0.1%、防腐剂0.05~0.1%。其制备方法包括以下步骤:S1、按计量称取蒸馏水、水溶性伊红Y、冰醋酸、助染剂、防腐剂;S2、在烧杯中依次加入蒸馏水、水溶性伊红Y、助染剂、防腐剂,搅拌均匀后得到混合溶液;S3、在混合溶液中加入冰醋酸,将混合溶液的PH值调整为5.5~6.5;S4、将调整PH值后的混合溶液进行过滤操作,得到的滤液即为伊红染液。
Description
技术领域
本发明涉及生化领域,尤其涉及一种伊红染液及其制备方法。
背景技术
苏木精—伊红染色法(hematoxylin-eosinstaining),简称HE染色法,其为石蜡切片技术里常用的染色法之一。苏木精染液为碱性,主要使细胞核内的染色质与胞质内的核酸着紫蓝色;伊红为酸性染料,主要使细胞质和细胞外基质中的成分着红色。HE染色法是组织学、胚胎学、病理学教学与科研中最基本、使用最广泛的技术方法。
苏木精—伊红染色法中的操作步骤主要包括切片—烤片—脱蜡—水化—苏木素染色—分化—返蓝—伊红染色—脱水—透明—封片;其中,伊红染色主要用到水溶性伊红Y,水溶性伊红Y,也称曙红钠盐、黄光曙红或四溴荧光素钠,子式为C20H6O5Br4Na2,易溶于水。
伊红最适宜与苏木精配合染色以显示正常或病理组织的形态结构。水溶性伊红Y中含有一个醌型苯环的发色团和两个形成钠盐的酸性助色团(R-COONa、R-ONa)。这种形成盐类的酸性染料在水中溶解时解离为带负电荷的色酸部分(R-COO-、R-O-),即染料的有色部分和带正电荷的钠离子部分(Na+)。染色时伊红Y带负电荷的色酸部分与组织内带正电荷的物质以离子键的形式结合而显色。细胞浆、红细胞、肌肉、结缔组织、嗜伊红颗粒等被染成不同程度的红色或粉红色,与蓝色的细胞核形成鲜明对比。
现有的水溶性伊红染料配方虽然简单,但染色时间长,染液有效期短,常需加入冰醋酸进行配合使用,但加入冰醋酸的剂量不易掌握,加入的量少达不到促染作用,加入过多则导致染液易沉淀,并且会出现色调不正、容易褪色的现象,染色结果不能长期保存。
发明内容
本发明目的是针对上述问题,提供一种提高细胞组织染色效果的水溶性伊红染液及其制备方法。
为了实现上述目的,本发明的技术方案是:
一种伊红染液,所述伊红染液的成分包括蒸馏水、水溶性伊红Y、冰醋酸、助染剂、防腐剂,各成分的质量百分比为:蒸馏水99.3~99.5%、水溶性伊红Y0.1~0.3%、冰醋酸0.05~0.2%、助染剂0.01~0.1%、防腐剂0.05~0.1%。
进一步的,所述伊红染液中各成分的质量百分比为:蒸馏水99.4%、水溶性伊红Y0.2%、冰醋酸0.2%、助染剂0.1%、防腐剂0.1%。
进一步的,所述助染剂为甲基丙烯酸丁酯、聚丙烯酸钠中的一种或两种混合。
进一步的,所述防腐剂为二叔丁基苯酚、3,5-二叔丁基-4-羟基苯甲酸中的一种或两种混合。
一种伊红染液的制备方法,包括以下步骤:
S1、按计量称取蒸馏水、水溶性伊红Y、冰醋酸、助染剂、防腐剂;
S2、在烧杯中依次加入蒸馏水、水溶性伊红Y、助染剂、防腐剂,搅拌均匀后得到混合溶液;
S3、在混合溶液中加入冰醋酸,将混合溶液的PH值调整为5.5~6.5;
S4、将调整PH值后的混合溶液进行过滤操作,得到的滤液即为伊红染液。
进一步的,所述步骤S4中进行过滤时使用1000~2000目的过滤网进行过滤。
与现有技术相比,本发明具有的优点和积极效果是:
本发明中的伊红染液除了含有冰醋酸作促染剂以外,还加入了(特色)助染剂和防腐剂,提高了伊红染液的使用效果;并且本发明的伊红染液具有操作简便、染色性质稳定、着色力强、颜色鲜艳等优点,有效提高了胞浆部位的染色效果,具有较好的市场推广价值。
具体实施方式
下面将结合本发明的实施例,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
实施例1、本实施例公开了一种伊红染液,所述伊红染液的成分包括蒸馏水、水溶性伊红Y、冰醋酸、甲基丙烯酸丁酯、二叔丁基苯酚,各成分的质量百分比为:蒸馏水99.3%、水溶性伊红Y0.3%、冰醋酸0.2%、甲基丙烯酸丁酯0.1%、二叔丁基苯酚0.1%。
上述伊红染液的制备方法,包括以下步骤:
S1、按计量称取蒸馏水、水溶性伊红Y、冰醋酸、甲基丙烯酸丁酯、二叔丁基苯酚;
S2、在烧杯中依次加入蒸馏水、水溶性伊红Y、甲基丙烯酸丁酯、二叔丁基苯酚,搅拌均匀后得到混合溶液;
S3、在混合溶液中加入冰醋酸,将混合溶液的PH值调整为5.5;
S4、使用1000目的过滤网对调整PH值后的混合溶液进行过滤操作,得到的滤液即为伊红染液。
实施例2、本实施例公开了一种伊红染液,所述伊红染液的成分包括蒸馏水、水溶性伊红Y、冰醋酸、聚丙烯酸钠、3,5-二叔丁基-4-羟基苯甲酸,各成分的质量百分比为:蒸馏水99.4%、水溶性伊红Y0.2%、冰醋酸0.2%、聚丙烯酸钠0.1%、3,5-二叔丁基-4-羟基苯甲酸0.1%。
上述伊红染液的制备方法,包括以下步骤:
S1、按计量称取蒸馏水、水溶性伊红Y、冰醋酸、聚丙烯酸钠、3,5-二叔丁基-4-羟基苯甲酸;
S2、在烧杯中依次加入蒸馏水、水溶性伊红Y、聚丙烯酸钠、3,5-二叔丁基-4-羟基苯甲酸,搅拌均匀后得到混合溶液;
S3、在混合溶液中加入冰醋酸,将混合溶液的PH值调整为6;
S4、使用2000目的过滤网对调整PH值后的混合溶液进行过滤操作,得到的滤液即为伊红染液。
实施例3、本实施例公开了一种伊红染液,所述伊红染液的成分包括蒸馏水、水溶性伊红Y、冰醋酸、甲基丙烯酸丁酯、二叔丁基苯酚,各成分的质量百分比为:蒸馏水99.5%、水溶性伊红Y0.25%、冰醋酸0.05%、甲基丙烯酸丁酯0.1%、二叔丁基苯酚0.1%。
上述伊红染液的制备方法,包括以下步骤:
S1、按计量称取蒸馏水、水溶性伊红Y、冰醋酸、甲基丙烯酸丁酯、二叔丁基苯酚;
S2、在烧杯中依次加入蒸馏水、水溶性伊红Y、甲基丙烯酸丁酯、二叔丁基苯酚,搅拌均匀后得到混合溶液;
S3、在混合溶液中加入冰醋酸,将混合溶液的PH值调整为6.5;
S4、使用2000目的过滤网对调整PH值后的混合溶液进行过滤操作,得到的滤液即为伊红染液。
本发明伊红染液中的水溶性伊红Y主要起染色作用,蒸馏水用于溶解水溶性伊红Y,冰醋酸主要作为促染剂;另外通过加入助染剂,可以令染色效果更鲜明;防腐剂可以使溶剂更加稳定,令切片的储存时间更长。
本发明中的伊红染液除了含有冰醋酸作促染剂以外,还加入了(特色)助染剂和防腐剂,提高了伊红染液的使用效果;并且本发明的伊红染液具有操作简便、染色性质稳定、着色力强、颜色鲜艳等优点,有效提高了胞浆部位的染色效果,具有较好的市场推广价值。
Claims (6)
1.一种伊红染液,其特征在于:所述伊红染液的成分包括蒸馏水、水溶性伊红Y、冰醋酸、助染剂、防腐剂,各成分的质量百分比为:蒸馏水99.3~99.5%、水溶性伊红Y 0.1~0.3%、冰醋酸0.05~0.2%、助染剂0.01~0.1%、防腐剂0.05~0.1%。
2.如权利要求1所述的伊红染液,其特征在于:所述伊红染液中各成分的质量百分比为:蒸馏水99.4%、水溶性伊红Y0.2%、冰醋酸0.2%、助染剂0.1%、防腐剂0.1%。
3.如权利要求1或权利要求2任一所述的伊红染液,其特征在于:所述助染剂为甲基丙烯酸丁酯、聚丙烯酸钠中的一种或两种混合。
4.如权利要求3所述的伊红染液,其特征在于:所述防腐剂为二叔丁基苯酚、3,5-二叔丁基-4-羟基苯甲酸中的一种或两种混合。
5.一种如权利要求4所述的伊红染液的制备方法,其特征在于:包括以下步骤:
S1、按计量称取蒸馏水、水溶性伊红Y、冰醋酸、助染剂、防腐剂;
S2、在烧杯中依次加入蒸馏水、水溶性伊红Y、助染剂、防腐剂,搅拌均匀后得到混合溶液;
S3、在混合溶液中加入冰醋酸,将混合溶液的PH值调整为5.5~6.5;
S4、将调整PH值后的混合溶液进行过滤操作,得到的滤液即为伊红染液。
6.如权利要求5所述的伊红染液的制备方法,其特征在于:所述步骤S4中进行过滤时使用1000~2000目的过滤网进行过滤。
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