使用说明：
1. 对于贴壁细胞：
a. 对于6孔板中培养的细胞，吸除细胞培养液，用PBS或HBSS洗涤1次，加入1毫升β-半乳糖苷酶染色固定液，室温固定15分钟。对于其它类型的培养板，固定液及后续溶液的用量参照此比例进行操作。
b. 吸除细胞固定液，用PBS或HBSS洗涤细胞3次，每次3分钟。
c. 吸除PBS或HBSS，每孔加入1毫升染色工作液。使用聚丙烯(polypropylene)容器，不能使用聚苯乙烯(polystyrene)容器配制染色工作液。染色工作液的配制方法参考表1。

说明：对于聚丙烯容器和聚苯乙烯容器的简单的判定方法是：聚丙烯容器可以高温高压灭菌； 而聚苯乙烯容器不适合高温高压灭菌，一旦高温高压处理就会严重变形。配制染色工作液时，可能有少量絮状沉淀出现，震荡混匀后就会完全溶解，且需要待溶解完全后才能使用。
d. 37℃孵育过夜，可以用parafilm或保鲜膜封住6孔板防止蒸发。注意：37℃孵育不能在二氧化碳培养箱中进行。
e. 普通光学显微镜下观察。如不能及时观察计数，可以去除染色工作液，加入2毫升PBS，4℃可以保存数天；或者加上封片液封片后，4℃可以保存较长时间。
2. 对于悬浮细胞：
a. 离心收集细胞至1.5ml离心管内，用PBS或HBSS洗涤1次，加入1毫升β-半乳糖苷酶染色固定液，室温固定15分钟。固定时可以在摇床上缓慢摇动，以避免细胞结成团块。
b. 离心，吸除细胞固定液，用PBS或HBSS洗涤细胞3次，每次3分钟。
c. 离心，吸除PBS或HBSS，每管加入0.5-1毫升染色工作液。染色工作液的配制方法参考表1。
d. 37℃孵育过夜。注意：37℃孵育不能在二氧化碳培养箱中进行。
e. 取部分染色后的细胞，滴加到载玻片上或6孔板内，普通光学显微镜下观察。如不能及时观察计数，可以离心，去除染色工作液，然后加入1毫升PBS，4℃可以保存数天。如果离心，取细胞用于涂片，加上封片液封片后，4℃可以保存较长时间。
3. 对于组织切片：
a. 对于石蜡切片先按照常规方法进行脱蜡和水化处理。对于冷冻切片直接按照以下步骤进行。
b. 加入适当体积的β-半乳糖苷酶染色固定液，以充分盖住组织为宜，室温固定不少于15分钟。
c. 用PBS浸泡洗涤组织3次，每次不少于5分钟。
d. 吸除PBS，加入适当量的染色工作液。染色工作液的配制方法参考表1。
e. 37℃孵育过夜，可以用parafilm或保鲜膜封住防止蒸发。最好把整个切片浸泡在染色工作液中。
注意：37℃孵育不能在二氧化碳培养箱中进行。
f. 普通光学显微镜下观察。如不能及时观察，加上封片液封片后4℃可以保存较长时间。

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