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标题: 求助 HE染色,细胞边界不清晰,胞核没有着色! [打印本页]

作者: tongttqq    时间: 2006-8-22 12:11     标题: 求助 HE染色,细胞边界不清晰,胞核没有着色!

我是新手,第一次染色居然遇到这种问题。在版内看过,没有同类问题的,还请大家指教!
做的是肝组织的HE染色,在反篮那步就看不清细胞轮廓了。全部完成后,片子还是内细胞边界不清晰,胞核没有着色。这是怎么回事咧?
还有,出现这种状况和当初固定时间是否有关系?
谢谢了!
作者: dxx029    时间: 2006-8-24 10:25

可能的情况是 取材固定的过程中出现问题,是否是取材过程费时长,组织出现自溶现象
                                                                  固定时间不足或是固定时间过长,导致组织的结构不好
作者: tongttqq    时间: 2006-8-29 16:03

组织自溶是什么意思啊,只是看资料说可能是啊,固定时间过长,可能引起PH过酸的,所以染色之前需要浸泡在碳酸锂溶液中,是这样子么?
作者: 刘小玲    时间: 2006-10-11 19:35     标题: 回复 #3 tongttqq 的帖子

烤片的温度较高时间长较长,细胞核受热熟了,所以核不着色,细胞结构不清。
作者: hunter3035    时间: 2006-10-24 20:47

呵呵脱蜡不彻底.
作者: folksmile    时间: 2006-10-26 11:47     标题: Copy from Protocol Online

Hematoxylin and Eosin (H&E) staining
lace slides containing paraffin sections in a slide holder (glass or metal)
•Deparaffinize and rehydrate sections:
3 x 3´ Xylene (blot excess xylene before going into ethanol)
3 x 3´ 100% ethanol
1 x 3´ 95% ethanol
1 x 3´ 80% ethanol
1 x 5´ deionized H2O
•While sections are in water, skim surface of hematoxalin with a Kimwipe to
remove oxidized particles. Blot excess water from slide holder before going into
hematoxalin.
•Hematoxalin staining:
1 x 3´ Hematoxalin
Rinse deionized water
1 x 5´ Tap water (to allow stain to develop)
Dip 8-12x (fast) Acid ethanol (to destain)
Rinse 2 x 1´ Tap water
Rinse 1 x 2´ Deionized water (can leave overnight at this stage)
•Blot excess water from slide holder before going into eosin.
•Eosin staining and dehydration:
1 x 30 sec Eosin (up to 45 seconds for an older batch of eosin)
3 x 5´ 95% ethanol
3 x 5´ 100% ethanol (blot excess ethanol before going into xylene)
3 x 15´ Xylene
•You can leave slides in xylene overnight to get good clearing of any water.
•Coverslip slides using Permount (xylene based).
lace a drop of Permount on the slide using a glass rod, taking care to leave
no bubbles.
•Angle the coverslip and let fall gently onto the slide. Allow the Permount to
spread beneath the coverslip, covering all the tissue.
•Dry overnight in the hood.
作者: tongttqq    时间: 2006-11-7 11:21

谢谢啊,我的问题已经解决了!
作者: zhba0918    时间: 2006-12-27 11:20     标题: 回复 #7 tongttqq 的帖子

哥们怎么解决得,我也碰到了类似得问题!分享一下你的经验吧!在此有礼了!




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