An efficient and reproducible process for transmission electron microscopy (TEM) of rare cell populations.
An efficient and reproducible process for transmission electron microscopy (TEM) of rare cell populations.
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DOI：
10.1016/j.jim.2013.11.025
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发表时间：
2014-02
影响因子：
2.2
通讯作者：
Filippi, Marie-Dominique
中科院分区：
文献类型：
--
作者：
Kumar, Sachin;Ciraolo, Georgianne;Hinge, Ashwini;Filippi, Marie-Dominique
透射电子显微镜 (TEM) 可提供亚细胞器水平的细胞超微结构细节。然而，细胞超微结构的细节以及稀有细胞群中各种细胞器的细胞组织和含量，特别是在悬浮液中，如造血干细胞（HSC）仍然难以捉摸。这主要是由于 TEM 研究需要数百万个细胞。因此，迫切需要一种方法，能够对此类稀有细胞群的低细胞数量进行 TEM 研究。我们描述了一种针对稀有细胞群进行 TEM 研究的替代且新颖的方法。在这里，我们非常轻松地对 10,000 个 HSC 细胞进行了 TEM 研究。特别是，在 PFA-GA 固定后，通过伊文思蓝染色来鉴定微小的细胞沉淀。将细胞沉淀预先包埋在小型微量离心管中的琼脂糖中，并进行脱水、渗透和包埋处理。半薄和超薄切片鉴定出每个切片中有大量细胞簇，高尔基复合体和线粒体的形态和超微结构细节保存完好。总之，该方法提供了一种高效、简单且可重复的过程，可以对有限的生物样品（包括悬浮细胞）进行定性和定量 TEM 分析。
Transmission electron microscopy (TEM) provides ultra-structural details of cells at the sub-organelle level. However, details of the cellular ultrastructure, and the cellular organization and content of various organelles in rare populations, particularly in the suspension, like hematopoietic stem cells (HSCs) remained elusive. This is mainly due to the requirement of millions of cells for TEM studies. Thus, there is a vital requirement of a method that will allow TEM studies with low cell numbers of such rare populations. We describe an alternative and novel approach for TEM studies for rare cell populations. Here we performed TEM study from 10,000 HSC cells with quite ease. In particular, tiny cell pellets were identified by Evans blue staining after PFA-GA fixation. The cell pellet was pre-embedded in agarose in a small microcentrifuge tube and processed for dehydration, infiltration and embedding. Semi-thin and ultra-thin sections identified clusters of numerous cells per sections with well preserved morphology and ultrastructural details of golgi complex and mitochondria. Together, this method provides an efficient, easy and reproducible process to perform qualitative and quantitative TEM analysis from limited biological samples including cells in suspension.
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DOI：
10.1007/bf00310003
发表时间：
1970-01-01
期刊：
HISTOCHEMIE
影响因子：
--
作者：
HOPWOOD, D
通讯作者：
HOPWOOD, D
影响因子：
5.5
作者：
Saini, R.;Patel, S.;Dikshit, M.
通讯作者：
Dikshit, M.
DOI：
10.1016/s0022-5320(65)80075-2
发表时间：
1965-01-01
期刊：
JOURNAL OF ULTRASTRUCTURE RESEARCH
影响因子：
--
作者：
ANDERSON, DR
通讯作者：
ANDERSON, DR
影响因子：
3.4
作者：
MATHER, J;STANBRIDGE, CM;BUTLER, EB
通讯作者：
BUTLER, EB
影响因子：
2.6
作者：
Chen, Jichun;Ellison, Felicia A.;Young, Neal S.
通讯作者：
Young, Neal S.