Fixation strategies for retinal immunohistochemistry.
Fixation strategies for retinal immunohistochemistry.
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DOI：
10.1016/j.preteyeres.2015.04.001
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发表时间：
2015-09
影响因子：
17.8
通讯作者：
Ishida AT
中科院分区：
文献类型：
--
作者：
Stradleigh TW;Ishida AT
免疫组织化学和体外解剖学研究为脊椎动物视网膜神经元的种类、分布和信号成分提供了许多线索。迄今为止发表的大量图像的美丽之处，以及它们提供的定性和定量信息，表明这些方法从根本上是有用的。然而，获得这些图像需要组织处理和暴露在化学溶液中，这些化学溶液在组成、温度和渗透压方面与正常的细胞外液不同。由于差异很大，足以改变神经元的细胞间和细胞内信号，而且视网膜容易受到挤压、剪切和磨损，因此很自然地会怀疑免疫组织化学和解剖学方法是否会干扰或破坏它们旨在检查的细胞。组织固定通常是为了防止这种损害，因此对采集数据的质量和意义至关重要。在这里，我们描述了固定的机制；在免疫组织化学中使用甲醛和戊二醛作为固定剂的优缺点；以及最近被发现改善细胞形状保持和免疫染色模式的广泛使用的方案的修改，特别是在视网膜近端神经元。
Immunohistochemical and ex vivo anatomical studies have provided many glimpses of the variety, distribution, and signaling components of vertebrate retinal neurons. The beauty of numerous images published to date, and the qualitative and quantitative information they provide, indicate that these approaches are fundamentally useful. However, obtaining these images entailed tissue handling and exposure to chemical solutions that differ from normal extracellular fluid in composition, temperature, and osmolarity. Because the differences are large enough to alter intercellular and intracellular signaling in neurons, and because retinae are susceptible to crush, shear, and fray, it is natural to wonder if immunohistochemical and anatomical methods disturb or damage the cells they are designed to examine. Tissue fixation is typically incorporated to guard against this damage and is therefore critically important to the quality and significance of the harvested data. Here, we describe mechanisms of fixation; advantages and disadvantages of using formaldehyde and glutaraldehyde as fixatives during immunohistochemistry; and modifications of widely used protocols that have recently been found to improve cell shape preservation and immunostaining patterns, especially in proximal retinal neurons.
脂质消耗的杆光感受器膜的超微结构。
DOI：
10.1083/jcb.63.2.587
发表时间：
1974-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Nir I;Hall MO
通讯作者：
Hall MO
关于种子的研究。 I.种子的固定。
影响因子：
7.8
作者：
Mollenhauer, H H;Totten, C
通讯作者：
Totten, C