Institute of Neurobiology, Biomedical Research Center of the Slovak Academy of Sciences, Šoltésovej 4, Košice 040 01 Slovak Republic.
Institute of Neurobiology, Biomedical Research Center of the Slovak Academy of Sciences, Šoltésovej 4, Košice 040 01 Slovak Republic.
Acta Histochem. 2022 Jan;124(1):151838. doi: 10.1016/j.acthis.2021.151838. Epub 2021 Dec 31.
In this paper, we describe a protocol for a non-penetrating embedding matrix that can be used for frozen or vibratome sectioning of various formaldehyde-fixed tissue specimens. In our experiments, we wanted to prepare thin frozen sections from miniature specimens for fluorescent staining. As we could not achieve satisfactory results with any of the previously published methods, we have tried to modify the existing protocols, and systematically evaluated the effect of these modifications on the properties of the embedding matrix. The resulting protocol is simple, the matrix gets firmly attached to the tissues, does not cause autofluorescence and enables preparing extremely thin frozen sections. The matrix can be used for 1, embedding miniature specimens from problematic tissues to enable cutting very thin frozen sections, 2, grouping multiple specimens into one large block for simultaneous processing, and 3, dispersing single cells and preparing cell blocks for frozen sectioning.
本文描述了一种非穿透性嵌入基质的方案,可用于各种甲醛固定组织标本的冷冻或振动切片。在我们的实验中,我们希望从小型标本中制备薄的冷冻切片用于荧光染色。由于我们无法用任何以前发表的方法获得满意的结果,因此我们试图修改现有的方案,并系统地评估这些修改对嵌入基质性能的影响。所得方案简单,基质牢固地附着于组织,不引起自发荧光,并能够制备极薄的冷冻切片。该基质可用于:1、从小型有问题组织的标本中嵌入,以实现非常薄的冷冻切片的切割;2、将多个标本分组到一个大的块中进行同时处理;3、分散单细胞并制备细胞块进行冷冻切片。