Title: Visualization of individual astrocytes in three-dimensional cerebellar tissues using green fluorescent protein
Abstract: The central nervous system is a complex structure traditionally studied through selective stains and two-dimensional images of fixed tissues. A long term goal of this laboratory is the study of dynamic cellular interactions in three dimensional tissues such as organotypic cultures. Due to the large volume of tissue, traditional microscopy methods contain excessive out-of-focus data making it difficult to observe clearly specific cellular interactions. The visualization of morphological changes in specific cells in living, three-dimensional tissues is commonly done with vital dyes. An alternate approach is labeling individual astrocytes with Green Fluorescent Protein (GFP) and injecting them into organotypic cultures. GFP is a naturally fluorescent protein originally isolated from the jellyfish Aequorea victorea . To label individual astrocytes, LRM55 astroglial cells were transfected with GFP using calcium phosphate. The percentage of stable transfectants was low (< 1%) resulting in expression of varying amounts of GFP, as seen in Figs 5 and 6.
Publication Year: 1996
Publication Date: 1996-08-11
Language: en
Type: article
Indexed In: ['crossref']
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