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Author Sakumoto, Yasuhiko ♦ Ueta, Hisashi ♦ Yuki, Nobuhiro ♦ Matsuno, Kenjiro
Source J-STAGE
Content type Text
Publisher International Society of Histology and Cytology
Language English
Abstract A need for identifying ganglioside-positive cells with neuronal markers prompted us to establish a reliable method for double or triple immunostaining nervous tissues. Perfusion fixation with paraformaldehyde is typically performed for the routine immunostaining of various neuronal markers but is not suitable for immunostaining gangliosides. Acetone fixation of fresh cryosections is frequently used for ganglioside immunodetection; thus, we tested the effect of acetone treatment for unmasking the antigen epitope of gangliosides (acetone etching) on sections of paraformaldehyde-fixed nervous tissue from rats. Acetone etching significantly retrieved ganglioside immunoreactivity while preserving the immunoreactivity of neuronal markers. Various combinations of gangliosides and neuronal markers could be double-stained by the immunoenzyme method or triple-stained by the immunofluorescence method. This new method may provide additional information regarding the relationship between gangliosides and various neuronal markers from routinely paraformaldehyde-fixed nervous tissues, both freshly prepared specimens and those stocked in the laboratory.
ISSN 09149465
Learning Resource Type Article
Publisher Date 2009-01-01
e-ISSN 13491717
Journal Archives of Histology and Cytology(aohc)
Volume Number 72
Issue Number 2
Page Count 14
Starting Page 77
Ending Page 90


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