Application of photoconversion technique for correlated confocal and ultrastructural studies in organotypic slice cultures
Photoconversion of fluorescent staining into stable diaminobenzidine (DAB) precipitate is widely used for neuroanatomical and developmental studies. An important advantage of the approach is to make correlations between light and electron microscopy analyses possible, the DAB reaction product formed...
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Published in | Microscopy research and technique Vol. 68; no. 2; pp. 90 - 96 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
Hoboken
Wiley Subscription Services, Inc., A Wiley Company
01.10.2005
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Subjects | |
Online Access | Get full text |
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Summary: | Photoconversion of fluorescent staining into stable diaminobenzidine (DAB) precipitate is widely used for neuroanatomical and developmental studies. An important advantage of the approach is to make correlations between light and electron microscopy analyses possible, the DAB reaction product formed during photoconversion being electron dense. By combining a photoconversion approach with biolistic transfection of neurons in organotypic hippocampal slice cultures, we describe here a methodology that allowed us to study at the electron microscopy level the fine details of cells expressing specific genes of interest. The same approach has also been used to analyze the ultrastructural characteristics of specific cells such as neurons recorded with patch clamp techniques. This approach revealed particularly useful for studies of dendritic arborisation, dendritic spines, and axon varicosities of identified cells, as precise morphometric parameters of these structures can only be obtained by electron microscopy. The techniques used for fluorescent staining and photoconversion of these different cell structures and the results obtained by electron microscopic analyses are described. Microsc. Res. Tech. 68:90–96, 2005. © 2005 Wiley‐Liss, Inc. |
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Bibliography: | istex:1C2F222499CCB3035E78D5F4C328A0914BB3E566 ark:/67375/WNG-5D3HZJ3J-6 ArticleID:JEMT20239 Leenaards Foundation Award - No. 1907/ep Novartis Foundation Swiss National Science Foundation - No. 3100A0-105721 ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2 |
ISSN: | 1059-910X 1097-0029 |
DOI: | 10.1002/jemt.20239 |