Green fluorescent protein as a tracer in chimeric tissues: the power of vapor fixation

Jockusch H, Eberhard D (2007)
Methods Mol Biol 411: 145-154.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Autor*in
Jockusch, HaraldUniBi; Eberhard, Daniel
Abstract / Bemerkung
Green fluorescent protein (GFP) and its variants, small, highly soluble proteins, are routinely used as reporters for patterns of gene expression and the origin of cells in transplantation experiments. When not linked as fusion proteins to other polypeptides, they distribute rapidly in the cytoplasm of a given cell, thus allowing real-time observations on living material. For histological analysis, previous bath fixation of whole organs or tissues seemed obligatory, because, during drop fixation of sections, GFP rapidly leaks from cells whose membrane has been damaged by freezing and/or sectioning. The fluorescence of GFP and its derivatives is retained upon fixation, but most enzyme and antigenic activities of interest will be lost in the whole sample as a consequence of formaldehyde (FA) fixation. We have therefore developed an alternative method to fix GFP in frozen tissue sections by FA vapor. This method prevents leakage and redistribution of GFP and allows any cytochemical method to be applied to unfixed adjacent serial sections.
Erscheinungsjahr
2007
Zeitschriftentitel
Methods Mol Biol
Band
411
Seite(n)
145-154
ISBN
9781588297396
ISSN
1064-3745
Page URI
https://pub.uni-bielefeld.de/record/1946058

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Jockusch H, Eberhard D. Green fluorescent protein as a tracer in chimeric tissues: the power of vapor fixation. Methods Mol Biol. 2007;411:145-154.
Jockusch, H., & Eberhard, D. (2007). Green fluorescent protein as a tracer in chimeric tissues: the power of vapor fixation. Methods Mol Biol, 411, 145-154. https://doi.org/10.1007/978-1-59745-549-7_11
Jockusch, Harald, and Eberhard, Daniel. 2007. “Green fluorescent protein as a tracer in chimeric tissues: the power of vapor fixation”. Methods Mol Biol 411: 145-154.
Jockusch, H., and Eberhard, D. (2007). Green fluorescent protein as a tracer in chimeric tissues: the power of vapor fixation. Methods Mol Biol 411, 145-154.
Jockusch, H., & Eberhard, D., 2007. Green fluorescent protein as a tracer in chimeric tissues: the power of vapor fixation. Methods Mol Biol, 411, p 145-154.
H. Jockusch and D. Eberhard, “Green fluorescent protein as a tracer in chimeric tissues: the power of vapor fixation”, Methods Mol Biol, vol. 411, 2007, pp. 145-154.
Jockusch, H., Eberhard, D.: Green fluorescent protein as a tracer in chimeric tissues: the power of vapor fixation. Methods Mol Biol. 411, 145-154 (2007).
Jockusch, Harald, and Eberhard, Daniel. “Green fluorescent protein as a tracer in chimeric tissues: the power of vapor fixation”. Methods Mol Biol 411 (2007): 145-154.

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Optimized protocol for immunostaining of experimental GFP-expressing and human hearts.
Zaglia T, Di Bona A, Chioato T, Basso C, Ausoni S, Mongillo M., Histochem Cell Biol 146(4), 2016
PMID: 27311322
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