A method for fixing and paraffin embedding tissue to retain the natural fluorescence of reporter proteins

Green fluorescent protein (GFP) and its derivatives are routinely employed as surrogate markers for gene expression and lineage tracing in genetically engineered mice. Tissues from these mice are commonly formalin fixed and paraffin embedded (FFPE) for histological studies. However, this results in...

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Veröffentlicht in:BioTechniques. - 1993. - 59(2015), 3 vom: 08. Sept., Seite 153-5
1. Verfasser: Nakagawa, Akifumi (VerfasserIn)
Weitere Verfasser: Alt, Kate Von, Lillemoe, Keith D, Castillo, Carlos Fernández-Del, Warshaw, Andrew L, Liss, Andrew S
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2015
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't FFPE GFP natural fluorescence Fluorescent Dyes Luminescent Proteins Recombinant Proteins Green Fluorescent Proteins 147336-22-9
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520 |a Green fluorescent protein (GFP) and its derivatives are routinely employed as surrogate markers for gene expression and lineage tracing in genetically engineered mice. Tissues from these mice are commonly formalin fixed and paraffin embedded (FFPE) for histological studies. However, this results in inactivation of the natural fluorescence of these proteins, requiring their detection by immunological techniques. Here we present an ethanol fixation protocol that allows for the direct visualization of the natural fluorescence of reporter proteins while maintaining excellent tissue histology. We demonstrate the utility of this method for visualizing green and red fluorescent proteins in a wide range of murine tissues using both cytoplasmic and membrane-localized fluorescent reporter proteins. Tissues fixed by this method also allow for immunohistochemical studies, providing a single method to visualize the natural fluorescence of reporter proteins with subsequent detection of cellular proteins 
650 4 |a Journal Article 
650 4 |a Research Support, N.I.H., Extramural 
650 4 |a Research Support, Non-U.S. Gov't 
650 4 |a FFPE 
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650 4 |a natural fluorescence 
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700 1 |a Alt, Kate Von  |e verfasserin  |4 aut 
700 1 |a Lillemoe, Keith D  |e verfasserin  |4 aut 
700 1 |a Castillo, Carlos Fernández-Del  |e verfasserin  |4 aut 
700 1 |a Warshaw, Andrew L  |e verfasserin  |4 aut 
700 1 |a Liss, Andrew S  |e verfasserin  |4 aut 
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