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  • Tissue sectioning for epifluorescence microscopy

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    69763_1.pdf (1.535Mb)
    Author(s)
    Nguyen, Maria
    Cavanagh, Brenton
    Goodison, Tavia
    Norazit, Anwar
    Meedeniya, Adrian
    Griffith University Author(s)
    Cavanagh, Brenton L.
    Nguyen, Maria T.
    Goodison, Tavia
    Meedeniya, Adrian C.
    Norazit, Anwar
    Year published
    2010
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    Abstract
    Specimens labelled with multiple fluorophores illuminating a variety of cellular targets, facilitated by developments in probes, data acquisition and analysis technologies, have extended the capabilities of anatomical investigations. However, the field has largely neglected development and consensus in optimal techniques of specimen preparation. The optical properties of the specimen largely determines study outcome in epifluorescence microscopy. Thus, we discuss tissue preparation protocols and their application in multiple anatomical investigations. The processing of tissues optimised for multiple studies are detailed. ...
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    Specimens labelled with multiple fluorophores illuminating a variety of cellular targets, facilitated by developments in probes, data acquisition and analysis technologies, have extended the capabilities of anatomical investigations. However, the field has largely neglected development and consensus in optimal techniques of specimen preparation. The optical properties of the specimen largely determines study outcome in epifluorescence microscopy. Thus, we discuss tissue preparation protocols and their application in multiple anatomical investigations. The processing of tissues optimised for multiple studies are detailed. The conservation of cyto-architecture and antigenicity, reduced tissue autofluorescence, increased permeabilisation, and applications for thick tissue sections are examined. Multiple analytical assays of the same tissue sections for the cross correlation of multiple data sets are presented. The application of multiple labelling immunofluorescence, fluorescent in situ hybridisation and fluorescent histochemistry, in qualitative and quantitative analysis are considered. Keywords Anatomy; Frozen sections; Fluorescence histochemistry; Immunofluorescence; in situ hybridisation; Microscopy; Morphometry; Paraffin; Polyethylene glycol; Stereology; Tissue sectioning.
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    Book Title
    Microscopy: Science, Technology, Applications and Education
    Volume
    2
    Publisher URI
    https://www.formatex.org/
    Copyright Statement
    © 2010 Formatex. The attached file is reproduced here in accordance with the copyright policy of the publisher. Please refer to the publisher’s website for further information.
    Subject
    Central Nervous System
    Publication URI
    http://hdl.handle.net/10072/38361
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    • Book chapters

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