Skip to main content
×
Home
    • Aa
    • Aa
  • Get access
    Check if you have access via personal or institutional login
  • Cited by 4
  • Cited by
    This article has been cited by the following publications. This list is generated based on data provided by CrossRef.

    Fukushima, Shoichiro Furukawa, Taichi Niioka, Hirohiko Ichimiya, Masayoshi Miyake, Jun Ashida, Masaaki Araki, Tsutomu and Hashimoto, Mamoru 2014. Y2O3:Tm,Yb nanophosphors for correlative upconversion luminescence and cathodoluminescence imaging. Micron, Vol. 67, p. 90.


    Takizawa, Toshihiro and Robinson, John M. 2012. Correlative Light and Electron MIcroscopy.


    ROBINSON, J.M. and TAKIZAWA, T. 2009. Correlative fluorescence and electron microscopy in tissues: immunocytochemistry. Journal of Microscopy, Vol. 235, Issue. 3, p. 259.


    Vicidomini, Giuseppe Gagliani, Maria C. Cortese, Katia Krieger, Jens Buescher, Peter Bianchini, Paolo Boccacci, Patrizia Tacchetti, Carlo and Diaspro, Alberto 2009. A novel approach for correlative light electron microscopy analysis. Microscopy Research and Technique, p. NA.


    ×

Immunolabeling for Correlative Light and Electron Microscopy on Ultrathin Cryosections

  • Irawati K. Kandela (a1), Reiner Bleher (a2) and Ralph M. Albrecht (a1) (a2) (a3)
  • DOI: http://dx.doi.org/10.1017/S1431927608080239
  • Published online: 01 March 2008
Abstract

Correlative labeling permits colocalization of molecular species for observation of the same sample in light (LM) and electron microscopy (EM). Myosin bands in ultrathin cryosections were labeled using both fluorophore conjugated to secondary antibody (IgG) and colloidal gold (cAu) particles conjugated to primary IgG as reporters for LM and transmission electron microscopy (TEM), respectively. This technique allows rapid evaluation of labeling via LM, prior to more time-consuming observations with TEM and also yields two complementary data sets in one labeling procedure. Quenching of the fluorescent signal was inversely related to the distance between fluorophore and cAu particles. The signal from fluorophore conjugated to secondary antibody was inversely proportional to the size of cAu conjugated to primary antibody. Where fluorophore and cAu were bound to the same antibody, the fluorescence signal was nearly completely quenched regardless of fluorophore excitation or emission wavelength and regardless of particle size, 3 nm and larger. Colloidal metal particles conjugated to primary antibody provide high spatial resolution for EM applications. Fluorophore conjugated to secondary antibody provides spatial resolution well within that of conventional fluorescence microscopy. Use of fluorescent secondary antibody moved the fluorophore a sufficient distance from the cAu particles on the primary antibody to limit quenching of fluorescence.

Copyright
Corresponding author
Corresponding author. E-mail: rmalbrec@facstaff.wisc.edu
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Microscopy and Microanalysis
  • ISSN: 1431-9276
  • EISSN: 1435-8115
  • URL: /core/journals/microscopy-and-microanalysis
Please enter your name
Please enter a valid email address
Who would you like to send this to? *
×

Keywords: