Integrated fluorescence and transmission electron microscopy.
Correlative microscopy is a powerful technique that combines the strengths of fluorescence microscopy and electron microscopy. The first enables rapid searching for regions of interest in large fields of view while the latter exhibits superior resolution over a narrow field of view. Routine use of correlative microscopy is seriously hampered by the cumbersome and elaborate experimental procedures. This is partly due to the use of two separate microscopes for fluorescence and electron microscopy. Here, an integrated approach to correlative microscopy is presented based on a laser scanning fluorescence microscope integrated in a transmission electron microscope. Using this approach the search for features in the specimen is greatly simplified and the time to carry out the experiment is strongly reduced. The potential of the integrated approach is demonstrated at room temperature on specimens of rat intestine cells labeled with AlexaFluor488 conjugated to wheat germ agglutinin and on rat liver peroxisomes immunolabeled with anti-catalase antibodies and secondary AlexaFluor488 antibodies and 10nm protein A-gold.
Agronskaia
Alexandra V
AV
Molecular Biophysics, Department of Physics, Faculty of Science, Utrecht University, Princetonplein 1, 3584 CC Utrecht, The Netherlands.
Valentijn
Jack A
JA
van Driel
Linda F
LF
Schneijdenberg
Chris T W M
CT
Humbel
Bruno M
BM
van Bergen en Henegouwen
Paul M P
PM
Verkleij
Arie J
AJ
Koster
Abraham J
AJ
Gerritsen
Hans C
HC
D000818:Animals; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D007422:Intestines / Q000166:cytology; D008099:Liver / Q000648:ultrastructure; D018613:Microscopy, Confocal / Q000295:instrumentation* / Q000379:methods; D046529:Microscopy, Electron, Transmission / Q000295:instrumentation* / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods; D020675:Peroxisomes / Q000648:ultrastructure; D051381:Rats
D005456:Fluorescent Dyes