Abstract
Correlative Light Electron Microscopy (CLEM) combines advantages of light microscopy and electron microscopy in one experiment to deliver information above and beyond the capability of either modality alone. There are many different CLEM techniques, each having its own special advantages but also its technical challenges. It is however the biological question that (should) drive(s) the development and application of a specific CLEM technique in order to provide the answer. Here we describe the development of a CLEM technique that is based on the Tokuyasu cryo immuno-gold labelling technique that has allowed us to quantitatively study GLUT4 trafficking.
Highlights
glucose transporter type 4 (GLUT4) AbbreviationsGLUT4 Glucose transporter type 4SNARE SNAP receptorVAMP2 vesicle associated membrane protein 2insulin-regulated amino peptidase (IRAP) Interleukin-1 receptor antagonist protein HAInfluenza virus hemagglutininHandling Editor: David RobinsonL
Correlative Light Electron Microscopy (CLEM) is able to deal with very low transfection efficiencies very well in order to obtain quantitative data higher transfection levels were for this particular application desirable
They showed that both differentiated adipocytes and undifferentiated preadipocytes can be successfully transduced with lentiviral vectors, resulting in a roughly 95 % transfection efficiency and no significant toxicity effects, demonstrating a much more suitable method of gene transfer in the 3T3-L1 model system (Carlotti et al 2004)
Summary
The aims of this paper are twofold:
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