Advanced Microscopy: Laser Scanning Confocal Microscopy
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Fluorescence microscopy is an important and fundamental tool for biomedical research. Optical microscopy is almost non-invasive and allows highly spatially resolved images of organisms, cells, macromolecular complexes, and biomolecules to be obtained. Generally speaking, the architecture of the observed structures is not significantly modified and the environmental conditions can be kept very close to physiological reality. The development of fluorescence microscopy was revolutionized with the invention of laser scanning confocal microscopy (LSCM). With its unique three-dimensional representation and analysis capabilities, this technology gives us a more real view of the world.
This chapter introduces the reader to the methodology of setting up basic experiments for use with a laser scanning confocal microscope. There are practical guidelines about sample preparation for both fixed and living specimens, as well as examples of some of the applications of confocal microscopy.
Key wordsConfocal microscopy Fluorescence microscopy Imaging Organelle markers
- 3.Palsson-McDermott, E. M., Doyle, S. L., McGettrick, A. F., Hardy, M., Husebye, H., Banahan, K., Gong, M., Golenbock, D., Espevik, T., and O’Neill, L. A. (2009) TAG, a splice variant of the adaptor TRAM, negatively regulates the adaptor MyD88-independent TLR4 pathway, Nat Immunol 10, 579–586.PubMedCrossRefGoogle Scholar
- 4.Hanrahan, O., Webb, H., O’Byrne, R., Brabazon, E., Treumann, A., Sunter, J. D., Carrington, M., and Voorheis, H. P. (2009) The glycosylphosphatidylinositol-PLC in Trypanosoma brucei forms a linear array on the exterior of the flagellar membrane before and after activation, PLoS Pathog 5, e1000468.PubMedCrossRefGoogle Scholar
- 8.Harris, J., Schwinn, N., Mahoney, J. A., Lin, H. H., Shaw, M., Howard, C. J., da Silva, R. P., and Gordon, S. (2006) A vitellogenic-like carboxypeptidase expressed by human macrophages is localized in endoplasmic reticulum and membrane ruffles, Int J Exp Pathol 87, 29–39.PubMedCrossRefGoogle Scholar