PtK2 cells were grown on gold grids and treated with Triton X-100 in a microtubule stabilizing buffer. The resulting cytoskeletons were fixed with glutaraldehyde and subjected to the indirect immunofluorescence procedure using monospecific tubulin antibodies. Grids were examined first by fluorescence microscopy, and the display of fluorescent cytoplasmic microtubules was recorded. The grids were then stained with uranyl acetate and the display of fibrous structures recorded by electron microscopy. Thus the display of cytoplasmic microtubular structures in the light microscope and the electron microscope can be compared within the same cytoskeleton. The results show a direct correspondence of the fluorescent fibers in the light microscope with uninterrupted fibers of diameter approximately 550 A in the electron microscope. This is the diameter reported for a single microtubule decorated around its circumference by two layers of antibody molecules. Thus under optimal conditions immunofluorescence microscopy can visualize individual microtubules.
Article| June 01 1978
Individual microtubules viewed by immunofluorescence and electron microscopy in the same PtK2 cell
Online Issn: 1540-8140
Print Issn: 0021-9525
J Cell Biol (1978) 77 (3): R27.
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M Osborn, RE Webster, K Weber; Individual microtubules viewed by immunofluorescence and electron microscopy in the same PtK2 cell. J Cell Biol 1 June 1978; 77 (3): R27–34. doi: https://doi.org/10.1083/jcb.77.3.R27
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