Modulated polarization microscopy: a promising new approach to visualizing cytoskeletal dynamics in living cells.
In an effort to visualize cytoskeletal filaments in living cells, we have developed modulated polarization microscopy. Modulated polarization microscopy visualizes cytoskeletal filaments based on their birefringence but differs from the standard polarization microscopy by exploiting the angle dependence of birefringence. A prototype instrument has been developed using two Faraday rotators under computer control to change the angle of plane polarized light at a known rate. By placing one Faraday rotator before and one after the specimen, rotation produced by the first Faraday rotator is cancelled by the second. This allows the use of fixed polarizer and analyzer in a crossed configuration and continuous imaging of the specimen between crossed polarizers. The variation in polarization angle of light illuminating the specimen causes birefringent elements to oscillate in brightness. Images acquired as polarization angle is varied are then processed by a Fourier filter image-processing algorithm. The Fourier filtering algorithm isolates those signals that vary at the proper rate, whereas static or random signals are removed. Here we show that the modulated polarization microscope can reveal cytoskeletal elements including stress fibers and microtubules in living cells.
Kuhn
J R
JR
Division of Molecular Cell and Developmental Biology, University of Texas, Austin, Texas 78712, USA.
Wu
Z
Z
Poenie
M
M
D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D001718:Birefringence; D002460:Cell Line; D003599:Cytoskeleton / Q000737:chemistry* / Q000648:ultrastructure*; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics; D051379:Mice; D008859:Microscopy, Polarization / Q000295:instrumentation / Q000379:methods*; D008870:Microtubules / Q000737:chemistry / Q000648:ultrastructure; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000737:chemistry / Q000648:ultrastructure; D014162:Transfection; D014404:Tubulin / Q000737:chemistry / Q000235:genetics
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D014404:Tubulin; D049452:Green Fluorescent Proteins