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Motion‐enhanced, differential interference contrast (MEDIC) microscopy of moving vesicles in live cells: VE‐DIC updated
Author(s) -
HILL D. B.,
MACOSKO J. C.,
HOLZWARTH G. M.
Publication year - 2008
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.2008.02054.x
Subject(s) - differential interference contrast microscopy , contrast (vision) , computer vision , background subtraction , subtraction , artificial intelligence , differential (mechanical device) , vesicle , interference (communication) , image subtraction , optics , video microscopy , microscopy , computer science , physics , image processing , materials science , pixel , image (mathematics) , chemistry , mathematics , channel (broadcasting) , biology , telecommunications , microbiology and biotechnology , binary image , membrane , arithmetic , biochemistry , thermodynamics
Summary Video‐enhanced differential interference contrast microscopy with background subtraction has made visible many structures and processes in living cells. In video‐enhanced differential interference contrast, the background image is stored manually by defocusing the microscope before images are acquired. We have updated and improved video‐enhanced differential interference contrast by adding automatic generation of the background image as a rolling average of the incoming image stream. Subtraction of this continuously updated 12‐bit background image from the incoming 12‐bit image stream provides a flat background which allows the contrast of moving objects, such as vesicles, to be strongly enhanced while suppressing stationary features such as the overall cell shape. We call our method MEDIC, for motion‐enhanced differential interference contrast. By carrying out background subtraction with 12‐bit images, the number of grey levels in the moving vesicles can be maximized and a single look‐up table can be applied to the entire image, enhancing the contrast of all vesicles simultaneously. Contrast is increased by as much as a factor of 13. The method is illustrated with raw, background and motion‐enhanced differential interference contrast images of moving vesicles within a neurite of a live PC12 cell and a live chick motorneuron.