Authors: HILL, D. B.; MACOSKO, J. C.; HOLZWARTH, G. M.

Source: Journal of Microscopy, Volume 231, Number 3, September 2008 , pp. 433-439(7)

Publisher: Wiley-Blackwell

Abstract:

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.

Keywords: Background subtraction; DIC; image enhancement; organelle transport; VE-DIC; vesicle tracking

Document Type: Research article

DOI: http://dx.doi.org/10.1111/j.1365-2818.2008.02054.x

Affiliations: 1: Department of Physics, Wake Forest University, Winston-Salem, NC 27109, U.S.A.

Publication date: 2008-09-01

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