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Published October 28, 2013 | Published
Journal Article Open

Optical sectioning by wide-field photobleaching imprinting microscopy


We present a generic wide-field optical sectioning scheme, photobleaching imprinting microscopy (PIM), for depth-resolved cross-sectional fluorescence imaging. Wide-field PIM works by extracting a nonlinear component that depends on the excitation fluence as a result of photobleaching-induced fluorescence decay. Since no specific fluorescent dyes or illumination modules are required, wide-field PIM is easy to implement on a standard microscope. Moreover, wide-field PIM is superior to deconvolution microscopy in removing background fluorescence, yielding a six-fold improvement in image contrast.

Additional Information

© 2013 AIP Publishing LLC. Received 10 September 2013; accepted 8 October 2013; published online 29 October 2013. This work was sponsored in part by National Institutes of Health (NIH) grants DP1 EB016986 (NIH Director's Pioneer Award), R01 EB008085, R01 CA134539, U54 CA136398, R01 CA157277, and R01 CA159959. L.W. has a financial interest in Microphotoacoustics, Inc. and Endra, Inc., which, however, did not support this work.

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