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Published January 15, 2014 | Published
Journal Article Open

Photobleaching imprinting microscopy: seeing clearer and deeper


We present a generic sub-diffraction-limited imaging method – photobleaching imprinting microscopy (PIM) – for biological fluorescence imaging. A lateral resolution of 110 nm was measured, more than a twofold improvement over the optical diffraction limit. Unlike other super-resolution imaging techniques, PIM does not require complicated illumination modules or specific fluorescent dyes. PIM is expected to facilitate the conversion of super-resolution imaging into a routine lab tool, making it accessible to a much broader biological research community. Moreover, we show that PIM can increase the image contrast of biological tissue, effectively extending the fundamental depth limit of multi-photon fluorescence microscopy.

Additional Information

© 2014 The Company of Biologists Ltd. Received 17 September 2013; Accepted 30 October 2013. We thank J. Ballard for close reading of the manuscript. We would also like to thank L. D. Wang and J. Yao for helpful discussions. Competing interests: L.W. has a financial interest in Microphotoacoustics, Inc. and Endra, Inc., which, however, did not support this work. Author contributions: L.G. designed the biological experiment, performed part of the microscopy experiments, analyzed the data, and prepared the manuscript. A.G. analyzed part of data, and prepared the manuscript. Y.L. contributed to experimental design and performed part of microscopy experiments. C.L. prepared the biological sample and performed part of the microscopy experiments. L.V.W. contributed to the conceptual system and experimental design and manuscript preparation. This work was sponsored in part by National Institutes of Health (NIH) [grant numbers DP1 EB016986 (NIH Director's Pioneer Award), R01 EB008085, R01 CA134539, U54 CA136398, R01 CA157277 and R01 CA159959]. Deposited in PMC for release after 12 months.

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