Research Papers: Imaging

Multiphoton microscopy of cleared mouse organs

[+] Author Affiliations
Sonia G. Parra, Thomas H. Chia, Joseph P. Zinter, Michael J. Levene

Yale University, Department of Biomedical Engineering, New Haven, Connecticut 06520

J. Biomed. Opt. 15(3), 036017 (June 21, 2010). doi:10.1117/1.3454391
History: Received March 23, 2010; Revised April 13, 2010; Accepted April 15, 2010; Published June 21, 2010; Online June 21, 2010
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Typical imaging depths with multiphoton microscopy (MPM) are limited to less than 300μm in many tissues due to light scattering. Optical clearing significantly reduces light scattering by replacing water in the organ tissue with a fluid having a similar index of refraction to that of proteins. We demonstrate MPM of intact, fixed, cleared mouse organs with penetration depths and fields of view in excess of 2mm. MPM enables the creation of large 3-D data sets with flexibility in pixel format and ready access to intrinsic fluorescence and second-harmonic generation. We present high-resolution images and 3-D image stacks of the brain, small intestine, large intestine, kidney, lung, and testicle with image sizes as large as 4096×4096pixels.

Figures in this Article
PMTs; HC-125-02, Hamamatsu, Bridgewater, New Jersey

Citation

Sonia G. Parra ; Thomas H. Chia ; Joseph P. Zinter and Michael J. Levene
"Multiphoton microscopy of cleared mouse organs", J. Biomed. Opt. 15(3), 036017 (June 21, 2010). ; http://dx.doi.org/10.1117/1.3454391


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