Quasi-confocal fluorescence sectioning with dynamic speckle illumination

Cathie Ventalon; Jerome Mertz

doi:10.1117/12.646889

23 February 2006 Quasi-confocal fluorescence sectioning with dynamic speckle illumination

Cathie Ventalon, Jerome Mertz

Proceedings Volume 6090, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII; 60900M (2006) https://doi.org/10.1117/12.646889
Event: SPIE BiOS, 2006, San Jose, California, United States

Abstract

We present a new fluorescence microscopy technique that provides depth discrimination in thick tissue. The technique relies on a simple modification to a conventional wide-field microscope, and consists in illuminating the sample with a sequence of speckle patterns and displaying the RMS or the variance of the resultant sequence of fluorescent images. We demonstrate quasi-confocal optical sectioning with an axial resolution of 5 microns FWHM. The lateral resolution is identical to the widefield microscope, namely 0.6 micron FWHM. Images of a mouse brain are compared with standard wide-field images, demonstrating an efficient quasi-sectioning capacity in thick tissue throughout approximatively 100 microns depth. This is achieved because of the high contrast maintained by speckle in a scattering media.

Citation Download Citation

Cathie Ventalon and Jerome Mertz "Quasi-confocal fluorescence sectioning with dynamic speckle illumination", Proc. SPIE 6090, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII, 60900M (23 February 2006); https://doi.org/10.1117/12.646889

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KEYWORDS

Speckle

Luminescence

Confocal microscopy

Speckle pattern

Microscopes

Tissues

Objectives

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