Wavefront shaping of a Bessel light field enhances light sheet microscopy with scattered light

J. Nylk; C. Mitchell; T. Vettenburg; F. J. Gunn-Moore; K. Dholakia

doi:10.1117/12.2038525

12 March 2014 Wavefront shaping of a Bessel light field enhances light sheet microscopy with scattered light

J. Nylk, C. Mitchell, T. Vettenburg, F. J. Gunn-Moore, K. Dholakia

Proceedings Volume 8949, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXI; 89490V (2014) https://doi.org/10.1117/12.2038525
Event: SPIE BiOS, 2014, San Francisco, California, United States

Abstract

Light sheet microscopy has seen a resurgence as it facilitates rapid, high contrast, volumetric imaging with minimal sample exposure. Initially developed for imaging scattered light, this application of light sheet microscopy has largely been overlooked but provides an endogenous contrast mechanism which can complement fluorescence imaging and requires very little or no modification to an existing light sheet fluorescence microscope. Fluorescence imaging and scattered light imaging differ in terms of image formation. In the former the detected light is incoherent and weak whereas in the latter the coherence properties of the illumination source, typically a laser, dictate the coherence of detected light, but both are dependent on the quality of the illuminating light sheet. Image formation in both schemes can be understood as the convolution of the light sheet with the specimen distribution. In this paper we explore wavefront shaping for the enhancement of light sheet microscopy with scattered light. We show experimental verification of this result, demonstrating the use of the propagation invariant Bessel beam to extend the field of view of a high resolution scattered light, light sheet microscope and its application to imaging of biological super-cellular structures with sub-cellular resolution. Additionally, complementary scattering and fluorescence imaging is used to characterize the enhancement, and to develop a deeper understanding of the differences of image formation between contrast mechanisms in light sheet microscopy.

Citation Download Citation

J. Nylk, C. Mitchell, T. Vettenburg, F. J. Gunn-Moore, and K. Dholakia "Wavefront shaping of a Bessel light field enhances light sheet microscopy with scattered light", Proc. SPIE 8949, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXI, 89490V (12 March 2014); https://doi.org/10.1117/12.2038525

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available