Three-dimensional localization microscopy by incoherent holography

Abhijit Marar; Peter Kner

doi:10.1117/12.2507060

22 February 2019 Three-dimensional localization microscopy by incoherent holography

Abhijit Marar, Peter Kner

Proceedings Volume 10884, Single Molecule Spectroscopy and Superresolution Imaging XII; 108840I (2019) https://doi.org/10.1117/12.2507060
Event: SPIE BiOS, 2019, San Francisco, California, United States

Abstract

Super-resolution imaging of volumes as large as whole cells in three-dimensions (3D) is required to reveal unknown features of cellular organization which cannot be resolved by conventional fluorescence microscopy. We propose a new 3D high resolution imaging technique based on the principles of single-molecule localization microscopy (SMLM) and fluorescence incoherent correlation holography (FINCH). FINCH enables hologram acquisition and three-dimensional (3D) imaging of large objects emitting incoherent light. This technique combines FINCH and SMLM to enable single-molecule volumetric imaging over large axial ranges without scanning the sample using a simple and robust setup, hence making it a viable solution for whole cell super-resolution imaging of biological samples. Here, we present the underlying theory and simulations demonstrating the extended depth of field. We image a single 0.2-μm fluorescent microsphere using this approach and discuss the signal-to-noise ratio (SNR) requirements for an experimental implementation.

Citation Download Citation

Abhijit Marar and Peter Kner "Three-dimensional localization microscopy by incoherent holography", Proc. SPIE 10884, Single Molecule Spectroscopy and Superresolution Imaging XII, 108840I (22 February 2019); https://doi.org/10.1117/12.2507060

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