Two photon fluorescence background rejection by differential aberration imaging

Aymeric Leray; Kyle Lillis; Jerome Mertz

doi:10.1117/12.707443

9 February 2007 Two-photon fluorescence background rejection by differential aberration imaging

Aymeric Leray, Kyle Lillis, Jerome Mertz

Proceedings Volume 6467, MEMS Adaptive Optics; 646704 (2007) https://doi.org/10.1117/12.707443
Event: MOEMS-MEMS 2007 Micro and Nanofabrication, 2007, San Jose, California, United States

Abstract

We present a simple and robust way to reject out-of-focus background when performing deep two-photon excited fluorescence (TPEF) imaging in thick tissue. The technique is based on the use of a deformable mirror (DM) to introduce illumination aberrations that preferentially degrade TPEF signal while leaving TPEF background relatively unchanged. A subtraction of aberrated from unaberrated images leads to background rejection. We present a heuristic description of our technique, which we corroborate with experiment. Images of a labeled mouse olfactory bulb are compared with standard TPEF microscopy images, demonstrating significant out of focus TPEF background rejection with our technique. Finally we improve our technique by developing a faster aberration modulation mechanism that performs background subtraction line by line rather than frame by frame. In this manner, the overall image acquisition rate of our technique is the same as that of a standard TPEF microscope.

Citation Download Citation

Aymeric Leray, Kyle Lillis, and Jerome Mertz "Two-photon fluorescence background rejection by differential aberration imaging", Proc. SPIE 6467, MEMS Adaptive Optics, 646704 (9 February 2007); https://doi.org/10.1117/12.707443

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

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
10 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Luminescence

Microscopes

Deformable mirrors

Laser scattering

Calcium

Objectives

Tissues

Show All Keywords

Keywords/Phrases

Search In:

Publication Years