Fluorescence lifetime correlation spectroscopy for precise concentration detection in vivo by background subtraction

Maria Gärtner; Jörg Mütze; Thomas Ohrt; Petra Schwille

doi:10.1117/12.831572

7 July 2009 Fluorescence lifetime correlation spectroscopy for precise concentration detection in vivo by background subtraction

Maria Gärtner, Jörg Mütze, Thomas Ohrt, Petra Schwille

Author Affiliations +

Proceedings Volume 7368, Clinical and Biomedical Spectroscopy; 73681V (2009) https://doi.org/10.1117/12.831572
Event: European Conferences on Biomedical Optics, 2009, Munich, Germany

Abstract

In vivo studies of single molecule dynamics by means of Fluorescence correlation spectroscopy can suffer from high background. Fluorescence lifetime correlation spectroscopy provides a tool to distinguish between signal and unwanted contributions via lifetime separation. By studying the motion of the RNA-induced silencing complex (RISC) within two compartments of a human cell, the nucleus and the cytoplasm, we observed clear differences in concentration as well as mobility of the protein complex between those two locations. Especially in the nucleus, where the fluorescence signal is very weak, a correction for background is crucial to provide reliable results of the particle number. Utilizing the fluorescent lifetime of the different contributions, we show that it is possible to distinguish between the fluorescent signal and the autofluorescent background in vivo in a single measurement.

Citation Download Citation

Maria Gärtner, Jörg Mütze, Thomas Ohrt, and Petra Schwille "Fluorescence lifetime correlation spectroscopy for precise concentration detection in vivo by background subtraction", Proc. SPIE 7368, Clinical and Biomedical Spectroscopy, 73681V (7 July 2009); https://doi.org/10.1117/12.831572

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