Single molecule fluorescence saturation spectroscopy - Archive ouverte HAL Access content directly
Conference Papers Year :

Single molecule fluorescence saturation spectroscopy

(1) , (1)
1

Abstract

Saturation spectroscopy is a powerful method to investigate photophysical parameters of single fluorescent molecules. Nevertheless, the impact of a gradual increase, over a broad range, of the laser excitation on the intramolecular dynamics is not completely understood, particularly concerning their fluorescence emission (the so-called brightness). As we have presented in a previous paper [1], we interpret the evolution of the brightness with the laser power by cascade absorption of two and three photons within a five-level molecular system. This multi-photon consecutive absorption leads us to reconsider the common expression of the saturation curve of fluorescent molecule. Furthermore, this multi-photon absorption process also affects the observation volume of microscope. So, in this paper we propose to interpret the size increase of the confocal observation volume according to simulations based upon two often used expressions of the Point Spread Functions (PSF) in fluorescence microscopy.
Not file

Dates and versions

hal-02531216 , version 1 (03-04-2020)

Identifiers

Cite

Rodolphe Jaffiol, Pascale Winckler. Single molecule fluorescence saturation spectroscopy. SPIE BiOS, Feb 2014, San Francisco, United States. pp.89500C, ⟨10.1117/12.2037029⟩. ⟨hal-02531216⟩

Collections

CNRS UTT
19 View
0 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More