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Proceedings Paper

Scanning algorithms in high-sensitivity two-photon excitation microscopy for single-molecule investigations
Author(s): Giuseppe Chirico; Gabriele Malengo; Roberto Milani; Fabio Cannone; Silke Krol; Alberto Diaspro
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Paper Abstract

We describe and apply two raster scanning algorithms that are suited for fast imaging and for spectroscopy in a two photon microscope. Imaging can be performed at a rate of 1-100 Hz per line with a closed loop piezo-actuator. In order to reach the single molecule sensitivity and to study the dynamics of the fluorescence emission, the detection is performed via avalanche photodiodes. In a slow scanning algorithm we have implemented photon counting histogram and lifetime analysis on the image. In this way we are able to discriminate between local concentration and molecular brightness and to measure lifetimes on extended samples.

Paper Details

Date Published: 21 June 2004
PDF: 8 pages
Proc. SPIE 5323, Multiphoton Microscopy in the Biomedical Sciences IV, (21 June 2004); doi: 10.1117/12.537915
Show Author Affiliations
Giuseppe Chirico, Univ. degli Studi di Milano-Bicocca (Italy)
INFM (Italy)
Gabriele Malengo, Univ. degli Studi di Milano-Bicocca (Italy)
INFM (Italy)
Roberto Milani, Univ. degli Studi di Milano-Bicocca (Italy)
INFM (Italy)
Fabio Cannone, Univ. degli Studi di Milano-Bicocca (Italy)
INFM (Italy)
Silke Krol, Univ. degli Studi di Genova (Italy)
Alberto Diaspro, Univ. degli Studi di Genova (Italy)
Univ. degli Studi di Milano Bicocca (Italy)
INFM (Italy)


Published in SPIE Proceedings Vol. 5323:
Multiphoton Microscopy in the Biomedical Sciences IV
Ammasi Periasamy; Peter T. C. So, Editor(s)

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