Work

Second-Harmonic Generation Imaging of Membrane Potential with Photon Counting

Year: 2008

Type: article

Abstract: Second-harmonic generation (SHG) can be used for imaging membrane potential in neurons, but poor signal-to-noise (S/N) limits accurate measurements of small voltage transients. We use photon counting to improve the S/N of weak SHG signal detection. Photon counting generates shot-noise limited and integrable signals, eliminates pulse-to-pulse variation, and built-in discriminators reduces the background to practically zero. In single trials, by using photon counting, we obtain a more than a twofold S/N increase over analog voltage detection. Trial-to-trial variability is also reduced by 50%. Finally, we show that, using photon counting, the kinetics of fast events such as action potentials can be recorded more accurately.

Source: Microscopy and Microanalysis

DOI: 10.1017/s1431927608080811

Authors: Jiang Jiang, Rafael Yuste

Institutions: Howard Hughes Medical Institute, Columbia University

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: Gatsby Charitable Foundation, National Eye Institute

Award: Imaging Functional Connectivity in Visual Cortex