Encoded multisite two-photon microscopy.
Proceedings of the National Academy of Sciences. 2013-06-24; 110(32): 13138-13143
DOI: 10.1073/pnas.1307818110
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1. Proc Natl Acad Sci U S A. 2013 Aug 6;110(32):13138-43. doi:
10.1073/pnas.1307818110. Epub 2013 Jun 24.
Encoded multisite two-photon microscopy.
Ducros M(1), Goulam Houssen Y, Bradley J, de Sars V, Charpak S.
Author information:
(1)Institut National de la Santé et de la Recherche Médicale U603, Paris 75006,
France.
Comment in
Microsc Res Tech. 2013 Oct;76(10):985-7.
The advent of scanning two-photon microscopy (2PM) has created a fertile new
avenue for noninvasive investigation of brain activity in depth. One principal
weakness of this method, however, lies with the limit of scanning speed, which
makes optical interrogation of action potential-like activity in a neuronal
network problematic. Encoded multisite two-photon microscopy (eMS2PM), a scanless
method that allows simultaneous imaging of multiple targets in depth with high
temporal resolution, addresses this drawback. eMS2PM uses a liquid crystal
spatial light modulator to split a high-power femto-laser beam into multiple
subbeams. To distinguish them, a digital micromirror device encodes each subbeam
with a specific binary amplitude modulation sequence. Fluorescence signals from
all independently targeted sites are then collected simultaneously onto a single
photodetector and site-specifically decoded. We demonstrate that eMS2PM can be
used to image spike-like voltage transients in cultured cells and fluorescence
transients (calcium signals in neurons and red blood cells in capillaries from
the cortex) in depth in vivo. These results establish eMS2PM as a unique method
for simultaneous acquisition of neuronal network activity.
DOI: 10.1073/pnas.1307818110
PMCID: PMC3740851
PMID: 23798397 [Indexed for MEDLINE]