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Journal of Biomedical Optics

High-speed addressable confocal microscopy for functional imaging of cellular activity
Author(s): Vivek Bansal; Saumil Patel; Peter Saggau
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Paper Abstract

Due to cellular complexity, studying fast signaling in neurons is often limited by: 1. the number of sites that can be simultaneously probed with conventional tools, such as patch pipettes, and 2. the recording speed of imaging tools, such as confocal or multiphoton microscopy. To overcome these spatiotemporal limitations, we develop an addressable confocal microscope that permits concurrent optical recordings from multiple user-selected sites of interest at high frame rates. Our system utilizes acousto-optic deflectors (AODs) for rapid positioning of a focused laser beam and a digital micromirror device (DMD) for addressable spatial filtering to achieve confocality. A registration algorithm synchronizes the AODs and DMD such that point illumination and point detection are always colocalized in conjugate image planes. The current system has an adjustable spatial resolution of ~0.5 to 1 µm. Furthermore, we show that recordings can be made at an aggregate frame rate of ~40 kHz. The system is capable of optical sectioning; this property is used to create 3-D reconstructions of fluorescently labeled test specimens and visualize neurons in brain slices. Additionally, we use the system to record intracellular calcium transients at several sites in hippocampal neurons using the fluorescent calcium indicator Oregon Green BAPTA-1.

Paper Details

Date Published: 1 May 2006
PDF: 9 pages
J. Biomed. Opt. 11(3) 034003 doi: 10.1117/1.2209562
Published in: Journal of Biomedical Optics Volume 11, Issue 3
Show Author Affiliations
Vivek Bansal, Baylor College of Medicine (United States)
Saumil Patel, Baylor College of Medicine (United States)
Peter Saggau, Baylor College of Medicine (United States)

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