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1 May 2006 High-speed addressable confocal microscopy for functional imaging of cellular activity
Vivek Bansal, Saumil Patel, Peter Saggau
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Journal of Biomedical Optics, Vol. 11, Issue 3, 034003 (May 2006). https://doi.org/10.1117/1.2209562
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.
©(2006) Society of Photo-Optical Instrumentation Engineers (SPIE)
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Vivek Bansal, Saumil Patel, and Peter Saggau "High-speed addressable confocal microscopy for functional imaging of cellular activity," Journal of Biomedical Optics 11(3), 034003 (1 May 2006). https://doi.org/10.1117/1.2209562
Published: 1 May 2006
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KEYWORDS
Confocal microscopy
Imaging systems
Mirrors
Neurons
Luminescence
Digital micromirror devices
Signal to noise ratio
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