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Temporally Gating a Slow-Scan CCD with a Liquid Crystal Shutter

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The application of charge-coupled device (CCD) technology to high-resolution luminescence microscopy is a recent development. The very high sensitivity, linear response, and field uniformity that are characteristic of a slow-scan, scientific CCD camera yield data of sufficient quality to make quantitative image analysis possible. However, the low temporal resolution of current CCD cameras is a major disadvantage. A CCD with a read noise level of less than 10 electrons typically has a readout rate in the 40–200 kHz range, which precludes the acquisition of a time-resolved image integrated over multiple excitation pulses without the use of an external gating device. Imaging in the time domain can potentially increase both the information content and the sensitivity of luminescence microscopy. For example: (1) fluorescent probe distributions can be resolved temporally as well as spatially, yielding improved image contrast; and (2) temporally gated detection of long-lived luminescence discriminates against scattered light and autofluorescence, which can yield improved sensitivity.

Keywords: CCD; Fluorescence microscopy; Liquid crystal shutter; Spectroscopic techniques; Time-resolved luminescence

Document Type: Research Article


Affiliations: Department of Chemistry, University of Arizona, Tucson, Arizona 85721

Publication date: February 1, 1993

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