CCD Cameras for Fluorescence Imaging of Living Cells

Abstract

INTRODUCTIONCharged-coupled device (CCD) cameras are commonly used to acquire wide-field, spinning-disk confocal, and total internal reflection fluorescence (TIRF) microscopy images. Collecting digital images for a live cell fluorescence microscopy experiment requires balancing the desire to acquire sufficiently high-quality images with the need to minimize exposure of the specimen to light to reduce phototoxicity and photobleaching. The choice of CCD camera and the camera acquisition settings are critical determinants in the quality of a digital image that one can acquire from a given fluorescent specimen. In this article, we discuss the characteristics of CCD cameras, how they affect digital-image quality, and how to properly use a CCD camera for fluorescence live cell imaging experiments.