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The Application of Fluorescent Quantum Dots to Confocal, Multiphoton, and Electron Microscopic Imaging

From the Center for Research on Biological Systems and the National Center for Microscopy and Imaging Research, University of California, San Diego, USA

Correspondence: Address correspondence to: Thomas J. Deerinck, National Center of Microscopy and Imaging Research (NCMIR), Center for Research on Biological Systems, University of California, San Diego (UCSD) School of Medicine, 1000 BSB, 9500 Gilman Drive, MC0608, La Jolla, CA 92093-0608; e-mail: deerinck{at}ncmir.ucsd.edu.

Fluorescent quantum dots are emerging as an important tool for imaging cells and tissues, and their unique optical and physical properties have captured the attention of the research community. The most common types of commercially available quantum dots consist of a nanocrystalline semiconductor core composed of cadmium selenide with a zinc sulfide capping layer and an outer polymer layer to facilitate conjugation to targeting biomolecules such as immunoglobulins. They exhibit high fluorescent quantum yields and have large absorption cross-sections, possess excellent photostability, and can be synthesized so that their narrow-band fluorescence emission can occur in a wide spectrum of colors. These properties make them excellent candidates for serving as multiplexing molecular beacons using a variety of imaging modalities including highly correlated microscopies. Whereas much attention has been focused on quantum-dot applications for live-cell imaging, we have sought to characterize and exploit their utility for enabling simultaneous multiprotein immunolabeling in fixed cells and tissues. Considerations for their application to immunolabeling for correlated light and electron microscopic analysis are discussed.

Key Words: Quantum dot • light microscopy • fluorescence microscopy • electron microscopy • immunolabeling • multiphoton microscopy

Abbreviations: EM, electron microscopy • HeLa, Henrietta Lacksm • IgG, immunoglobulin G • LR, London Resin • NA, numerical aperture • QD, quantum dot • RFL6, rat fetal lung fibroblasts • TIRF, total internal reflection fluorescence microscopy • UV, ultraviolet

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