Applications in Confocal Microscopy
Fluorescent Protein Literature Sources
Dramatically expanding the ability to visualize molecular events with high temporal and spatial resolution in the optical microscope, fluorescent proteins represent perhaps the most important probes ever developed for investigations in cellular and plant biology. With the emergence of the Internet as a practical vehicle for the international dissemination of information, a majority of the scientific publications have begun to make their content available on-line in the form of abstracts, HTML documents, and downloadable portable document format (PDF) files that can be viewed and printed locally. The references listed in the following sections contain links to indexing services in order to provide investigators with a quick pathway to the original literature targeting fluorescent proteins.
General Fluorescent Protein References - The disciplines of cellular and molecular biology are being rapidly and dramatically transformed by the application of fluorescent proteins developed from marine organisms as fusion tags to track protein behavior in living cells. The most widely used of these probes, green fluorescent protein, can be attached to virtually any target of interest and still fold into a viable fluorescent species. The resulting chimera can be employed to localize previously uncharacterized proteins or to visualize and track known proteins to further understand critical events at the cellular and molecular levels. This section features a bibliography of literature sources for review articles and original research reports on the discovery, applications, and continued development of fluorescent proteins.
Biosensor Fluorescent Protein References - By directly coupling the advanced microscopic technique of fluorescence resonance energy transfer (FRET) to the ubiquitous green fluorescent protein and its multispectral variants fused with selected biopolymers, ingenious researchers have created a new class of biosensors capable of elucidating mechanisms of signaling, enzymatic cleavage, and other critical functions in living cells. This section features a bibliography of literature sources for review articles and original research reports on the construction, applications, and continued development of biosensor fluorescent proteins.
Optical Highlighter Fluorescent Protein References - The ability to selectively initiate or alter fluorescence emission profiles in photoconversion optical highlighter proteins renders these probes as excellent tools for exploring protein behavior in living cells. As the fluorescence intensity (or color spectrum) of highlighters occurs only after photon-mediated conversion, newly synthesized non-photoactivated protein pools remain unobserved and do not complicate experimental results. This section lists sources for review articles and original research reports on optical highlighter fluorescent proteins.
Fluorescent Protein Photobleaching References - The field of cell biology is rapidly being transformed by the application of fluorescent proteins as fusion tags to track dynamic behavior in living cells. In this regard, fluorescence recovery after photobleaching (FRAP) is often employed to selectively destroy fluorescent molecules within a region of interest with a high-intensity laser, followed by monitoring the recovery of new fluorescent molecules into the bleached area over a period of time with low-intensity laser light. The resulting information can be used to determine kinetic properties, including the diffusion coefficient, mobile fraction, and transport rate of the fluorescently labeled molecules. The selected references in this section point to important literature sources for information on FRAP with fluorescent proteins.
Fluorescent Protein FRET References - Understanding the dynamic interactions between proteins within living cells is fundamental to a basic knowledge of the underlying concepts that guide molecular and cellular biology. Over the past few years, the rapid development of fluorescent proteins and their application as fusion products and biosensors has significantly expanded the molecular toolkit available for probing the mysteries of cellular physiology and pathology. In this regard, fluorescence (or Förster) resonance energy transfer (FRET) is emerging as a powerful optical microscopy technique for examining physiological processes with high temporal and spatial resolution. The references listed in this section highlight important literature sources for review articles and original research reports on the construction and applications of fluorescent proteins for resonance energy transfer experiments.
Fluorescence Correlation Spectroscopy - Fluorescence correlation spectroscopy (FCS) is a powerful technique used to examine rapid fluctuations in fluorescence emission induced by low concentrations of diffusing labeled proteins and smaller molecules within a restricted volume. The dual-color analog, fluorescence cross-correlation spectroscopy (FCCS) is useful for determining molecular interactions and colocalization between two species emitting at different wavelengths. This section features a bibliography of literature sources for review articles and original research reports focused on fluorescence correlation (and cross-correlation) spectroscopy.