Sunday, November 26, 2006

Confocal Microscopy Internet Resources

General Information - The growing utility and popularity of confocal microscopy has resulted in a dramatic increase in applications during recent years and, correspondingly, so have the number of sites on the Internet that address the subject. The links below constitute a selected collection from the best websites available today that provide general information about the history, theory, and practice of this exciting technique. Microscopists at all levels, from beginner to professional, may benefit from visiting many of these resources.

Antibodies - Antibodies are a large family of glycoproteins produced by specialized cells of the vertebrate immune system in response to foreign molecules (antigens). The interaction of an antibody with an antigen is the foundation upon which all immunochemical techniques have been developed. In confocal microscopy, such techniques are often extremely useful, enabling the researcher to localize antigens in cell cultures and tissue sections by utilizing antibodies labeled with fluorescent dyes that can be readily visualized with a confocal microscope. Also, in order to obtain a stronger signal, oftentimes an unlabeled primary antibody is used in conjunction with a labeled secondary antibody that binds to it. The resources provided in this section include many leading and emerging producers and distributors of primary and secondary antibodies.

Area Array (CCD and CMOS) Detectors - In recent years, optical microscopy has slowly migrated from a dependence on traditional photomicrography using emulsion-based film and has become increasingly reliant on technology that produces electronic images. Indeed, the choice of an imaging device is a critical decision for modern microscopists, but the range of light detection methods and the tremendous variety of imaging devices available can make the selection process difficult. This collection of area array detector resources is designed to simplify this process, providing links to many of the best sites on the Internet that offer CCD and CMOS detectors, as well as other imaging solutions to microscopists.

Digital Image Processing and Analysis - Despite the availability of the latest state of the art hardware, microscopists are often required to utilize advanced software for acquisition, processing, archiving, and retrieval of digital images in order to examine and reveal certain fine specimen details that would otherwise remain unseen. A wide range of companies offer such specialized software, providing consumers with a host of options for meeting the specific digital image processing and analysis requirements created by their application needs.

Fluorescence Filters - Fluorescence microscopy relies heavily on the ability to select a specific wavelength region for excitation of the specimen and gathering secondary emission during image formation. Recent advances in interference filter design have resulted in highly accurate filters that cover a wide range of bandpass profiles, ranging from just a few to tens and hundreds of nanometers. Listed below is a compilation of manufacturers that design, manufacture, and supply fluorescence filters to the microscopy community. The products that they offer will meet most system requirements, but if a specialized application necessitates a filter with an unusual spectral range or other unique specifications, many of the companies will fabricate custom filters.

Fluorescent Probes - Fluorescence is the property exhibited by many molecules of absorbing light at a particular wavelength and subsequently emitting light of longer wavelength after a brief interval in time. Some atoms and molecules fluoresce naturally, but others must be artificially altered or marked in some way to instigate the phenomenon. In confocal microscopy, fluorescent probes play a paramount role in the detection of anatomical structure and physiological reactions occurring in living cells. An extensive array of fluorescent probes are available from a number of distributors, the best of which have been compiled into the following list of Internet resources.

Fluorescent Protein Educational Websites - The discovery and development of fluorescent proteins from jellyfish and other marine organisms has drastically transformed cell research in recent years, providing life scientists with a minimally invasive means of studying protein dynamics and function in live cells and tissues. The websites listed in this section are an excellent educational starting point for individuals interested in broadening their knowledge of these unique investigative tools. Within the featured resources, information related to many different aspects of fluorescent proteins is available, including their history, attributes, and applications.

Fluorescent Protein Principle Investigators - Many of the scientists involved with research targeting various aspects of cell biology are using fluorescent proteins as imaging probes for cell structure, function, and dynamics. Several of the principle investigators have built extensive websites detailing their laboratories, and these sites are quite useful to visitors interested in learning more about this exciting and rapidly evolving research arena. Included in the information on a majority of the websites linked below are the current research interests, curriculum vitas, publications, lists of laboratory personnel, contact information, educational tutorials, image galleries, and digital videos.

Fluorescent Protein Vector Commercial Sources - A variety of fluorescent proteins, available as recombinant DNA plasmid vectors designed for transfection of mammalian cells or transformation of bacteria, are commercially available from a number of distributors. Most of the vectors containing fluorescent protein DNA sequences have been codon-optimized for expression in mammalian cells and contain antibiotic genes for selection of stable mutants having relatively constant expression levels. The vectors often contain multiple cloning sequences that enable researchers to easily insert their gene of interest for fusion to the fluorescent protein. Other common features in fluorescent protein vectors include a human cytomegalovirus (CMV) promoter, a Kozak translation initiation site, an early mRNA polyadenylation signal, and a bacterial antibiotic gene.

FluoViewTM Users Internet Resources - The FluoViewTM laser scanning confocal microscope is a key piece of equipment in laboratories around the world, being utilized for applications that range from quantitative cellular analysis to neuroanatomy, toxicology, molecular genetics, and zoological studies. A sampling of the many renowned universities and private research institutions that have benefited from this exciting Olympus technology is provided through the Internet links in this section.

Laser Systems - At one time limited only to the subject matter of science fiction stories, since their invention in the early 1960s, society has become increasingly dependent on lasers, utilizing the tools in a wide variety of instrumentation that ranges from barcode scanners and compact disk players to surgical equipment and interferometers. Indeed, lasers are the most common light source employed for scanning confocal fluorescence microscopy and, therefore, microscopists utilizing this technique should have a solid understanding of these powerful, radiation-emitting devices.

Live-Cell Imaging - One of the foremost targets in the life sciences is to understand the structure, function, and behavior of living organisms, and with evolving advances in technology, such as the development of confocal microscopy and fluorescent probes, it has become possible to pursue this goal at the cellular and subcellular levels. Still, working with and imaging live cells can be a complex, if not daunting, task to microscopists unfamiliar with the techniques and tools that are available. The following is a compilation of resources that offer overviews, background information, interactive forums, frequently asked questions, protocols, and hints that should aid any microscopist attempting to enter into this important, burgeoning field.

Live-Cell Imaging Specimen Chambers - The demands of modern confocal microscopy, especially those experiments involving the imaging of living cells and tissues, require that researchers take special precautions with their specimens. Indeed, simple microscope slides are unsuitable for many applications, resulting in the development of a broad range of specimen chambers, which can often supply the neccesary flexibility for live-cell imaging. The list of resources in this section exemplifies the great variety of specimen chambers that are commercially available, and is designed to help visitors locate the products that are best suited for their specific scientific pursuits.

Microscopy Courses and Workshops - A number of universities, research institutions, and organizations offer excellent courses, workshops, conferences, and symposia relating to confocal microscopy and its applications. The compilation of sites provided in this section are those belonging to groups that present these offerings on a regular basis, such as annually or biannually. Other highly useful courses and instructional events occur on a more sporadic schedule, and, therefore, this list of resources should be considered as a starting, rather than an ending, point for those seeking educational opportunities in the field.

Photometric Detectors - Photometric detectors, such as photomultiplier tubes and avalanche photodiodes, are the preferred photon detectors for a number of applications, including laser scanning confocal microscopy. A large number of companies, therefore, offer these forms of technology to the research and teaching community. This list of resources highlights the major developers, manufacturers, and suppliers of point source detectors, as well as some of the best sites on the Internet that offer background and conceptual information relating to them.

Three-Dimensional Volume Rendering - In order to obtain the most significant amount of information from the two-dimensional image stacks acquired through confocal microscopy, three-dimensional volume rendering software is necessary. In recent years, a vast array of software programs have been developed to help microscopists meet this critical need, many of which are open-source and freely available. Featured within this selection of three-dimensional volume rendering resources are websites that describe and distribute some of the most powerful software tools available to microscopists.

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