An evaluation of confocal versus conventional imaging of biological structures by fluorescence light microscopy (original) (raw)

Confocal Microscopy and its Implementation in Different Biological Aspects

2016

Biophysical tools and its techniques are in high demand for the study of molecular mechanisms underlying cell biology, immunology and biomedical research including several diagnostics techniques. Biophysical tools basically include microscopy and imaging hydrodynamics, electrophysiology, modeling and simulation, single molecule techniques and spectroscopy. Among these, microscopy is one of the streams of optics that not only presents a magnified view of cellular structures of bacteria, germs, organs etc. but also ensures micro to nano elaboration of organisms or their parts. In this regard, confocal microscopy has become an essential technique in all the fields of biomedical research including life sciences that produce optical sections of cells and tissues that are totally free from fluorescence. In this review, we described the importance and applications of confocal microscopy along with its implementation in different biological aspects.

Confocal Microscopy of Biological Specimens

Science on Form, 1990

For imaging biological subjects, the confocal bright field method has some advantages over confocal fluorescence: axial resolution is greater, and images can be formed from unstained, living specimens. Techniques for imaging phase information (small changes in refractive index) are also possible and provide yet another realm of useful information about the specimen in addition to its absorption, transmission and reflection properties. Electronic or digital image enhancement techniques can be optimised to produce maximum visibility of detail for a human observer. Finally, optical sections from through-focus series can be combined to produce stereo pairs or stored with depth information and later imaged using a three-dimensional graphics representation.

Advanced microscopy: Laser scanning confocal microscopy

Methods in Molecular Biology, 2011

Fluorescence microscopy is an important and fundamental tool for biomedical research. Optical microscopy is almost non-invasive and allows highly spatially resolved images of organisms, cells, macromolecular complexes, and biomolecules to be obtained. Generally speaking, the architecture of the observed structures is not significantly modified and the environmental conditions can be kept very close to physiological reality. The development of fluorescence microscopy was revolutionized with the invention of laser scanning confocal microscopy (LSCM). With its unique three-dimensional representation and analysis capabilities, this technology gives us a more real view of the world.

Confocal laser scanning microscopy

Bios, Oxford; Springer, New York; Springer, Singapore, 1997

Confocal microscopy is a new science. While the idea for a confocal microscope was first patented by Minsky in 1957, and the first purely analogue mechanical confocal microscope was designed and produced by Eggar and Petran a decade later, it was not until the late seventies, with the advent of affordable computers and lasers, and the development of digital image processing software, that the first single-beam confocal laser scanning microscopes were developed in a number of laboratories and applied to biological and materials specimens. For biologists, a crucial turning point came in 1985 with the publication of six papers from four separate laboratories independently demonstrating the power of the confocal fluorescence microscope to eliminate out-of-focus blur, and thus to obtain three-dimensional (3D) data from intact biological specimens by non-invasive optical sectioning. In a remarkably rapid development, in which both of us were involved, the first commercial confocal laser scanning fluorescence microscopy systems were produced within 2 years of these publications by a small Oxfordshire company which has now become Bio-Rad Microscience Ltd. A number of other companies, including all the major microscope manufacturers worldwide, were swift to produce their own confocal instruments of widely varying designs and capabilities. During the last decade the availability of confocal laser scanning microscopes of ever-increasing power and sophistication has revolutionized the science of microscopy as applied to cell and develop- mental biology, physiology, cytogenetics, diagnostic pathology, and the material sciences. In particular, the ability to obtain a time-series of three-dimensional images from a living specimen, with temporal and spatial resolutions as good as or superior to video microscopy, has opened up new avenues of investigations previously impossible to contemplate. Over the last decade, several excellent books and reviews on confocal microscopy have been published, but there has been a noticeable gap in the availability of a small handbook that introduces the interested student or research worker to this important microscopical technique, and that illustrates how it might benefit their own research. It is this gap that we hope this Handbook, the latest in the Royal Microscopical Society Microscopy Handbooks series, will fill. Starting from first principles, this Handbook explains to the reader what a confocal microscope is, how it is constructed and used what its benefits. are, and. why i.ts imaging performance is superior to'that of a conventional optical mIcroscope. It discusses multiparameter confocal fluorescence microscopy, describes digital image processing and animation of 3D confocal images, illustrates applications of confocal microscopy in both the biomedical and the materials sciences and concludes with a discussion of future developments in this new area of microscopy. Richly illustrated with colour micrographs and diagrams chosen for their clarity and didactic quality, this book also contains an up-to-date bibliography of the most informative publications on confocal microscopy, a catalogue of World Wide Web sites of relevance, and a listing of the names and addresses of confocal microscope and fluorescence filter manufacturers, image processing software vendors, and reagent suppliers. We hope that you will find it useful.