Confocal microscopy and its application on forensic science Essay Example
Соnfосаl Miсrоsсорy and its Аррliсаtiоn on Fоrеnsiс Sсiеnсе 5
Соnfосаl Miсrоsсорy and its Аррliсаtiоn on Fоrеnsiс Sсiеnсе
According to Price and Jerome (2011), confocal microscopy, also referred as Confocal Laser Scanning Microscopy (CLSM), is the technique of optimal imaging in order to increase the optimal contrast and resolution of a micrograph through the means of addition of a spatial pinhole, which is placed at the confocal plane of the lens to facilitate the elimination of focus light. Notably, the over the past few years, confocal microscopy has gained popularity especially in the fields of industrial and scientific communities with typical applications in semiconductor inspection, materials science, and life sciences (Stevens et al. 2015). In this way, confocal microscopy helps in enabling the reconstruction of structure with three dimensions from the images that have been obtained through collection of sets of images in the optical sectioning process at different depths within a thick object. Critical to note is that the conventional microscopes see into a specimen as far as the penetration of the light. However, the confocal microscopes provide images one level at a given time. In this way, the application and the use of CLSM help in achieving a highly limited and controlled depth of focus.
Evidently, due to advanced technology, the current instruments in the field of microscopy are highly evolved since the introduction of earlier versions (Stevens et al. 2015). However, the principle of imaging through confocal microscopy, which was started and patented in 1957 by Marvin Minsky, has been employed in many modern confocal microscopes.
In any conventional widefield microscope, the entire specimens are bathed in light that are sourced from xenon or mercury (Price & Jerome 2011). The images are then directly viewed by the eye or are projected onto photographic films or image capture devices (Stevens et al. 2015). Unlike the conventional microscopes, the method of imaging formations through confocal microscopes is fundamentally different. In this new form of microscopy, illumination is achieved by caning more or even one focused beams of light, mostly from across the specimen, arc-discharge source or from a laser. The point that is illuminated is brought to focus in the specimen though the objective lens as well as laterally scanning using forms of scanning devices that are controlled by a computer. Further, the set and sequences of points from the light, sourced from the specimen are detected though the photomultiplier tube (PMT) by the help of a pinhole or a slit (Stevens et al. 2015). In this way, from the PMT, the output is built into the image before being displayed by a computer. Although one can view unstained specimens with light that is reflected back from a specimen, the unstained specimens are usually labeled with either one or more fluorescent probes.
Application of Confocal Microscopy in Forensic Science
Confocal microscopy has been one of the key components that have revolutionized forensic science especially in the recent years (Pawley 2016). Notably, as an example, the biological laser scanning confocal microscopy is dependent on the fluorescence as the mode of imaging due to the level of sensitivity that that the technique is able to afford as well as the specific target of structural components including the dynamic processes in living and chemically fixed tissues and cells (Pawley 2016). Many fluorescent probes are built around the synthetic aromatic organic chemicals that are designed in order to bind with a biological macromolecule such as nucleic acid or a protein as well as localizing within a particular structural region, such as the mitochondria, Golgi apparatus, cytoskeleton, endoplasmic reticulum, and nucleus (Stevens et al. 2015). Confocal microscopy is also applied in monitoring localized environmental variables and dynamic processes such as the concentrations of reactive oxygen species, inorganic metallic ions, the pH, and membrane potential, among others.
. Boston, MA, Springer US.Handbook of Biological Confocal MicroscopyPawley, J. B. (2016).
Price, R. L., & Jerome, W. G. (. (2011). Basic confocal microscopy. New York, NY, Springer.
Stevens, J. K., Mills, L. R., & Trogadis, J. E. (2015). Three-Dimensional Confocal Microscopy:
Volume Investigation of Biological Specimens. Burlington, Elsevier Science.
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