Confocal Microscopy

Confocal Microscopy

Confocal Microscopy

The optical imaging technique of Laser Confocal Scanning Microscopy (LCSM) is used to increase the optical resolution and contrast of an image. Although laser scanning is most commonly used, the scanning process is relatively slow and requires high light levels that may produce photo-damage, particularly in living specimens. Spinning disk confocal microscopy uses a wide illumination field that is projected through microscopic apertures in a rotating disk to generate a scan pattern. This parallelizes the scanning process, making it faster. Confocal imaging is compatible with many spectral applications including multi-fluorescence imaging, time-lapse imaging, FLIM, FRAP, and FCS measurements.

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Confocal Microscopy

Basics in Microscopy

Basics in Microscopy

Knowledge in optics and contrasting methods is essential in understanding the fundamentals of microscopic imaging. Physical principals surrounding the operation of a microscope include resolution, numerical aperture, depth of field, image brightness, objective working distance, field of view, conjugate planes, and the useful magnification range. Contrasting methods like phase contrastmodulation contrastdifferential interference contrast, often aid scientist in visualizing living samples as these methods convert phase shifts into intensity. Staining and fluorescence techniques, like immunofluorescence or the use of fluorescent proteins, are used to make selected structures or proteins visible.

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Confocal Microscopy

Neuroscience

Neuroscience

Neuroscience is the multidisciplinary branch of biology in which the fundamental properties of neurons, the organization of neural networks, and their behavioral and sensory functional products are studied. Because the functions of neural networks are inextricably linked to their spatial organization, neuroscience research is heavily dependent on a variety of imaging techniques. For cellular and tissue level investigations, fluorescence microscopy is the most instructive and widely applicable of these techniques. Optogenetic techniques provides researchers with ways of externally modulating neuronal network function using light as an intermediary.

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Confocal Microscopy

Intravital Imaging

Intravital Imaging

Intravital imaging is a method that allows for visualization of biological processes in live animals (in vivo) with sufficient resolution to pick out cellular behaviors and molecular signals under lying them. This method of microscopy can be performed using several light microscopy techniques including wide-field fluorescence, confocal, multi photon, and spinning disk microscopy. What technique is implemented for intravital imaging is based on what penetration of depth is needed to image an area and the amount cell-cell interaction details required.

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Confocal Microscopy

Cell Biology

Cell Biology

Cell biology is a field where cell structure and function are studied. Because function depends on spatial organization microscopic imaging is required to understand it. Cell biologist concern themselves with understanding physiological properties, metabolic processes, signaling pathways, chemical composition, and interactions of the cells with their environment.Cell Biology is closely related to other field such as immunology, genetics, biochemistry, and molecular biology.

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Confocal Microscopy

Quantitative Fluorescence

Quantitative Fluorescence

Quantitative fluorescence microscopy techniques offer a researcher powerful tools to understanding physiological and pathological roles of molecular processes in living cells. For example, to understand regulation of protein expression, it is necessary to know the amounts of protein expressed as a function of time. However the fluorescence levels in a field of view depend not only on the amount of protein present, but also on external factors, including the excitation light intensity. Techniques that are based in fluorescence such as FLIM, FCS, FCCS, FRET, and FRAP aid in the measurement and quantification of data.

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