Basics in 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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Basics in 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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Basics in 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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Basics in Microscopy

Digital Microscopy

Digital Microscopy

Digital Microscopy is a combination of the advances that have been made in optics and digital cameras to output an image on a monitor. Typical microscope set ups for digital microscopy do not include eye pieces to view the sample directly like what is traditionally done in stereo microscopy.

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Basics in Microscopy

Stereo Microscopy

Stereo Microscopy

Stereo microscopy is an important tool in understanding and inspecting microscopic structures for their spatial extent and nature as well as offering great advantage of providing a 3D view of the sample. There are a number of applications where stereomicroscopes are indispensable such as industrial production, quality assurance, materials research, forensics, biotechnology, genetics, and biomedical research. The stereo microscope is typically used to study the surfaces of solid specimens and to provide visual guidance for dissection, microsurgery, circuit board manufacture and fracture inspection procedures.

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