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In general, the maximum magnification of a confocal microscope is 1000x, assuming the use of a combination of a 100x objective and a 10x ocular. It is dependent on the limitations of the mounting type used, thickness of tissue, and optics of the system. Let’s discuss this in detail.
Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser scanning confocal microscopy (LSCM), is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a spatial pinhole to block out-of-focus light in image formation. [ 1 ]
A confocal microscope is a type of microscope that uses laser light to produce high-resolution images of biological samples. It works by illuminating the sample with a laser, and using a series of lenses and mirrors to focus the light onto a specific plane within the sample.
Confocal microscopy offers several advantages over conventional optical microscopy, including shallow depth of field, elimination of out-of-focus glare, and the ability to collect serial optical sections from thick specimens.
If a pinhole is placed here, blurred light generated outside the focal plane that cannot be focused is blocked by the pinhole, allowing light from a single point on the focal plane to be selectively detected. This is the principle of confocal microscopy.
In vivo confocal microscopy (IVCM) is an emerging noninvasive imaging and diagnostic tool, which enables morphological and quantitative analysis of ocular surface microstructure. The principal of confocal microscopy was patented in 1957 by Marvin Minsky .
• The importance of understanding the microscope objective –Magnification –Resolution –Abberations –Working distance • Confocal Microscopy • Two-photon Microscopy • Spinning disc confocal • Summary
In practice, the best horizontal resolution of a confocal microscope is about 0.2 microns, and the best vertical resolution is about 0.5 microns. I wrote a brief discussion here of the difference between resolution and magnification. How big is a confocal microscope?
Critical Aspects of Confocal Microscopy. Laser alignment, photobleaching, probes, magnification, and other important concepts. Three-Color Confocal Imaging. Confocal microscopy can be used to produce images of triple-labeled fluorescent samples. Resonant Scanning in Laser Confocal Microscopy
The complete generation of the two-dimensional object information from the focal plane (object plane) of a confo-cal LSM essentially comprises three process steps: Line-by-line scanning of the specimen with a focused laser beam deflected in the X and Y directions by means of two galvanometric scanners.