MicroscopicImagingFluorescence-I

MicroscopicImagingFluorescence-I - Modern Optical...

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Andres Kriete Bioengineering, Science and Health Systems Drexel University, Philadelphia, PA Modern Optical Microscopy: Fluorescence and Image Fidelity
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Introduction into microscopy Fluorescence imaging & markers Imaging Devices Systems Theory
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TEM vs. Light Microscope What you can see with a light microscope is limited by the wavelength of light. TEMs use electrons as "light source" and their much lower wavelength makes it possible to get a resolution a thousand times better than with a light microscope. Light Microscop e TEM
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Basic Concepts Transmission electron microscopy (TEM) is a microscopy technique whereby a beam of electrons is transmitted through an ultra thin specimen, interacting with the specimen as it passes through. http://www.unl.edu/CMRAcfem/temoptic.htm
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Images by TEM Mitochondrium ER Virus Nucleus with chromatin
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Dimensions http://learn.genetics.utah.edu/content/begin/cells/scale/
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How does it work? A "light source" at the top of the microscope emits the electrons that travel through vacuum in the column of the microscope. (electron gun) The TEM uses electromagnetic lenses to focus the electrons into a very thin beam by use of the condenser lens.
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Working areas of imaging systems
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Abbe’s Law Ernst Abbe together with Carl Zeiss published a paper in 1877 defining the physical laws that determined resolving distance of an objective. Known as Abbe’s Law. Minimum resolved distance (d) is related to wavelength λ devided by numerical aperture with angle of light cone (theta).
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Diffraction-limited Resolution
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Microscope Setup Dichroic Objective Arc Lamp Emission Filter Excitation Diaphragm Ocular Excitation Filter EPI-Illumination
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This note was uploaded on 07/08/2011 for the course BM 501 taught by Professor Kop during the Spring '11 term at Bloomsburg.

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MicroscopicImagingFluorescence-I - Modern Optical...

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