simple microscope objective magnification working distance eyepiece magnification
Post on 21-Dec-2015
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Simple Microscope
• Objective magnification• Working Distance• Eyepiece magnification
http://micro.magnet.fsu.edu/
Illumination (Bright Field)• Simple mirror (historical microscope)• Critical illumination• Koehler Illumination
Summary the field of view should be (reasonably) evenly illuminated the illuminating train should be able to fully illuminate the aperture of an objective of NA = 1.0 the light source should be focused in the object in critical illumination, the light bulb is the light source in Köhler illumination, the light source is an iris diaphragm attached to the illuminator (the field stop) the condenser iris is adjusted for each objective
Staining• Staining is a biochemical technique of
adding a class-specific (DNA, proteins, lipids, carbohydrates) dye to a substrate to qualify or quantify the presence of a specific compound.
• Stains and dyes are frequently used in biology and medicine to highlight structures in biological tissues for viewing, often with the
aid of different microscopes.
DIC (Differential Interference Contrast)Wollaston Prism
Optical Path Length (OPL) = n • t
OPL difference = 2*pi*delta/lambda
delta=(n2 - n1) • t
`Nomarski`
Phase Contrast• Converts phase change to Amplitude change
http://micro.magnet.fsu.edu/primer/techniques/phasecontrast/phaseindex.html
• Converts phase change to Amplitude change
φ(x,y) < < 1
PSF(kx,ky) is the Point spread function (PSF)
Without PC optics
With PC optics
Furhter Contrast Enhancement in Phase Contrast Microscopy
• Select part of illumination
Reduce the size of the fat arrow