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Chapter 4 –Microscopy

Chapter 4 Microscopy Gabriel PopescuUniversity of Illinois at Urbana ChampaignypgBeckman InstituteQuantitativeLightImagingLaborat oryElectrical and Computer Engineering, UIUCP rinciples of Optical ImagingQuantitative Light Imaging 460 Optical Imaging The Microscope can be approximated by 2 lenses [ [ [ , ]]] Resolution of Optical MicroscopespppyTube lensSampleF2 F2 Objective[ [ [ , ]]]yU(x,y)F 1[[ ]]FUxy[[[ ]]] []xyFFUU sign means inverted image Theobjectiveisthemostimportantpartofthem icroscope1[[,]]FUxy21[[[,]]][,]yFFUxyUMM The objective is the most important part of the microscope Usually a third lens (ocular) images F2 at , such that we can visualize it with the relaxed 4: Microscopy 460 Optical Imaging The objective lens dictates the resolution or size of the Resolution of Optical Microscopesjsmallest object that the microscope can resolve. Contrast is generated by absorption, scattering, etc. Microscopes can be categorized by the methods that they use to produce contrast.

focal distance measured from principal plane working distance = distance from F to physical surface of lens Entrance Pupil image of physical aperture f W Entrance Pupil = image of physical apert and entrance pupil determine numerical aperf ture i.e. resolution Chapter 4: Microscopy 10

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