Microscopy Resolution and Object Distinction
Learn about the concept of microscopy resolution, the factors affecting it, and how objects can be distinguished based on resolving power. Discover the importance of numerical aperture and refractive index in achieving sharper images with different objective lenses.
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Presentation Transcript
Microscopy Resolution = Resolving Power -the smallest distance (d) at which two objects can be successfully distinguished. Resolution (d): d = (0.61 x )/ NA = wave length NA= numerical aperture = of angular aperture n = refractive index of medium NA= n sin
NA= n sin Refractive index ( ) of different media Air=1.0003 Water=1.33 Immersion Oil=1.515
Resolution versus Wavelength Resolution: d= 0.61 x NA Wavelength (nanometers) 360 400 450 500 550 600 650 700 Resolution (micrometers) .19 .21 .24 .26 .29 .32 .34 .37
Resolving Distance (d) Human eye Light Microscope Scanning Electron Microscope Transmission Electron Microscope 2.5 nm 1.0 nm 0.2 mm 0.2 um Resolution: d= (0.61 x )/ NA HistoTip: Avoid confusion when discussing resolution. Increased resolution or resolving power usually means a SMALLER value of d (distance).
PROBLEM: Objective lens A: Magnification = 40X N.A. = 0.45 Objective lens B: Magnification = 40X N.A. = 0.80 -->Which objective lens would give the sharper image and why?
