Sampling How does digital sampling affect resolution Look at imaging these object with a digital camera How close together do pixels need to be? Sampling over time: How often do you need to image a moving sample Nyquist theory states that you should sample more than 2 X the frequency that you expect. Over sampling Nyquist sampling Under sampling causes aliasing Optical Transfer Function MTF When objects get close together the contrast decreases. MTF = Image Modulation/Object Modulation MTF = 2(φ - cosφsinφ)/π   and φ = cos-1(λν/2NA) The Optical Transfer function is the Modulation transfer function times a phase component. OTF = MTF × eiφ(f) Camera bit depth 0 or 1 00 or 01 or 10 or 11 000 or 001 or 010 or 011 or 100 or 101 or 110 or 111 and so on Jpg is 8 bit Tiff can be 16 bit references https://imb.uq.edu.au/research/facilities/microscopy/training-manuals/microscopy-online-resources/image-capture/nyquist-conditions https://microscopy.berkeley.edu/courses/dib/sections/02images/sampling.html https://ocw.mit.edu/courses/mechanical-engineering/2-71-optics-spring-2009/video-lectures/lecture-22-coherent-and-incoherent-imaging/MIT2_71S09_lec22.pdf