Fig. 2.

A: Point sources of light are significantly blurred by both the point spread function (PSF) of the microscope and noise. Two distinct methods exist for post-acquisition processing of a blurred image. Deblurring algorithms remove out of focus planes, producing a sharper image but reducing the total information available and thus rendering the image unquantifiable. Deconvolution algorithms preserve the information, and restore the blurred fluorescence to the original point source using a range of methods as described in the text. B,C: Cohesin forms a barrel-like structure during metaphase in budding yeast, which appears as a doughnut when viewed end-on and bilobed when viewed side-on (Yeh et al., 2008; Stephens et al., 2011). Original and deconvolved images of cohesin (SMC3-GFP) illustrating how both input image quality and deconvolution algorithm parameters can lead to very different output images and potentially introduce artifacts.