Three-dimensional imaging by deconvolution microscopy

James G. McNally, Tatiana Karpova, John Cooper, José Angel Conchello

Research output: Contribution to journalArticlepeer-review

375 Scopus citations


Deconvolution is a computational method used to reduce out-of-focus fluorescence in three-dimensional (3D) microscope images. It can be applied in principle to any type of microscope image but has most often been used to improve images from conventional fluorescence microscopes. Compared to other forms of 3D light microscopy, like confocal microscopy, the advantage of deconvolution microscopy is that it can be accomplished at very low light levels, thus enabling multiple focal-plane imaging of light-sensitive living specimens over long time periods. Here we discuss the principles of deconvolution microscopy, describe different computational approaches for deconvolution, and discuss interpretation of deconvolved images with a particular emphasis on what artifacts may arise.

Original languageEnglish
Pages (from-to)373-385
Number of pages13
JournalMethods: A Companion to Methods in Enzymology
Issue number3
StatePublished - Nov 1999


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