Enhanced three-dimensional deconvolution microscopy using a measured depth-varying point-spread function

Joshua W. Shaevitz, Daniel A. Fletcher

Research output: Contribution to journalArticlepeer-review

47 Scopus citations


We present a technique to systematically measure the change in the blurring function of an optical microscope with distance between the source and the coverglass (the depth) and demonstrate its utility in three-dimensional (3D) deconvolution. By controlling the axial positions of the microscope stage and an optically trapped bead independently, we can record the 3D blurring function at different depths. We find that the peak intensity collected from a single bead decreases with depth and that the width of the axial, but not the lateral, profile increases with depth. We present simple convolution and deconvolution algorithms that use the full depth-varying point-spread functions and use these to demonstrate a reduction of elongation artifacts in a reconstructed image of a 2 μm sphere.

Original languageEnglish (US)
Pages (from-to)2622-2627
Number of pages6
JournalJournal of the Optical Society of America A: Optics and Image Science, and Vision
Issue number9
StatePublished - Sep 2007

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics
  • Computer Vision and Pattern Recognition


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