Fast non-negative temporal deconvolution for laser scanning microscopy

J Biophotonics. 2013 Feb;6(2):153-62. doi: 10.1002/jbio.201100133. Epub 2012 Mar 21.

Abstract

Laser scanning microscopy (LSM) is a common technique for high resolution fluorescent imaging. Here we describe a fast algorithm for non-negative deconvolution and apply it to readout of LSM detector photocurrents. By broadening photon impulses and deconvolving sampled photocurrent, effective quantum efficiency of the imaging system is increased. Using simulation and imaging with a custom-built two-photon microscope, we demonstrate improved fidelity of images acquired at short dwell times over a wide range of photon rates. Images formed show increased correlation-to-sample equivalent to a 25% increase in photon rate, lower noise, and reduced bleed-through compared to conventional image generation.

MeSH terms

  • Algorithms
  • Animals
  • Calcium Signaling
  • Computer Simulation
  • Fluorescent Dyes
  • Microscopy, Confocal / methods*
  • Microscopy, Confocal / statistics & numerical data
  • Microscopy, Fluorescence, Multiphoton / methods
  • Microscopy, Fluorescence, Multiphoton / statistics & numerical data
  • Molecular Imaging
  • Neurons / metabolism
  • Optical Phenomena
  • Signal Processing, Computer-Assisted
  • Superior Colliculi / metabolism
  • Xenopus laevis

Substances

  • Fluorescent Dyes