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Fast non‐negative temporal deconvolution for laser scanning microscopy
Author(s) -
Podgorski Kaspar,
Haas Kurt
Publication year - 2013
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201100133
Subject(s) - deconvolution , microscopy , optics , detector , dwell time , two photon excitation microscopy , materials science , laser , photon , resolution (logic) , physics , microscope , fluorescence , computer science , artificial intelligence , medicine , clinical psychology
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. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)