High speed functional imaging with source localized multifocal two-photon microscopy

Biomedical Optics Express
Peter QuickeAmanda J Foust


Multifocal two-photon microscopy (MTPM) increases imaging speed over single-focus scanning by parallelizing fluorescence excitation. The imaged fluorescence's susceptibility to crosstalk, however, severely degrades contrast in scattering tissue. Here we present a source-localized MTPM scheme optimized for high speed functional fluorescence imaging in scattering mammalian brain tissue. A rastered line array of beamlets excites fluorescence imaged with a complementary metal-oxide-semiconductor (CMOS) camera. We mitigate scattering-induced crosstalk by temporally oversampling the rastered image, generating grouped images with structured illumination, and applying Richardson-Lucy deconvolution to reassign scattered photons. Single images are then retrieved with a maximum intensity projection through the deconvolved image groups. This method increased image contrast at depths up to 112 μm in scattering brain tissue and reduced functional crosstalk between pixels during neuronal calcium imaging. Source-localization did not affect signal-to-noise ratio (SNR) in densely labeled tissue under our experimental conditions. SNR decreased at low frame rates in sparsely labeled tissue, with no effect at frame rates above 50 Hz. Our non-descan...Continue Reading


Apr 6, 1990·Science·W DenkW W Webb
Mar 10, 2001·Journal of Microscopy·T NielsenP Andresen
Jan 31, 2006·Journal of Neuroscience Methods·Rafael KurtzMarkus Sauer
Dec 19, 2007·Optics Letters·J BewersdorfS W Hell
Jan 1, 1995·IEEE Transactions on Image Processing : a Publication of the IEEE Signal Processing Society·J C YenS Chang
Jan 9, 2009·Frontiers in Neural Circuits·Volodymyr NikolenkoRafael Yuste
Sep 3, 2007·Optics Express·Ki Hean KimPeter T C So
Jan 11, 2011·Nature Methods·Adrian ChengCarlos Portera-Cailliau
Jul 8, 2011·Journal of Neurophysiology·Romain FranconvilleIsabel Llano
Apr 17, 2012·Journal of Biomedical Optics·Yonghong ShaoBruce Z Gao
Jun 27, 2013·Proceedings of the National Academy of Sciences of the United States of America·Mathieu DucrosSerge Charpak
Jan 11, 2014·The European Journal of Neuroscience·Mayumi TadaMasanobu Kano
Jun 6, 2014·Scientific Reports·Jae Won ChaPeter T C So
Jul 16, 2014·PeerJ·Stéfan van der Waltscikit-image contributors
Jan 22, 2015·Journal of Biological Methods·Arthur D EdelsteinNico Stuurman
Mar 18, 2015·Biomedical Optics Express·Peter RupprechtAlipasha Vaziri
Oct 29, 2015·Nature Reviews. Neuroscience·Artur LuczakKenneth D Harris
Nov 1, 2016·Nature Methods·Robert PrevedelAlipasha Vaziri
Dec 27, 2016·Biomedical Optics Express·Michael Lawrence CastanaresVincent R Daria
Jan 6, 2017·Scientific Reports·Serena BovettiTommaso Fellin
Nov 14, 2017·Journal of Neural Engineering·Renaud SchuckSimon R Schultz

Methods Mentioned

fluorescence imaging

Related Concepts

Diagnostic Imaging
Disease Susceptibility
Fluorescence Spectroscopy
Fluorescence Imaging
Brain Tissue

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