Simultaneous multilayer scanning and detection for multiphoton fluorescence microscopy.

Scientific Reports
Partha Pratim Mondal, Alberto Diaspro

Abstract

Fast three-(3D) imaging requires parallel optical slicing of a specimen with an efficient detection scheme. The generation of multiple localized dot-like excitation structures solves the problem of simultaneous slicing multiple specimen layers, but an efficient detection scheme is necessary. Confocal theta detection (detection at 90° to the optical axis) provides a suitable detection platform that is capable of cross-talk-free fluorescence detection from each nanodot (axial dimension ≈ 150 nm). Additionally, this technique has the unique feature of imaging a specimen at a large working distance with super-resolution capabilities. Polarization studies show distinct field structures for fixed and fluid samples, indicating a non-negligible field-dipole interaction. The realization of the proposed imaging technique will advance and diversify multiphoton fluorescence microscopy for numerous applications in nanobioimaging and optical engineering.

References

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Sep 6, 2005·Proceedings of the National Academy of Sciences of the United States of America·Mats G L Gustafsson
Feb 16, 2006·Quarterly Reviews of Biophysics·Alberto DiasproMaddalena Collini
Aug 7, 2007·Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics·Partha Pratim Mondal, Alberto Diaspro
Jan 18, 2008·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Jonas FöllingStefan W Hell
Sep 22, 2009·Methods in Molecular Biology·Alessandro EspositoAsparouh I Iliev
May 6, 2010·The Review of Scientific Instruments·Partha Pratim MondalAlberto Diaspro
Jul 19, 2011·Nature Methods·Thai V TruongScott E Fraser

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Citations

Jun 10, 2014·PloS One·Kavya MohanPartha Pratim Mondal
Jan 3, 2013·The Review of Scientific Instruments·Shilpa Dilipkumar, Partha Pratim Mondal
Nov 6, 2012·Trends in Molecular Medicine·Robert G EvansMichael Khan
Oct 19, 2013·Microscopy Research and Technique·Raju RegmiPartha P Mondal
Jun 12, 2016·Seminars in Cell & Developmental Biology·Gustavo de MedeirosLars Hufnagel
Sep 27, 2012·Scientific Reports·Subhajit B PurnapatraPartha Pratim Mondal
Aug 15, 2014·Optics Letters·Subhajit B PurnapatraPartha P Mondal
Jan 1, 2014·Frontiers in Molecular Biosciences·Partha Pratim Mondal
Feb 14, 2015·Optics Letters·Partha Pratim MondalKavya Mohan

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Methods Mentioned

BETA
fluorescence microscopy
fluorescence imaging
imaging technique
imaging techniques
optical
fluorescence correlation spectroscopy

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