Journal ArticleDOI
Refractive-index-mismatch induced aberrations in single-photon and two-photon microscopy and the use of aberration correction.
Martin J. Booth,Tony Wilson +1 more
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TLDR
It is shown that, at large focusing depths, the resolution of 2-p conventional and 1-p confocal microscopes are almost identical, and an approximation based upon geometrical optics shows that the axial resolution of heavily aberrated PSFs is roughly proportional to focusing depth.Abstract:
We examine the effects of aberrations induced by a refrac- tive index mismatch on the signal level and resolution of single- photon (12p) and two-photon (22p), conventional and confocal scanning microscopes. In particular, we consider the aberrations in- troduced by an interface between oil/glass and water. Resolution is defined in terms of enclosed fluorescence, rather than full-width half- maximum, revealing more useful information for heavily aberrated point spread functions (PSFs). It is shown that, at large focusing depths, the resolution of 22p conventional and 12p confocal mi- croscopes are almost identical. The benefits of aberration correction are examined by removing Zernike aberration modes. With aberration correction, the best resolution is found for 12p confocal and 22p confocal modes. An approximation based upon geometrical optics is also introduced which shows that the axial resolution of heavily ab- errated PSFs is roughly proportional to focusing depth. © 2001 Society ofread more
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Journal ArticleDOI
Adaptive optics in microscopy
TL;DR: In this article, the sources of aberrations, their effects and their correction with adaptive optics, particularly in confocal and two-photon microscopes, are discussed. And applications of adaptive optics in the related areas of optical data storage, optical tweezers and micro/nanofabrication are also reviewed.
Book ChapterDOI
Two-photon photopolymerization and 3D lithographic microfabrication
Hong-Bo Sun,Satoshi Kawata +1 more
TL;DR: In this article, the authors give an overview of the development and current progress of femtosecond laser micro-nanofabrication based on multiphoton absorption, and particular emphasis is placed on two-photon photopolymerization.
Journal ArticleDOI
Confocal light sheet microscopy: micron-scale neuroanatomy of the entire mouse brain.
TL;DR: It is shown that CLSM allows reconstructing macroscopic brain volumes with sub-cellular resolution with a frame rate of 10 Hz, and the whole-brain high-resolution fluorescence imaging assured by CLSM may represent a powerful tool to navigate the brain through neuronal pathways.
Proceedings ArticleDOI
Adaptive optics in microscopy
TL;DR: Applications of adaptive optics in the related areas of optical data storage, optical tweezers and micro/nanofabrication are reviewed, particularly in confocal and two-photon microscopes.
Journal ArticleDOI
Characterizing specimen induced aberrations for high NA adaptive optical microscopy
TL;DR: The results indicate that adaptive correction of low order Zernike modes can provide significant benefit for many specimens and show that quantitative fluorescence microscopy may be strongly affected by specimen induced aberrations in non-adaptive systems.
References
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Journal ArticleDOI
Two-Photon Laser Scanning Fluorescence Microscopy
TL;DR: The fluorescence emission increased quadratically with the excitation intensity so that fluorescence and photo-bleaching were confined to the vicinity of the focal plane as expected for cooperative two-photon excitation.
BookDOI
Handbook of biological confocal microscopy
TL;DR: Methods for Three-Dimensional Imaging and Tutorial on Practical Confocal Microscopy and Use of the Confocal Test Specimen.
Journal ArticleDOI
Measurement of two-photon excitation cross sections of molecular fluorophores with data from 690 to 1050 nm
Chris Xu,Watt W. Webb +1 more
TL;DR: In this paper, the two-photon fluorescence excitation (TPE) spectra were measured for 11 common molecular fluorophores in the excitation wavelength range 690 nm < λ < 1050 nm.
Journal ArticleDOI
Multiphoton microscopy in life sciences
TL;DR: Owing to the high NIR penetration depth, non‐invasive optical biopsies can be obtained from patients and ex vivo tissue by morphological and functional fluorescence imaging of endogenous fluorophores such as NAD(P)H, flavin, lipofuscin, porphyrins, collagen and elastin.
Journal ArticleDOI
Theory and practice of scanning optical microscopy
Tony Wilson,Colin J. R. Sheppard +1 more