Adaptive aberration correction in a two‐photon microscope

Neil, Mark A. A.; Juškaitis, Rimas; Booth, Martin J.; Wilson, Tony; Tanaka, Tomonari; Kawata, Satoshi

doi:10.1046/j.1365-2818.2000.00770.x

Cited by 144 publications

(119 citation statements)

References 21 publications

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“…Second, MPM will be much more sensitive to any factor in tissue that broadens the focused spot. Consistent with this, studies in model systems have shown MPM is highly sensitive to spherical aberration [12,22,24,25].…”

Section: The Role Of Spherical Aberration In Deep-tissue Multiphoton supporting

confidence: 55%

“…Additional methods for manipulating spherical aberration, including use of different immersion media, different thickness coverslips, and adaptive lens systems, may increase the range of tissues and depths that can be accommodated by spherical aberration compensation. The most complete (and most elaborate) solution to the problem of aberration of the focus may lie in adaptive optics systems, which have been demonstrated to improve resolution and signal level in refractive index mismatched samples [22,25] and in living tissue [37]. Effect of correction collar adjustment on fluorescence intensity as a function of depth in kidney tissue.…”

Section: Resultsmentioning

confidence: 99%

“…In addition, signal levels can decline with depth when the refractive index of the tissue is not matched to that of the immersion fluid of the objective. [22][23][24][25] This "refractive index mismatch" results in spherical aberration, a condition in which paraxial light rays and peripheral light rays focus to different places, broadening the focal point of the objective lens. While both confocal and multiphoton microscopy are susceptible to each of these factors, the magnitude of their effects is much reduced by the unique design of the multiphoton microscope, as described below.…”

Section: Signal Attenuation In Multiphoton Microscopymentioning

confidence: 99%

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Improving signal levels in intravital multiphoton microscopy using an objective correction collar

Muriello

Dunn

2008

Optics Communications

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Multiphoton microscopy has enabled biologists to collect high-resolution images hundreds of microns into biological tissues, including tissues of living animals. While the depth of imaging exceeds that possible from any other form of light microscopy, multiphoton microscopy is nonetheless generally limited to depths of less than a millimeter. Many of the advantages of multiphoton microscopy for deep tissue imaging accrue from the unique nature of multiphoton fluorescence excitation. However, the quadratic relationship between illumination level and fluorescence excitation makes multiphoton microscopy especially susceptible to factors that degrade the illumination focus. Here we examine the effect of spherical aberration on multiphoton microscopy in fixed kidney tissues and in the kidneys of living animals. We find that spherical aberration, as evaluated from axial asymmetry in the point spread function, can be corrected by adjustment of the correction collar of a water immersion objective lens. Introducing a compensatory positive spherical aberration into the imaging system decreased the depth-dependence of signal levels in images collected from living animals, increasing signal by up to 50%.

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Section: The Role Of Spherical Aberration In Deep-tissue Multiphoton supporting

confidence: 55%

Section: Resultsmentioning

confidence: 99%

Section: Signal Attenuation In Multiphoton Microscopymentioning

confidence: 99%

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Improving signal levels in intravital multiphoton microscopy using an objective correction collar

Muriello

Dunn

2008

Optics Communications

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“…Characteristic of PAMs are the absence of macroscopic moving parts and software control of the (arbitrary) patterns of illumination and detection. Thus, the SLM may be programmed for very diverse requirements, such as in endoscopy (5,17), profilometry (18,19), point spread function engineering (20), adaptive correction of aberrations (21), and photodirected oligonucleotide synthesis (22).…”

Section: Analytical Cytologymentioning

confidence: 99%

Selective photoreactions in a programmable array microscope (PAM): Photoinitiated polymerization, photodecaging, and photochromic conversion

et al. 2005

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BackgroundInnovative thinking and experimentation were the hallmarks of Mack Fulwyler's approach to research. This report summarizes some of the ideas and their early realizations that he pursued in the field of imaging cytometry, work that was not published before his untimely death, although he composed the initial draft of this report.MethodsIncluded are related experiments implemented in the programmable array microscope (PAM) devised for patterned illumination and detection, the instrument that Mack Fulwyler employed during a sabbatical leave in Göttingen in 1998. Despite being the originator of instrumentation for flow cytometry and sorting, Mack Fulwyler was intensely interested in imaging systems, recognizing their ability to resolve cellular details obscured by the whole cell signals generally acquired in flow. At one point, these interests merged with those of two other authors (I.T.Y. and T.M.J.), leading to the Image Cytometry and Sorting (ICAS) strategy and project. A major goal was uncomplicated rare cell detection and isolation using a sequential process of cellular labeling via suitable probes, whole field imaging, and selective area‐restricted photoinduced reactions designed to encapsulate and/or chemically or physically tag cells in a manner permitting subsequent fractionation by bulk techniques.Results and ConclusionThis publication features photoinduced polymerization, photodecaging, photoactivation, and photochromic conversion reactions carried out by Fulwyler and/or the other authors with the PAM, employing operator designated patterns and locations in various samples. Photopolymerization of polyethylene glycol‐diacrylate to a gel‐like structure allowing the specific selection of objects (cells) for further analysis and processing techniques was the approach explored personally by Mack Fulwyler in relation to the ICAS concept. © 2005 International Society for Analytical Cytology

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“…4 In microscopy, significant progress has been made in confocal systems, 5,6 and adaptive optics has also been applied to nonlinear microscopies such as two-photon excitation microscopy, 7,8 harmonic generation microscopy, 9,10 and coherent anti-stokes Raman scattering ͑CARS͒ microscopy. 11 The different implementations of adaptive optics to nonlinear microscopy can be roughly classified into two approaches: feedback via two-photon excited fluorescence light, or feedback via reflected excitation light.…”

Section: Introductionmentioning

confidence: 99%

A Shack-Hartmann Wavefront Sensor Based Adaptive Optics System for Multiphoton Microscopy

Won

2008

Biomedical Optics

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Abstract. The imaging depth of two-photon excitation fluorescence microscopy is partly limited by the inhomogeneity of the refractive index in biological specimens. This inhomogeneity results in a distortion of the wavefront of the excitation light. This wavefront distortion results in image resolution degradation and lower signal level. Using an adaptive optics system consisting of a Shack-Hartmann wavefront sensor and a deformable mirror, wavefront distortion can be measured and corrected. With adaptive optics compensation, we demonstrate that the resolution and signal level can be better preserved at greater imaging depth in a variety of ex-vivo tissue specimens including mouse tongue muscle, heart muscle, and brain. However, for these highly scattering tissues, we find signal degradation due to scattering to be a more dominant factor than aberration.

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Adaptive aberration correction in a two‐photon microscope

Cited by 144 publications

References 21 publications

Improving signal levels in intravital multiphoton microscopy using an objective correction collar

Improving signal levels in intravital multiphoton microscopy using an objective correction collar

Selective photoreactions in a programmable array microscope (PAM): Photoinitiated polymerization, photodecaging, and photochromic conversion

A Shack-Hartmann Wavefront Sensor Based Adaptive Optics System for Multiphoton Microscopy

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