&lt;title&gt;Suitable microscope objectives for multiphoton digital imaging&lt;/title&gt;

Benham, Gerald S.; Schwartz, Stanley

doi:10.1117/12.470673

Cited by 3 publications

(3 citation statements)

References 11 publications

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“…Then, the excitation light should reach the microscope objective passing through the minimum number of optical components and possibly along the shortest path. Typically, high NA objectives, with high IR transmission (70-80 %), are used to maximize 2PE efficiency (Benham, 2002 ;Benham & Schwartz, 2002). While the x-y scanners provide lateral focal-point scanning, axial scanning can be achieved by means of different positioning devices, the most popular being: belt-driven systems using a DC motor and objective piezo nano-positioners (Diaspro, 2001b).…”

Section: General Aspectsmentioning

confidence: 99%

“…For a fluorescence microscope, special criteria have to be satisfied (Benham, 2002 ;Benham & Schwartz, 2002) for the choice of the objective lens besides those common to any microscope (Keller, 1995). For 2PE, an adequate transmission in the IR regime has to be coupled with good collection efficiency towards the UV region.…”

Section: Lens Objectives and Pulse Broadeningmentioning

confidence: 99%

“…These factors play a large role in determining whether a particular sample can be successfully imaged, and in particular on the maximum depth that can be visualized (Girkin & Wokosin, 2001a ;Jonkman & Stelzer, 2001 ;de Grauw & Gerritsen, 2001). Table 4 shows a collection of transmission data for Nikon CFI60 series of objectives (Girkin & Wokosin, 2001a ;Benham & Schwartz, 2002).…”

Section: Lens Objectives and Pulse Broadeningmentioning

confidence: 99%

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Two-photon fluorescence excitation and related techniques in biological microscopy

Diaspro

Chirico

Collini

2005

Quart. Rev. Biophys.

281

217

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This review is concerned with two-photon excited fluorescence microscopy (2PE) and related techniques, which are probably the most important advance in optical microscopy of biological specimens since the introduction of confocal imaging. The advent of 2PE on the scene allowed the design and performance of many unimaginable biological studies from the single cell to the tissue level, and even to whole animals, at a resolution ranging from the classical hundreds of nanometres to the single molecule size. Moreover, 2PE enabled long-term imaging of in vivo biological specimens, image generation from deeper tissue depth, and higher signal-to-noise images compared to wide-field and confocal schemes. However, due to the fact that up to this time 2PE can only be considered to be in its infancy, the advantages over other techniques are still being evaluated. Here, after a brief historical introduction, we focus on the basic principles of 2PE including fluorescence correlation spectroscopy. The major advantages and drawbacks of 2PE-based experimental approaches are discussed and compared to the conventional single-photon excitation cases. In particular we deal with the fluorescence brightness of most used dyes and proteins under 2PE conditions, on the optical consequences of 2PE, and the saturation effects in 2PE that mostly limit the fluorescence output. A complete section is devoted to the discussion of 2PE of fluorescent probes. We then offer a description of the central experimental issues, namely: choice of microscope objectives, two-photon excitable dyes and fluorescent proteins, choice of laser sources, and effect of the optics on 2PE sensitivity. An inevitably partial, but vast, overview of the applications and a large and up-to-date bibliography terminate the review. As a conclusive comment, we believe that 2PE and related techniques can be considered as a mainstay of the modern biophysical research milieu and a bright perspective in optical microscopy.

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Section: General Aspectsmentioning

confidence: 99%

Section: Lens Objectives and Pulse Broadeningmentioning

confidence: 99%

Section: Lens Objectives and Pulse Broadeningmentioning

confidence: 99%

See 1 more Smart Citation

Two-photon fluorescence excitation and related techniques in biological microscopy

Diaspro

Chirico

Collini

2005

Quart. Rev. Biophys.

281

217

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Two-Photon Excitation Fluorescence Microscopy

Diaspro

Schneider²,

Bianchini

et al. 2007

Science of Microscopy

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A pragmatic guide to multiphoton microscope design

et al. 2015

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Multiphoton microscopy has emerged as a ubiquitous tool for studying microscopic structure and function across a broad range of disciplines. As such, the intent of this paper is to present a comprehensive resource for the construction and performance evaluation of a multiphoton microscope that will be understandable to the broad range of scientific fields that presently exploit, or wish to begin exploiting, this powerful technology. With this in mind, we have developed a guide to aid in the design of a multiphoton microscope. We discuss source selection, optical management of dispersion, image-relay systems with scan optics, objective-lens selection, single-element light-collection theory, photon-counting detection, image rendering, and finally, an illustrated guide for building an example microscope.

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<title>Suitable microscope objectives for multiphoton digital imaging</title>

Cited by 3 publications

References 11 publications

Two-photon fluorescence excitation and related techniques in biological microscopy

Two-photon fluorescence excitation and related techniques in biological microscopy

Two-Photon Excitation Fluorescence Microscopy

A pragmatic guide to multiphoton microscope design

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