Intravital Two-Photon Excitation Microscopy in Neuroscience: General Concepts and Applications
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Two-photon excitation microscopy
Intravital microscopy
Summary We briefly describe the advantages and limitations of label‐free multiphoton microscopy and probe‐labeled two‐photon microscopy for the endomicroscopic diagnosis. The two methods are complementary and more information can be collected from tissues by combining the two methods. Therefore, parallel efforts should be directed to the development of both label‐free MPM and probe‐labeled TPM as the diagnostic tool. SCANNING 36:462–464, 2014. © 2013 Wiley Periodicals, Inc.
Two-photon excitation microscopy
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Two-photon excitation microscopy (also referred to as multiphoton laser scanning microscopy) has gained increasing popularity during the past few years because of the distinct advantages over single-photon microscopy, which includes increased penetration depth and low out-of-focus photobleaching and photodamage. It allows superior imaging of thick specimen compared to single-photon microscopy, and it excels at imaging live cells either single or within intact tissue. This highly valuable tool has been used with great success to gain important new insights into brain tissue, embryos, whole organs, entire animals, and it has been most useful in numerous other applications.
Photobleaching
Two-photon excitation microscopy
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In this talk a brief review on the non linear laser imaging techniques will be displayed. In particular, two photon fluorescence microscopy, lifetime imaging, multispectral imaging, second harmonic generation microscopy principles will be described.
Two-photon excitation microscopy
Fluorescence-lifetime imaging microscopy
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The structural origin of signals in multi-photon microscopy of biological tissues is determined. The combined use of two-photon excited fluorescence and second- harmonic generation in reflection geometry provides complementary information that allows non-invasive tissue characterization.
Characterization
Two-photon excitation microscopy
Reflection
Biological Imaging
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Intravital microscopy
Two-photon excitation microscopy
Fluorescence-lifetime imaging microscopy
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Multiphoton microscopy is a powerful technique based on complex quantum mechanical effects. Thanks to the development of turnkey mode-locked laser systems, multiphoton microscopy is now available for everyone to use without extreme complexity. In this short introduction, we describe qualitatively the important concepts underlying the most commonly used type of multiphoton microscopy (two-photon excitation). We elucidate how those properties lead to the powerful results that have been achieved using this technique. As with any technique, two-photon excitation microscopy has limitations that we describe, and we provide examples of particular classes of experiments where two-photon excitation microscopy is advantageous over other approaches. Finally, we briefly describe other useful multiphoton microscopy approaches, such as three-photon excitation and second harmonic generation imaging.
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Two-photon excitation microscopy
Fluorescence-lifetime imaging microscopy
Photoactivated localization microscopy
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Intravital microscopy
Ex vivo
Two-photon excitation microscopy
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Abstract Two‐photon fluorescence microscopy allows three‐dimensional imaging of biological specimens in vivo . Compared with confocal microscopy, it offers the advantages of deeper tissue penetration and less photodamage but has the disadvantage of slightly lower resolution.
Two-photon excitation microscopy
Photoactivated localization microscopy
Fluorescence-lifetime imaging microscopy
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Efficient two-photon (2PA) absorbing dyes and bioconjugates were used in two-photon fluorescence microscopy (2PFM) of cells, tissue sections, and excised tumors. Results show the utility of these dyes in studying biological processes.
Two-photon excitation microscopy
Ex vivo
Fluorescence-lifetime imaging microscopy
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