In Vivo Multiphoton Microscopy of Deep Brain Tissue

Levene, Michael J.; Dombeck, Daniel A.; Kasischke, Karl A.; Molloy, Raymond; Webb, Watt W.

doi:10.1152/jn.01007.2003

Cited by 423 publications

(364 citation statements)

References 22 publications

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“…The introduction of multiphoton microscopy promises significant progress in in situ even in vivo investigation of tissue microcomponent with high precision and contrast (Brakenhoff et al, 1996;König et al, 1996;Zipfel et al, 2003a;Levene et al, 2004;Diaspro et al, 2005;Wang et al, 2005b). Based on this technique, visualization of the intracellular mitochondria (Barzda et al, 2005) and quantification of intracellular serotonin (Maiti et al, 1997) and NAD(P)H (Patterson et al, 2000) have been reported.…”

Section: Corneal 2-photon Excitation Imagingmentioning

confidence: 99%

“…Dendrites and spines can be imaged to study the alteration in their shape and size. The changes in axonal morphology of deep brain tissues were acquired through imaging of the activity of neuronal populations (Brecht et al, 2004;Levene et al, 2004;Nitsch et al, 2004;Chia & Levene, 2009). In particular, the cortical astrocytes were imaged in the intact mouse brain (Tian et al, 2006).…”

Section: Further Uses Of Multiphoton Excitation Imaging In Tumourous mentioning

confidence: 99%

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Two‐photon microscopy of deep intravital tissues and its merits in clinical research

Wang

König

2010

Journal of Microscopy

178

131

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SummaryMultiphoton excitation laser scanning microscopy, relying on the simultaneous absorption of two or more photons by a molecule, is one of the most exciting recent developments in biomedical imaging. Thanks to its superior imaging capability of deeper tissue penetration and efficient light detection, this system becomes more and more an inspiring tool for intravital bulk tissue imaging. Two-photon excitation microscopy including 2-photon fluorescence and second harmonic generated signal microscopy is the most common multiphoton microscopic application. In the present review we take diverse ocular tissues as intravital samples to demonstrate the advantages of this approach. Experiments with registration of intracellular 2-photon fluorescence and extracellular collagen second harmonic generated signal microscopy in native ocular tissues are focused. Data show that the in-tandem combination of 2-photon fluorescence and second harmonic generated signal microscopy as two-modality microscopy allows for in situ co-localization imaging of various microstructural components in the whole-mount deep intravital tissues. New applications and recent developments of this high technology in clinical studies such as 2-photon-controlled drug release, in vivo drug screening and administration in skin and kidney, as well as its uses in tumourous tissues such as melanoma and glioma, in diseased lung, brain and heart are additionally reviewed. Intrinsic emission two-modal 2-photon microscopy/tomography, acting as an efficient and sensitive non-injurious imaging approach featured by high contrast and subcellular spatial resolution, has been proved to be a promising tool for intravital deep tissue imaging and clinical studies. Given the level of its performance, we believe Correspondence to B.-G. Wang. Tel: +49 3641 938496; fax: +49 3641 938552; e-mail: Baogui.Wang@mti.uni-jena.de that the non-linear optical imaging technique has tremendous potentials to find more applications in biomedical fundamental and clinical research in the near future.

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Section: Corneal 2-photon Excitation Imagingmentioning

confidence: 99%

Section: Further Uses Of Multiphoton Excitation Imaging In Tumourous mentioning

confidence: 99%

Two‐photon microscopy of deep intravital tissues and its merits in clinical research

Wang

König

2010

Journal of Microscopy

178

131

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“…Like other multi-photon techniques [10][11][12][13][14][15][16][17][18], THG not only enables the recording of label-free images of unfixed 3D volumes of tissue [1][2][3][4][5][6][7][8][9][10] free from spatial distortion artifacts inherent to histopathology, but also potentially allows feedback on the nature of the tissue, i. e. whether it is healthy or tumorous, to the surgeon during surgery, as the relative speed of the imaging modalities approaches 'real' time, and no preparation steps of the tissue are required [5,6,[19][20][21][22][23]. THG was shown recently to yield label-free images of ex-vivo human tumor tissue of histopathological quality, in real-time [6].…”

Section: Introductionmentioning

confidence: 99%

Quantitative comparison of 3D third harmonic generation and fluorescence microscopy images

Zhang

Kuzmin

Groot

et al. 2017

Journal of Biophotonics

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Third harmonic generation (THG) microscopy is a label-free imaging technique that shows great potential for rapid pathology of brain tissue during brain tumor surgery. However, the interpretation of THG brain images should be quantitatively linked to images of more standard imaging techniques, which so far has been done qualitatively only. We establish here such a quantitative link between THG images of mouse brain tissue and all-nuclei-highlighted fluorescence images, acquired simultaneously from the same tissue area. For quantitative comparison of a substantial pair of images, we present here a segmentation workflow that is applicable for both THG and fluorescence images, with a precision of 91.3 % and 95.8 % achieved respectively. We find that the correspondence between the main features of the two imaging modalities amounts to 88.9 %, providing quantitative evidence of the interpretation of dark holes as brain cells. Moreover, 80 % bright objects in THG images overlap with nuclei highlighted in the fluorescence images, and they are 2 times smaller than the dark holes, showing that cells of different morphologies can be recognized in THG images. We expect that the described quantitative comparison is applicable to other types of brain tissue and with more specific staining experiments for cell type identification.

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“…Several studies have shown that multiple-spectra QDs can be used to achieve coincident tracking of multiple tumor cells in vivo using fluorescence emission-scanning microscopy (Gao et al, 2004;. In the central nervous system, microangiography of deep brain blood vessels and nonspecific labeling of brain cells has been achieved following QD injection into arterial blood (Levene et al, 2004;Voura et al, 2004).…”

Section: Introductionmentioning

confidence: 99%