Application of multiline two-photon microscopy to functional in vivo imaging

Kurtz, Rafael; Fricke, Marc; Kalb, Julia; Tinnefeld, Philip; Sauer, Markus

doi:10.1016/j.jneumeth.2005.12.003

Cited by 59 publications

(47 citation statements)

References 44 publications

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“…The third method increases imaging speed by parallelizing the multiphoton imaging process and has been called multifocal multiphoton microscopy (MMM) [20][21][22][23][24]. In MMM, a specimen is scanned with multiple excitation foci instead of a single excitation focus.…”

Section: Introductionmentioning

confidence: 99%

Multifocal multiphoton microscopy based on multianode photomultiplier tubes

Kim¹,

Buehler²,

Bahlmann³

et al. 2007

Opt. Express

105

114

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Multifocal multiphoton microscopy (MMM) enhances imaging speed by parallelization. It is not well understood why the imaging depth of MMM is significantly shorter than conventional singlefocus multiphoton microscopy (SMM). In this report, we show that the need for spatially resolved detectors in MMM results in a system that is more sensitive to the scattering of emission photons with reduced imaging depth. For imaging depths down to twice the scattering mean free path length of emission photons ( ), the emission point spread function (PSF em ) is found to consist of a narrow, diffraction limited distribution from ballistic emission photons and a broad, relatively low amplitude distribution from scattered photons. Since the scattered photon distribution is approximately 100 times wider than that of the unscattered photons at , image contrast and depth are degraded without compromising resolution. To overcome the imaging depth limitation of MMM, we present a new design that replaces CCD cameras with multi-anode photomultiplier tubes (MAPMTs) allowing more efficient collection of scattered emission photons. We demonstrate that MAPMT-based MMM has imaging depth comparable to SMM with equivalent sensitivity by imaging tissue phantoms, ex vivo human skin specimens based on endogenous fluorophores, and green fluorescent protein (GFP) expressing neurons in mouse brain slices.

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Section: Introductionmentioning

confidence: 99%

Multifocal multiphoton microscopy based on multianode photomultiplier tubes

Kim¹,

Buehler²,

Bahlmann³

et al. 2007

Opt. Express

105

114

View full text Add to dashboard Cite

show abstract

“…Using acousto-optic de°ectors for fast point scanning, however, has provided limited improvement due to signi¯cant dispersion of ultrashort pulses and spherical aberrations introduced by the optical elements used. 139 In MMM, the sample is scanned with multiple excitation focus points, [134][135][136] or lines 140 rather than a single excitation focus. The emitted light is collected by the multiple foci on a charge-coupled device (CCD) camera.…”

Section: Multifocal Multiphoton Microscopymentioning

confidence: 99%

Advanced optical microscopy methods for in vivo imaging of sub-cellular structures in thick biological tissues

Chen

Rehman

Sheppard

2014

J. Innov. Opt. Health Sci.

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Optical microscopy has become an indispensable tool for visualizing sub-cellular structures and biological processes. However, scattering in biological tissues is a major obstacle that prevents high-resolution images from being obtained from deep regions of tissue. We review common techniques, such as multiphoton microscopy (MPM) and optical coherence microscopy (OCM), for di®raction limited imaging beyond an imaging depth of 0.5 mm. Novel implementations have been emerging in recent years giving higher imaging speed, deeper penetration, and better image quality. Focal modulation microscopy (FMM) is a novel method that combines confocal spatial ltering with focal modulation to reject out-of-focus background. FMM has demonstrated an imaging depth comparable to those of MPM and OCM, near-real-time image acquisition, and the capability for multiple contrast mechanisms.

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“…Novel high-speed two-photon microscopes 22,35 enable image acquisition at video rates and will therefore be less hampered by image distortion while providing good SNRs. Still, triggered acquisition is required to limit between-frame motion artifacts.…”

Section: Journal Of Biomedical Opticsmentioning

confidence: 99%

In vivo high-resolution structural imaging of large arteries in small rodents using two-photon laser scanning microscopy

et al. 2010

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Document VersionPublisher's PDF, also known as Version of Record (includes final page, issue and volume numbers) Please check the document version of this publication:• A submitted manuscript is the author's version of the article upon submission and before peer-review. There can be important differences between the submitted version and the official published version of record. People interested in the research are advised to contact the author for the final version of the publication, or visit the DOI to the publisher's website.• The final author version and the galley proof are versions of the publication after peer review.• The final published version features the final layout of the paper including the volume, issue and page numbers. Link to publicationCitation for published version (APA): Megens, R. T. A., Reitsma, S., Prinzen, L., Oude Egbrink, M. G. A., Engels, W., Leenders, P. J. A., ... Zandvoort, van, M. (2010). In vivo high-resolution structural imaging of large arteries in small rodents using twophoton laser scanning microscopy. Journal of Biomedical Optics, 15(1), 011108-1/10. DOI: 10.1117/1.3281672 General rightsCopyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.• Users may download and print one copy of any publication from the public portal for the purpose of private study or research.• You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal ? Take down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Abstract. In vivo ͑molecular͒ imaging of the vessel wall of large arteries at subcellular resolution is crucial for unraveling vascular pathophysiology. We previously showed the applicability of two-photon laser scanning microscopy ͑TPLSM͒ in mounted arteries ex vivo. However, in vivo TPLSM has thus far suffered from in-frame and betweenframe motion artifacts due to arterial movement with cardiac and respiratory activity. Now, motion artifacts are suppressed by accelerated image acquisition triggered on cardiac and respiratory activity. In vivo TPLSM is performed on rat renal and mouse carotid arteries, both surgically exposed and labeled fluorescently ͑cell nuclei, elastin, and collagen͒. The use of short acquisition times consistently limit in-frame motion artifacts. Additionally, triggered imaging reduces between-frame artifacts. Indeed, structures in the vessel wall ͑cell nuclei, elastic laminae͒ can be imaged at subcellular resolution. In mechanically damaged carotid arteries, even the subendothelial collagen sheet ͑ϳ1 m͒ is visualized using collagentargeted quantum dots. We demonstrate stable in vivo imaging of large...

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Application of multiline two-photon microscopy to functional in vivo imaging

Cited by 59 publications

References 44 publications

Multifocal multiphoton microscopy based on multianode photomultiplier tubes

Multifocal multiphoton microscopy based on multianode photomultiplier tubes

Advanced optical microscopy methods for in vivo imaging of sub-cellular structures in thick biological tissues

In vivo high-resolution structural imaging of large arteries in small rodents using two-photon laser scanning microscopy

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