Name: thinned-skull cortical window technique for in vivo optical coherence tomography imaging
Uploaded: 2012-11-19T13:00:00
Description: Watch this Scientific Journal Video about Thinned-skull Cortical Window Technique for In Vivo Optical Coherence Tomography Imaging at JoVE.com

Dette arbejde blev støttet af UC Discovery Proof of Concept tilskud og af NIH (R00 EB007241). Forfatterne vil også gerne takke Jacqueline Hubbard for hendes hjælp i dette eksperiment.

1. Kirurgisk forberedelse <ol><li> Female CD 1 mus i alderen 6-8 uger blev anvendt i vore forsøg. </li><li> Bedøve mus med en intraperitoneal injektion af en ketamin og xylazin blanding (80 mg / kg ketamine/10 mg / kg xylazin). Anbring musen på en homeothermic pad til at sikre en optimal kropstemperatur ved ~ 37 ° C. Løbende overvåge niveau af anæstesi ved at teste dyrets reflekser (f.eks knibe fod med stumpe pincet) og injicere mere anæstesi når det er nødvendigt. </li><li> Smør begge øjne med en...

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	<li>Bizheva, K., Unterhuber, A., Hermann, B., Povazay, B., Sattmann, H., Drexler, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+ex+vivo+and+in+vitro+brain+morphology+in+animal+models+with+ultrahigh+resolution+optical+coherence+tomography.">Imaging ex vivo and in vitro brain morphology in animal models with ultrahigh resolution optical coherence tomography.</a> Journal of Biomedical Optics. 9, 719-724 (2004).</li><li>Fujimoto, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+coherence+tomography+for+ultrahigh+resolution+in+vivo+imaging.">Optical coherence tomography for ultrahigh resolution in vivo imaging.</a> Nature Biotechnology. 21, 1361-1367 (2003).</li><li>Wantanabe, H., Rajagopalan, U. M., Nakamichi, Y., Igarashi, K. M., Kadono, H., Tanifuji, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Swept+source+optical+coherence+tomography+as+a+tool+for+real+time+visualization+and+localization+of+electrodes+used+in+electrophysiological+studies+of+brain+in+vivo.">Swept source optical coherence tomography as a tool for real time visualization and localization of electrodes used in electrophysiological studies of brain in vivo.</a> Biomedical Optics Express. 2, 3129-3134 (2011).</li><li>Huang, D., Swanson, E. A., Lin, C. P., Schuman, J. S., Stinson, W. G., Chang, W., Hee, M. R., Flottee, T., Gregory, K., Puliafito, C. A., Fujimoto, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+coherence+tomography.">Optical coherence tomography.</a> Science. 254, 1178-1181 (1991).</li><li>Mitsui, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+range+of+optical+reflectometry+with+spectral+interferometry.">Dynamic range of optical reflectometry with spectral interferometry.</a> Japanese Journal of Applied Physics. 38, 6133-6137 (1999).</li><li>de Boer, J. F., Cense, B., Park, B. H., Pierce, M. C., Tearney, G. J., Bouma, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+signal-to-noise+ratio+in+spectral-domain+compared+with+time-domain+optical+coherence+tomograhy.">Improved signal-to-noise ratio in spectral-domain compared with time-domain optical coherence tomograhy.</a> Optics Letters. 28, 2067-2069 (2003).</li><li>de Boer, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ch.+5.">Ch. 5.</a> Optical Coherence Tomography: Technology and Applications. , (2008).</li><li>Choma, M. A., Sarunic, M. V., Yang, C., Izatt, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensitivity+advantage+of+swept+source+and+fourier+domain+optical+coherence+tomography.">Sensitivity advantage of swept source and fourier domain optical coherence tomography.</a> Optics Express. 11, 2183-2189 (2003).</li><li>Leitgeb, R. A., Drexler, W., Unterhuber, A., Hermann, B., Bajraszewski, T., Le, T., Stingl, A., Fercher, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrahigh+resolution+fourier+domain+optical+coherence+tomography.">Ultrahigh resolution fourier domain optical coherence tomography.</a> Optics Express. 12, 2156-2165 (2004).</li><li>Drew, P. J., Shih, A. Y., Driscoll, J. D., Knutsen, P. M., Blinder, P., Davalos, D., Akassoglou, K., Tsai, P. S., Kleinfeld, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+optical+access+through+a+polished+and+reinforced+thinned+skull.">Chronic optical access through a polished and reinforced thinned skull.</a> Nature Methods. 7, 981-984 (2010).</li><li>Shih, A. Y., Mateo, C., Drew, P. J., Tsai, P. S., Kleinfeld, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Polished+and+Reinforced+Thinned-skull+Window+for+Long-term+Imaging+of+the+Mouse.">A Polished and Reinforced Thinned-skull Window for Long-term Imaging of the Mouse.</a> J. Vis. Exp. 61, e3742 (2012).</li><li>Yang, G., Pan, F., Parkhurst, C. N., Grutzendler, J., Gan, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thinned-skull+cranial+window+technique+for+long-term+imaging+of+the+cortex+in+live+mice.">Thinned-skull cranial window technique for long-term imaging of the cortex in live mice.</a> Nature Protocols. 5, (2010).</li><li>Lu, M., Majewska, S., K, A., Gelbard, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Thin-skull+Window+Technique+for+Chronic+Two-photon+In+vivo+Imaging+of+Murine+Microglia+in+Models+of+Neuroinflammation.">A Thin-skull Window Technique for Chronic Two-photon In vivo Imaging of Murine Microglia in Models of Neuroinflammation.</a> J. Vis. Exp. (43), e2059 (2010).</li><li>Wang, Y., Oh, C. M., Oliveira, M. C., Islam, M. S., Ortega, A., Park, B. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=GPU+accelerated+real-time+multi-functional+spectral-domain+optical+coherence+tomography+system+at+1300nm.">GPU accelerated real-time multi-functional spectral-domain optical coherence tomography system at 1300nm.</a> Optics Express. 20, 14797-14813 (2012).</li><li>Aguirre, A. D., Chen, Y., Fujimoto, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Depth-resolved+imaging+of+functional+activation+in+the+rat+cerebral+cortex+using+optical+coherence+tomography.">Depth-resolved imaging of functional activation in the rat cerebral cortex using optical coherence tomography.</a> Opt. Lett. 31, 3459-3461 (2006).</li><li>Chen, Y., Aguirre, A. D., Ruvinskaya, L., Devor, A., Boas, D. A., Fujimoto, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+coherence+tomography+(OCT)+reveals+depth-resolved+dynamics+during+functional+brain+activation.">Optical coherence tomography (OCT) reveals depth-resolved dynamics during functional brain activation.</a> Journal of Neuroscience Methods. 178, 162-173 (2009).</li><li>Liang, C., Wierwille, J., Moreira, T., Schwartzbauer, G., Jafri, M. S., Tang, C., Chen, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+forward-imaging+needle-type+OCT+probe+for+image+guided+stereotactic+procedures.">A forward-imaging needle-type OCT probe for image guided stereotactic procedures.</a> Opt Express. 19, 26283-26294 (2011).</li><li>Srinivasan, V. J., Sakadzic, S., Gorczynska, I., Ruvinskaya, S., Wu, W., Fugimoto, J. G., Boas, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+cerebral+blood+flow+with+optical+coherence+tomography.">Quantitative cerebral blood flow with optical coherence tomography.</a> Optics Express. 18, 2477-2494 (2010).</li><li>Galetta, K. M., Calabresi, P. A., Frohman, E. M., Balcer, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+Coherence+Tomography+(OCT):+imaging+the+visual+pathway+as+a+model+for+neurodegeneration.">Optical Coherence Tomography (OCT): imaging the visual pathway as a model for neurodegeneration.</a> The Journal of the American Society for Experimental NeuroTherapeutics. 8, 117-132 (2011).</li><li>Seigo, M. A., Sotirchos, E. S., Newsome, S., Babiarz, A., Eckstein, C., Ford, E., Oakley, J. D., Syc, S. B., Frohman, T. C., Ratchford, J. N., Balcer, L. J., Frohman, E. M., Calabresi, P. A., Saidha, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+assessment+of+retinal+neuronal+layers+in+multiple+sclerosis+with+maual+and+automated+optical+coherence+tomography+segementation+techniques.">In vivo assessment of retinal neuronal layers in multiple sclerosis with maual and automated optical coherence tomography segementation techniques.</a> J. Neurol. , (2012).</li><li>Frohman, E. M., Fujimoto, J. G., Frohman, T. C., Calabresi, P. A., Cutter, G., Balcer, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+coherence+tomography:+a+window+into+the+mechanisms+of+multiple+sclerosis.">Optical coherence tomography: a window into the mechanisms of multiple sclerosis.</a> Nature Clinical Practice. 4, 664-675 (2008).</li><li>Gill, A. S., Rajneesh, K. F., Owen, C. M., Yeh, J., Hsu, M., Binder, D. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+optical+detection+of+cerebral+edema+in+vivo.">Early optical detection of cerebral edema in vivo.</a> J. Neurosurg. 114, 470-477 (2011).</li></ol>

Imaging med OLT og en udtyndede kranium er en ny neuro-imaging teknik, der først lige er blevet undersøgt 15, 16. I vores eksperimenter viste vi muligheden for SD-oktober billeddannelse gennem en TSCW i en musemodel in vivo. Fra vores resultater, er kraniet fortyndes til cirka 55 um, og indtrængningsdybden opnås ved omtrent 1 mm med billedopløsning på 8 um og 20 um i aksial og lateral retning, hhv. I signalintensitet profil, øger oktober billeddannelse gennem en TSCW signalintensiteten og dybd...

Siden introduktionen i begyndelsen af 1990 er, er oktober vid udstrækning blevet anvendt til biologisk billeddannelse af væv struktur og funktion 2. Oktober genererer tværsnitsbilleder ved måling ekkotid forsinkelse af tilbagespredt lys 4 ved at gennemføre lave kohærens lyskilde med en fiberoptisk Michelson interferometer 2,4. SD-OLT, også kendt som Fourier-domæne OCT (FD-OLT), blev først introduceret i 1995 5 og tilbyder en overlegen afbildningsmodalitet sammenlignet med traditionel tid domæne OCT (TD-OLT). I SD-oktober, er referencearmen holdes stationær resulterer i en høj hastighed og ultrahøj opløsning billedoptagelse 6-9. I øjeblikket har TSCW modeller blevet anvendt i vid udstrækning til in vivo hjernescanning anvendelser af to-foton mikroskopi i stedet for en traditionel kraniotomi. Disse TSCW har været anvendt samtidig med en brugerdefineret dødningehoved plade eller et dækglas 10-13 at tilvejebringe yderligere imaging stabilitet. I vores undersøgelser har vi observeret, at tilbehør såsom disse ikke er nødvendige til OLT billeddannelse når en TSCW anvendes. Derfor er manglen på et kranium plade eller dækglas muliggør et bredere spektrum af billeddannende vinduesstørrelse, da de kan påvirke den optiske stråle og ændre oktober billeder. En tyndet-skull præparat har vist sig at være fordelagtig ved billeddannelse undersøgelser af hjernen ved hjælp af to-foton mikroskopi 10-13. I vores eksperimenter, benytter vi et SD-OLT-system til at afbilde cortex in vivo gennem en TSCW. Vores brugerdefinerede SD-oktober imaging setup indeholder en bredbåndsforbindelse, lav sammenhæng lyskilde bestående af to superluminescerende dioder (SLD) centreret ved 1295 nm med en båndbredde på 97 nm resulterer i en aksial og lateral opløsning på 8 um og 20 um, henholdsvis 14 . Med vores optisk billeddannelse enhed, forestiller vi, at billeddannelse gennem en TSCW har et stort potentiale i at identificere og visualisere strukturer og funktioner i optically tæt hjernevæv.

<table border="1"><tbody><tr><td> Materialer </td><td> Firma </td><td> Katalognummer </td><td> Kommentarer </td></tr><tr><td> Ketamin </td><td> Phoenix Pharmaceuticals </td><td> 57319-542-02 </td><td></td></tr><tr><td> Xylazin </td><td> Akorn, Inc. </td><td> 139-236 </td><td></td></tr><tr><td> Kunstige tårer Salve </td><td> Rugby </td><td> 0536-6550-91 </td><td></td></tr><tr><td> Nair </td><td> Church &amp; Dwight Co, Inc. </td><td> 4010130 </td><td></td></tr><tr><td> Sterile Alkohol Prep Pad </td><td> Kendall Healthcare </td><td> 6818 </td><td></td></tr><tr><td> Cotton tippes Applikatorer </td><td> Fisherbrand </td><td> 23-400-115 </td><td></td></tr><tr><td> Betadinopløsning vatpind </td><td> Purdue Produkter &lt;/ Td&gt; <td> 67618-153-01 </td><td></td></tr><tr><td> Saline Solution, .9% </td><td> Phoenix Pharmaceuticals </td><td> 57319-555-08 </td><td></td></tr><tr><td> Stereotaktisk Frame </td><td> Stoelting </td><td></td><td></td></tr><tr><td> High Speed ​​Kirurgisk håndboremaskine </td><td> Foredom </td><td></td><td> 38.000 rpm </td></tr><tr><td> Carbide Round Bur </td><td> Stoelting </td><td></td><td> 0,75 mm </td></tr><tr><td> Dura-grønne sten </td><td> Shofu </td><td></td><td> Shank: HP Form: BA1 </td></tr><tr><td> CompoMaster Grov &amp; CompoMaster Polermaskine </td><td> Shofu </td><td></td><td> Form: Mini-Pt. </td></tr><tr><td> SpaceDrapes </td><td> Braintree Scientific, Inc. </td><td></td><td></td></tr></tbody></table>

thinned-skull cortical window technique for in vivo optical coherence tomography imaging

Optisk kohærens tomografi (OCT) er en biomedicinsk billeddannelse teknik med høj rumlig-tidslig opløsning. Med sin minimalt invasiv tilgang oktober har været anvendt i udstrakt grad i oftalmologi, dermatologi, og gastroenterologi 1-3. Ved hjælp af en tyndere-skull kortikal vindue (TSCW) beskæftiger vi spektral-domæne OCT (SD-OLT) modalitet som et redskab til billede cortex in vivo. Almindeligvis har en åben-skull blevet anvendt til neuro-billeddannelse, da det giver større alsidighed dog en TSCW metode er mindre invasiv og et effektivt middel til langsigtet billeddannelse i neuropatologi undersøgelser. Her præsenteres en fremgangsmåde til at skabe en TSCW i en musemodel for in vivo oktober billeddannelse af hjernebarken.

Efter oprettelse af en fortyndet vindue over hjernebarken vaskulaturen bør nu være mere visuelt fremtrædende (fig. 1), og vil give mulighed for en dybere imaging dybde (op til 1 mm). Højre cortex fortyndes til cirka 55 um sammenlignet med en normal kranium målt ved 140 um (figur 1) og giver en større optisk klarhed. Yderligere udtynding til 10-15 um er mulig 11 dog ikke nødvendigt, da brug af dækglas og kranium plader er ikke implementeret i vores eksperimenter <stron...

Watch this Scientific Journal Video about Thinned-skull Cortical Window Technique for In Vivo Optical Coherence Tomography Imaging at JoVE.com

Thinned-skull Cortical Window Technique for In Vivo Optical Coherence Tomography Imaging

Vi præsenterer en fremgangsmåde til at skabe en tyndere-skull cortical vindue (TSCW) i en musemodel for<em&gt; In vivo</em&gt; Oktober billeddannelse af hjernebarken.

Tyndet-skull Cortical Window Teknik for<em&gt; In Vivo</em&gt; Optical Coherence Tomography Imaging

Optical coherence tomography (OCT) is a biomedical imaging technique  with high spatial-temporal resolution. With its minimally invasive approach OCT  has ...

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Neuroscience

Thinned-skull Cortical Window Technique for In Vivo Optical Coherence Tomography Imaging

A Thin-skull Window Technique for Chronic Two-photon In vivo Imaging of Murine Microglia in Models of Neuroinflammation

A Thin-skull Window Technique for Chronic Two-photon In vivo Imaging of Murine Microglia in Models of Neuroinflammation

Traditionally in neuroscience, in vivo two photon imaging of the murine central nervous system has either involved the use of open-skull1,2 or ...

Chronic Imaging of Mouse Visual Cortex Using a Thinned-skull Preparation

In vivo imaging using two-photon laser scanning microscopy (2PLSM) allows the study of living cells and neuronal processes in the intact brain. The ...

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In vivo imaging of cortical function requires optical access to the brain without disruption of the intracranial environment. We present a method to form ...

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In Vivo Two-photon Imaging Of Experience-dependent Molecular Changes In Cortical Neurons

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The brain's ability to change in response to experience is essential for healthy brain function, and abnormalities in this process contribute to a variety ...

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Two-Photon in vivo Imaging of Dendritic Spines in the Mouse Cortex Using a Thinned-skull Preparation

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In the mammalian cortex, neurons form extremely complicated networks and exchange information at synapses. Changes in synaptic strength, as well as ...

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Optical Coherence Tomography: Imaging Mouse Retinal Ganglion Cells In Vivo

Optisk kohærens tomografi: Imaging mus Retinal Ganglion celler In Vivo

Structural changes in the retina are common manifestations of ophthalmic diseases. Optical coherence tomography (OCT) enables their identification in ...

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In Vivo Two-photon Imaging of Cortical Neurons in Neonatal Mice

In Vivo To-foton billeddannelse af kortikale neuroner i Neonatal mus

Two-photon imaging is a powerful tool for the in vivo analysis of neuronal circuits in the mammalian brain. However, a limited number of in vivo imaging ...

Using Optical Coherence Tomography and Optokinetic Response As Structural and Functional Visual System Readouts in Mice and Rats

Ved hjælp af optisk kohærens tomografi og Optokinetic svar som strukturelle og funktionelle visuelle System udlæsninger i mus og rotter

Optical coherence tomography (OCT) is a fast, non-invasive, interferometric technique allowing high-resolution retinal imaging. It is an ideal tool for ...

Retinal Vascular Reactivity as Assessed by Optical Coherence Tomography Angiography

Retinal vaskulær reaktivitet som vurderet af optisk kohærens Tomografi Angiografi

The vascular supply to the retina has been shown to dynamically adapt through vasoconstriction and vasodilation to accommodate the metabolic demands of ...

Implantation of a Cranial Window for Repeated In Vivo Imaging in Awake Mice

Implantation of a Cranial Window for Repeated In Vivo Imaging in Awake Mice

Implantation af et kranievindue til gentagen In Vivo-billeddannelse i vågne mus

To fully understand the cellular physiology of neurons and glia in behaving animals, it is necessary to visualize their morphology and record their ...