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 arbeidet ble støttet av UC Discovery Proof of Concept stipend og ved NIH (R00 EB007241). Forfatterne vil også gjerne takke Jacqueline Hubbard for hennes assistanse i dette eksperimentet.

1. Kirurgisk Forberedelse <ol><li> Kvinne CD 1 mus i alderen 6-8 uker ble brukt i våre eksperimenter. </li><li> Anesthetize musen med en intraperitoneal injeksjon av en ketamin og xylazin kombinasjon (80 mg / kg ketamine/10 mg / kg xylazin). Plasser musen på en homeothermic pad for å sikre optimal kroppstemperatur på ~ 37 ° C. Kontinuerlig overvåker nivået av anestesi ved å teste dyr reflekser (f.eks, klemming fot med sløve tang) og injisere mer bedøvelse når det er nødvendig. </li><li> Smør begg...

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Bildebehandling med oktober og en tynnet-skull er en roman nevro-imaging teknikken som bare har nylig blitt undersøkt 15, 16. I våre eksperimenter, viste vi muligheten for SD-OCT avbildning gjennom en TSCW i en mus modell in vivo. Fra våre resultater, er skallen tynnet til ca 55 um og inntrengningsdybde oppnås ved ca 1 mm med bildeoppløsning av 8 um og 20 mikrometer i den aksiale og laterale retning, henholdsvis. I signalintensiteten profilen, øker oktober avbildning gjennom en TSCW signalinten...

Siden introduksjonen i begynnelsen av 1990-tallet har oktober vært mye brukt for biologisk avbildning av vev struktur og funksjon 2. Oktober genererer tverrsnitt bilder ved å måle ekko tidsforsinkelse av tilbakespredt lys 4 ved å implementere lav koherens lyskilde med en fiberoptisk Michelson interferometer 2,4. SD-OCT, også kjent som Fourier domene OCT (FD-OCT), ble først introdusert i 1995 5 og gir en overlegen imaging modalitet sammenliknet med tradisjonell tid domene OCT (TD-OCT). I SD-oktober er referansen armen holdes stasjonært resulterer i en høy hastighet og ultrahøy oppløsning bildeopptak 6-9. For tiden har TSCW modeller stor grad blitt brukt for in vivo avbildning av hjernen anvendelser av to-foton mikroskopi i stedet for et tradisjonelt kraniotomi. Disse TSCW har blitt brukt samtidig med en tilpasset hodeskalle plate eller et glass dekkglass 10-13 for å gi ekstra imaging stabilitet. I våre studier har vi sett at tilbehør som disse er ikke nødvendig for oktober bildebehandling når en TSCW brukes. Derfor tillater mangelen på en hodeskalle plate eller glass dekkglass for et bredere spekter av bildebehandling vinduet størrelse som de kan forstyrre den optiske strålen og endre oktober bilder. En tynnet-skull preparatet har vist seg å være fordelaktig i imaging studier av hjernen ved hjelp av to-foton mikroskopi 10-13. I våre eksperimenter, benytter vi et SD-OCT-systemet til bilde cortex in vivo gjennom en TSCW. Våre tilpassede SD-oktober bildebehandling oppsettet inneholder en bredbånd, lav koherens lyskilde bestående av to superluminescent dioder (SLD) sentrert ved 1295 nm med en båndbredde på 97 nm resulterte i en aksial og lateral oppløsning av 8 um og 20 um, henholdsvis 14 . Med vår optiske imaging-enhet, ser vi at avbildning gjennom en TSCW har et stort potensial i å identifisere og visualisere strukturer og funksjoner i optically tett hjernevev.

<table border="1"><tbody><tr><td> Materialer </td><td> Selskapet </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> Kunstig Tears 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> Steril alkohol Prep Pad </td><td> Kendall Healthcare </td><td> 6818 </td><td></td></tr><tr><td> Bomull Kommentar Applikatorer </td><td> Fisherbrand </td><td> 23-400-115 </td><td></td></tr><tr><td> Betadine Solution Swabstick </td><td> Purdue Produkter &lt;/ Td&gt; <td> 67618-153-01 </td><td></td></tr><tr><td> Saltvannsoppløsning, 0,9% </td><td> Phoenix Pharmaceuticals </td><td> 57319-555-08 </td><td></td></tr><tr><td> Stereotactic Frame </td><td> Stoelting </td><td></td><td></td></tr><tr><td> High Speed ​​Kirurgisk Hand Drill </td><td> Foredom </td><td></td><td> 38000 rpm </td></tr><tr><td> Carbide Round Bur </td><td> Stoelting </td><td></td><td> 0,75 mm </td></tr><tr><td> Dura-Green Stones </td><td> Shofu </td><td></td><td> Shank: HP Shape: BA1 </td></tr><tr><td> CompoMaster Grov &amp; CompoMaster polering </td><td> Shofu </td><td></td><td> Shape: 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 koherens tomografi (OCT) er en biomedisinsk bildebehandling teknikk med høy romlig-temporal oppløsning. Med sin minimalt invasiv tilnærming oktober har blitt brukt mye i oftalmologi, dermatologi, og gastroenterologi 1-3. Ved hjelp av en tynnet-skull kortikal vindu (TSCW), ansetter vi spektral-domene OCT (SD-OCT) modalitet som et verktøy for å avbilde cortex in vivo. Vanligvis har et åpnet-skull blitt brukt for nevro-avbildning som det gir mer allsidighet, er imidlertid en TSCW tilnærming mindre invasiv og er en effektiv gjennomsnittlig for langsiktig avbildning i neuropathology studier. Her presenterer vi en metode for å skape en TSCW i en mus modell for in vivo oktober avbildning av hjernebarken.

Etter oppretting av en tynnet vindu over hjernebarken blodkar skal nå være mer visuelt fremtredende (figur 1), og vil tillate en dypere avbildning dybde (opp til 1 mm). Høyre cortex er tynnet til ca 55 um i forhold til en normal hodeskalle målt ved 140 um (figur 1) og gir større optisk klarhet. Ytterligere tynning til 10-15 um er mulig 11 imidlertid ikke nødvendig da bruk av glass dekkglass og skallen plater ikke gjennomføres i våre eksperimenter (figur 1 og 2...

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 presenterer en metode for å skape en tynnet-skull kortikal vindu (TSCW) i en mus modell for<em&gt; In vivo</em&gt; Oktober avbildning av hjernebarken.

Tynnet-skull Cortical Window Teknikk for<em&gt; In Vivo</em&gt; Optisk koherens tomografi 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

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

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