Name: multimodal volumetric retinal imaging by oblique scanning laser ophthalmoscopy (oslo) and optical coherence tomography (oct)
Uploaded: 2018-08-04T14:30:00
Description: Watch this Scientific Journal Video about Multimodal Volumetric Retinal Imaging by Oblique Scanning Laser Ophthalmoscopy oSLO and Optical Coherence Tomography OCT at JoVE.com

Finansiering er fra Evans medicinsk fundament finansiering fra Boston Medical Center samt en sub-kontrakt fra NIH 5R01CA183101, BU-CTSI pilot give 1UL1TR001430, BU-Joslin pilotprogram og BU-CTSI KL2TR001411.

Alle metoder beskrevet her er blevet godkendt af Animal Care og brug udvalg (ACUC) af Boston Medical Center.
1. Systemopsætning
<ol><li>oSLO System<ol>
<li>Bruge en supercontinuum Laserkilde som system Laserkilde.<ol>
<li>Adskille det synlige lys spektrum (450-650 nm) fra højere bølgelængdeområdet (650-2000 nm) af en dichroic spejl (DM1). Udvide spektret med et par af dispersive prismer efter strålen passerer gennem en polarisering stråledeler (PBS).</li>
			<li>Sted en slids at vælg...

<ol>
	<li>Webb, R. H., Hughes, G. W., Delori, F. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Confocal+scanning+laser+ophthalmoscope.">Confocal scanning laser ophthalmoscope.</a> Applied Optics. 26 (8), 1492-1499 (1987).</li><li>Roorda, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adaptive+optics+scanning+laser+ophthalmoscopy.">Adaptive optics scanning laser ophthalmoscopy.</a> Optics Express. 10 (9), 405-412 (2002).</li><li>Huang, D., et al. <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 (5035), 1178-1181 (1991).</li><li>de Carlo, T. E., Romano, A., Waheed, N. K., Duker, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+optical+coherence+tomography+angiography+(OCTA).">A review of optical coherence tomography angiography (OCTA).</a> International Journal of Retina and Vitreous. 1 (1), 5 (2015).</li><li>Jia, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+Optical+Coherence+Tomography+Angiography+of+Choroidal+Neovascularization+in+Age-Related+Macular+Degeneration.">Quantitative Optical Coherence Tomography Angiography of Choroidal Neovascularization in Age-Related Macular Degeneration.</a> Ophthalmology. 121 (7), 1435-1444 (2014).</li><li>Chen, C. -. L., Wang, R. K. <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+based+angiography+[Invited].">Optical coherence tomography based angiography [Invited].</a> Biomedical Optics Express. 8 (2), 1056-1082 (2017).</li><li>Yi, J., Chen, S. Y., Shu, X., Fawzi, A. A., Zhang, H. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+retinal+imaging+using+visible-light+optical+coherence+tomography+guided+by+scanning+laser+ophthalmoscopy.">Human retinal imaging using visible-light optical coherence tomography guided by scanning laser ophthalmoscopy.</a> Biomedical Optics Express. 6 (10), 3701-3713 (2015).</li><li>Zhang, X. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dual-band+spectral-domain+optical+coherence+tomography+for+in+vivo+imaging+the+spectral+contrasts+of+the+retinal+nerve+fiber+layer.">Dual-band spectral-domain optical coherence tomography for in vivo imaging the spectral contrasts of the retinal nerve fiber layer.</a> Optics Express. 19 (20), 19653-19667 (2011).</li><li>Delori, F. C., Webb, R. H., Sliney, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maximum+permissible+exposures+for+ocular+safety+(ANSI+2000),+with+emphasis+on+ophthalmic+devices.">Maximum permissible exposures for ocular safety (ANSI 2000), with emphasis on ophthalmic devices.</a> Journal of the Optical Society of America a-Optics Image Science and Vision. 24 (5), 1250-1265 (2007).</li><li>Zhang, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Volumetric+fluorescence+retinal+imaging+in+vivo+over+a+30-degree+field+of+view+by+oblique+scanning+laser+ophthalmoscopy+(oSLO).">Volumetric fluorescence retinal imaging in vivo over a 30-degree field of view by oblique scanning laser ophthalmoscopy (oSLO).</a> Biomedical Optics Express. 9 (1), 25-40 (2018).</li></ol>

Her har vi beskrevet oSLO, en i vivo volumetriske fluorescerende retinal imaging teknik med en FOV over 30 °. I forhold til OLT, en aktuel standard for pleje imaging metode i oftalmologi, oSLO tilbyder en lignende 3D imaging kapacitet endnu tillader fluorescens kontrast, OCT ikke er følsomme over for. Fordelen ved oSLO er at det kræver kun en raster scanning, og herigennem tillader den sømløse kombination af OLT, give to supplerende teknikker til strukturelle og fluorescerende volumetriske billeddannelse.</...

Oftalmologi og vision videnskab stor gavn af de moderne optiske billeddannelse teknikker, da nethinden let kan tilgås med lys. Fluorescens nethinde billedbehandling er et vigtigt redskab i diagnosticeringen og forvaltning af chorioretinal Vaskulære sygdomme såsom diabetisk retinopati (DR) og aldersbetinget makuladegeneration (AMD), som begge er førende årsager til blindhed i USA.
Det er imidlertid stadig udfordrende at erhverve et stort synsfelt (FOV), tre-dimensionelle (3D) retinal imaging ved hjælp af fluorescens billeddannelse. Fundus fotografering har ikke mulighed for løsning af dybde og afviser ikke diffuse lys. Som et resultat, reducerer en blanding af signaler fra forskellige dybde billedkvaliteten. Scanning laser oftalmoskopi (SLO) og konfokal SLO (cSLO) kan reducere effekten af diffust lys ved hjælp af Konfokal gating1. Men det er vanskeligt for SLO eller cSLO at erhverve en 3D menneskelige retinal billede på grund af grænsen på deres dybden af fokus. Adaptive optik SLO (AOSLO) kan levere fremragende opløsning og kontrast ved at korrigere for wavefront aberrationer indført af det menneskelige øje. Imidlertid skulle AOSLO stadig z-stacking til volumetriske billeddannelse2. Optisk kohærens tomografi (OCT)3 og OCT angiografi (OCTA) systemer overvinde disse begrænsninger for at give tre-dimensionelle (3D) anatomiske og vaskulære billeder4,5,6, men naturens dye-fri OLT kan ikke visualisere lækage vejledende af vaskulær dysfunktion.
Denne protokol beskriver nye multimodale platform for 3D volumetriske fluorescens nethinde tænkelig, nemlig skråt scanning laser oftalmoskopi (oSLO). I denne billedbehandlingssystem, en skrå scanning genereres af en due hale skyderen, og en afsluttende billedbehandling system er justeret i en vinkel til at opdage fluorescens cross tværsnit billeder. Systemet bruger laser scanning metoder, og disse teknikker giver mulighed for nem integration med OCT som en supplerende volumetriske strukturelle imaging modalitet. Den nuværende dybde opløsning er ca. 25 µm i rotte nethinden og synsfeltet er 30°. Det væsentlige, oSLO giver mulighed for en fluorescerende version af OLT og samtidig kan kombineres med OCT og OCTA over en stor FOV.
I denne protokol, vil vi beskrive opsætningen af oSLO, metoden for tilpasning og konstruktion, metoden for i vivo billeddannelse af rotte nethinden og de repræsentative resultater.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Supercontinuum Laser Source</td>
			<td>NKT Photonics</td>
			<td>SuperK EXTREME EXU-OCT6</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichroic Mirror (DM1)</td>
			<td>Thorlabs</td>
			<td>DMLP650R</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichroic Mirror (DM2)</td>
			<td>Chroma</td>
			<td>ZT514/1064rpc</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichroic Mirror (DM3)</td>
			<td>Thorlabs</td>
			<td>DMLP900R</td>
			<td></td>
		</tr>
		<tr>
			<td>Single Mode Fiber (SMF 1)</td>
			<td>Thorlabs</td>
			<td>P3-460B-FC-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Single Mode Fiber (SMF 2)</td>
			<td>Thorlabs</td>
			<td>P3-780A-FC-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Optic Fiber Coupler</td>
			<td>Thorlabs</td>
			<td>TW850R5A2</td>
			<td></td>
		</tr>
		<tr>
			<td>1:1 Telescope System</td>
			<td>Thorlabs</td>
			<td>AC254-100-A&#215;2</td>
			<td></td>
		</tr>
		<tr>
			<td>3:1 Telescope System</td>
			<td>Thorlabs</td>
			<td>AC254-150-A&#215;2</td>
			<td></td>
		</tr>
		<tr>
			<td>3:1 Telescope System</td>
			<td>Thorlabs</td>
			<td>AC254-50-A&#215;2</td>
			<td></td>
		</tr>
		<tr>
			<td>1:1 Telescope System</td>
			<td>Thorlabs</td>
			<td>AC254-100-A&#215;2</td>
			<td></td>
		</tr>
		<tr>
			<td>Galvo Mirrors (GM1,GM2)</td>
			<td>Thorlabs</td>
			<td>GVS201&#215;2</td>
			<td></td>
		</tr>
		<tr>
			<td>De-sacn Galvo Mirrors (GM3)</td>
			<td>Thorlabs</td>
			<td>GVS011</td>
			<td></td>
		</tr>
		<tr>
			<td>Objective Lens</td>
			<td>Olympus</td>
			<td>UplanSApo 20&#215;/0.75</td>
			<td></td>
		</tr>
		<tr>
			<td>Final imaging system</td>
			<td>Olympus</td>
			<td>UplanFL N 10&#215;/0.3</td>
			<td></td>
		</tr>
		<tr>
			<td>Final imaging system</td>
			<td>Computar</td>
			<td>12-36mm/1:2.8</td>
			<td></td>
		</tr>
		<tr>
			<td>Camera</td>
			<td>PCO</td>
			<td>Pco.pixelfly usb</td>
			<td></td>
		</tr>
		<tr>
			<td>Filter</td>
			<td>Thorlabs</td>
			<td>FEL0800</td>
			<td></td>
		</tr>
		<tr>
			<td>Mounted Continuously Variable ND Filter</td>
			<td>Thorlabs</td>
			<td>NDC-50C-4M-A</td>
			<td></td>
		</tr>
		<tr>
			<td>Line Scan Camera</td>
			<td>Thorlabs</td>
			<td>SPL2048-140K</td>
			<td></td>
		</tr>
		<tr>
			<td>Analog Output Board (AO1)</td>
			<td>National Instrument</td>
			<td>PCI-6731</td>
			<td></td>
		</tr>
		<tr>
			<td>Analog Output Board (AO2)</td>
			<td>National Instrument</td>
			<td>PCIe-6351</td>
			<td></td>
		</tr>
		<tr>
			<td>Long pass filter</td>
			<td>Thorlabs</td>
			<td>FEL0800</td>
			<td></td>
		</tr>
	</tbody>
</table>

multimodal volumetric retinal imaging by oblique scanning laser ophthalmoscopy (oslo) and optical coherence tomography (oct)

Mens fluorescens imaging er meget udbredt i oftalmologi, er et stort synsfelt (FOV) tre-dimensionelle (3D) Fluorescens retinal billede stadig en stor udfordring med state-of-the-art retinal imaging modaliteter, fordi de ville kræve z-stacking til udarbejde en volumetrisk datasæt. Nyere optisk kohærens tomografi (OCT) og OCT angiografi (OCTA) systemer overvinde disse begrænsninger for at give tre-dimensionelle (3D) anatomiske og vaskulære billeder, men de dye-fri natur i OLT kan ikke visualisere lækage vejledende af vaskulære dysfunktion. Denne protokol beskriver en roman skrå scanning laser oftalmoskopi (oSLO) teknik, der giver 3D volumetriske fluorescens nethinde billedbehandling. Opsætningen af imaging systemet genererer den skrå scanning af en due hale skyderen og justerer den endelige billedbehandlingssystem i en vinkel til at opdage fluorescerende tværsnitsdata billeder. Systemet bruger laser scanning metode, og derfor tillader en nem indarbejdelse i OLT som en supplerende volumetriske strukturelle imaging modalitet. In vivo billeddannelse på rotte nethinden er vist her. Fluorescein løsning sprøjtes intravenøst for at producere volumetriske fluorescein angiografi (vFA).

Figur 4a viser en tværsnits OCT billede af en rotte nethinden. Figur 4b -4 c viser de samme retinal tværsnitsdata billeder af OCTA og oSLO vFA erhvervet på samme tid. OSLO muliggør tværsnits FA svarer til OCT B-scan. I forhold til OCTA identificerer oSLO vFA tværsnits billedet klart fartøjer i nerve fiber lag (NFL) og ganglion cellelag (GCL) og kapillærerne i yderste plexiform lag (OPL). <strong class="xf...

Watch this Scientific Journal Video about Multimodal Volumetric Retinal Imaging by Oblique Scanning Laser Ophthalmoscopy oSLO and Optical Coherence Tomography OCT at JoVE.com

Multimodal Volumetric Retinal Imaging by Oblique Scanning Laser Ophthalmoscopy oSLO and Optical Coherence Tomography OCT

Vi præsenterer her, en protokol for at få et stort synsfelt (FOV) tre-dimensionelle (3D) Fluorescens og OCT retinal billede ved hjælp af en roman imaging multimodal platform. Vi vil indføre Systemopsætning, metoden for tilpasning og de operationelle protokoller. In vivo billeddannelse vil blive demonstreret, og repræsentative resultater vil blive leveret.

Multimodal volumetriske Retinal Imaging af skrå Scanning Laser oftalmoskopi (oSLO) og optisk kohærens tomografi (OCT)

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Multimodal Volumetric Retinal Imaging by Oblique Scanning Laser Ophthalmoscopy (oSLO) and Optical Coherence Tomography (OCT)

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