Name: novel photoacoustic microscopy and optical coherence tomography dual-modality chorioretinal imaging in living rabbit eyes
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Dit werk werd gesteund door de genereuze steun van de 4K12EY022299 van het nationale Instituut van het oog (YMP), strijd voor zicht-International Retinal onderzoek Stichting FFS GIA16002 (YMP), onbeperkte departementale steun van onderzoek te voorkomen blindheid, en de Departement van de Universiteit van Michigan in oftalmologie en visuele wetenschappen. Dit werk gebruikt de Core Center voor Vision onderzoek gefinancierd door P30 EY007003 van het nationale Instituut van het oog.

Konijnen is dat een Amerikaanse ministerie van landbouw (USDA) bedekt soorten. Het gebruik ervan in biomedisch onderzoek moet aan strenge voorschriften. Alle konijn experimenten werden uitgevoerd overeenkomstig de verklaring van de ARVO (The Association for Research in visie en oogheelkunde) voor het gebruik van dieren in Ophthalmic en visie onderzoek, na goedkeuring van het laboratorium dierlijke protocol door de Universiteit Comité inzake gebruik en verzorging van dieren (UCUCA) van de Universiteit van Michigan (Proto...

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	<li>Wang, L. V., Hu, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photoacoustic+tomography:+in+vivo+imaging+from+organelles+to+organs.">Photoacoustic tomography: in vivo imaging from organelles to organs.</a> Science. 335 (6075), 1458-1462 (2012).</li><li>Beard, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomedical+photoacoustic+imaging.">Biomedical photoacoustic imaging.</a> Interface Focus. , (2011).</li><li>Taruttis, A., Ntziachristos, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+real-time+multispectral+optoacoustic+imaging+and+its+applications.">Advances in real-time multispectral optoacoustic imaging and its applications.</a> Nat Photonics. 9 (4), 219-227 (2015).</li><li>Tian, C., Xie, Z., Fabiilli, M. L., Wang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+and+sensing+based+on+dual-pulse+nonlinear+photoacoustic+contrast:+a+preliminary+study+on+fatty+liver.">Imaging and sensing based on dual-pulse nonlinear photoacoustic contrast: a preliminary study on fatty liver.</a> Opt Lett. 40 (10), 2253-2256 (2015).</li><li>Tian, C., 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-pulse+nonlinear+photoacoustic+technique:+a+practical+investigation.">Dual-pulse nonlinear photoacoustic technique: a practical investigation.</a> Biomed Opt Express. 6 (8), 2923-2933 (2015).</li><li>Tian, C., 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=Non-Contact+Photoacoustic+Imaging+Using+a+Commercial+Heterodyne+Interferometer.">Non-Contact Photoacoustic Imaging Using a Commercial Heterodyne Interferometer.</a> IEEE Sens J. 16 (23), 8381-8388 (2016).</li><li>Kim, K. 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Een intact en regelmatige traanfilm is essentieel voor kwalitatief hoogwaardige fundus beelden. Een onregelmatige en verslechterde scheur films kunnen aanzienlijk degraderen beeld kwaliteit42. Ter bescherming van de integriteit van de traanfilm en hoornvlies oppervlakkige punctate keratopathy voorkomen, is het essentieel om te smeren van het hoornvlies met boerenbedrog zeer vaak, ongeveer elke twee min. Als er bezorgdheid over de dekking van het oog, gebruik van een spleetlamp en fluoresceïne str...

Afgelopen decennia zijn getuige geweest van de explosieve ontwikkeling van het gebied van biomedische photoacoustic imaging1,2,3,4,5,6,7 ,8. Gebaseerd op de energieomzetting van licht in geluid, de opkomende beeldvorming van de photoacoustic kunt visualiseren biologische monsters op schalen van organellen, cellen, weefsels, organen voor het gehele lichaam kleine-dier en kan onthullen de anatomische functionele, moleculaire, genetische, en metabole informatie1,2,9,10,11,12. Photoacoustic imaging heeft unieke toepassingen gevonden in een waaier van biomedische velden, zoals cel biologie13,14, vasculaire biologie15,16, neurologie17,18 , oncologie19,20,21,22, Dermatologie23, farmacologie24en hematologie25,26. De toepassing ervan in de oogheelkunde, dat wil zeggen photoacoustic oogbeschadigingen en/of imaging, heeft aangetrokken aanzienlijke belangen van zowel wetenschappers en clinici en wordt momenteel actieve onderzocht.
In tegenstelling tot routinematig gebruikt oogbeschadigingen en/of imaging technologieën27, zoals fluoresceïne-angiografie (FA) en indocyanine groene angiografie (ICGA) (gebaseerd op fluorescentie contrast), optische coherentie tomografie (OCT) (gebaseerd op optische verstrooiing contrast) , en haar afgeleide OCT angiografie (gebaseerd op motie contrast van de rode bloedcellen), de photoacoustic oculair imaging gebruikt optische absorptie als het contrast-mechanisme. Dit is anders dan conventionele oogbeschadigingen en/of imaging-technologieën en biedt een uniek instrument voor de studie van optische absorptie-eigenschappen van het oog, die meestal geassocieerd met de pathofysiologische status van oogbeschadigingen en/of weefsel28 zijn. Tot op heden, aanzienlijke is uitstekend werk verricht in photoacoustic oogbeschadigingen en/of imaging29,30,31,32,33,34,35, 36,,37, maar deze studies richten op het achterste segment van de ogen van kleine dieren, zoals ratten en muizen. De baanbrekende studies tonen goed aan de haalbaarheid voor photoacoustic imaging in oogheelkunde, maar er is nog een lange weg te gaan naar klinische vertaling van de technologie sinds de oogbol maten van ratten en muizen zijn veel kleiner (minder dan eenderde) dan dat. van de mens. Als gevolg van de verspreiding van ultrasone golven over een aanzienlijk langere afstanden, kan signaal intensiteit en beeldkwaliteit sterk lijden wanneer de techniek wordt gebruikt voor de beeldvorming van het achterste segment van grotere ogen.
Richting van dit doel, we onlangs gemeld de noninvasive, etiket-vrije chorioretinal imaging in levende konijnen met behulp van geïntegreerde photoacoustic microscopie (PAM) en spectrale domein OCT (SD-OCT)38. Het systeem heeft uitstekende prestaties en het netvlies en vaatvlies van de ogen van grotere dieren op basis van endogene absorptie en verstrooiing contrast van oogbeschadigingen en/of weefsel kan visualiseren. Voorlopige resultaten bij konijnen laten zien dat de PAM noninvasively onderscheid kon maken tussen individuele netvlies en choroidal bloedvaten met behulp van een laser blootstelling dosis (~ 80 nJ) aanzienlijk lager ligt dan de veiligheidsgrens American National Standards Institute (ANSI) (160 nJ) bij 570 nm39; en de LGO kan duidelijk oplossen verschillende retinale lagen, het vaatvlies en de sclera. Het is de allereerste demonstratie van het achterste segment beeldvorming van grotere dieren met behulp van PAM en wellicht een belangrijke stap op weg naar klinische vertaling van de technologie, gezien het feit dat de grootte van de oogbol van konijnen (18,1 mm)40 bijna 80% van de axiale lengte van is mens (23.9 mm).
In dit werk, wij bieden een gedetailleerde beschrijving van de dual-modaliteit imaging systeem en experimentele protocollen die worden gebruikt voor noninvasive, etiket-vrije chorioretinal imaging in levende konijnen en tonen de systeemprestaties door representatieve Retina en choroidal imaging resultaten.

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	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Dual-modality imaging system</td>
			<td style="width:279px;"></td>
			<td style="width:157px;"></td>
			<td style="width:159px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">OPO laser</td>
			<td style="width:279px;">Ekspla (Vilnius, Lithuania)</td>
			<td style="width:157px;">NT-242</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Beam attenuator</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">AHWP10M-600</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Motorized rotation stage</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">PRM1/MZ8</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Motorized rotation stage controller</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">TDC001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Focusing lens</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">AC254-250-B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Pinhole</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">P50S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Collimating lens</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">AC127-030-B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Photodiode</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">PDA36A&#160;</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Laser shutter</td>
			<td style="width:279px;">Vincent Associates Inc. (Toronto, Canada)</td>
			<td style="width:157px;">LS6S2T0</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Laser shutter driver</td>
			<td style="width:279px;">Vincent Associates Inc. (Toronto, Canada)</td>
			<td style="width:157px;">VCM-D1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Dichroic mirror</td>
			<td style="width:279px;">Semrock, Inc. (Rochester, NY, USA)</td>
			<td style="width:157px;">Di03-R785-t3-25&#215;36</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Scan lens</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">OCT-LK3-BB</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Ophthalmic lens</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">AC080-010-B-ML</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Ultrasonic transducer</td>
			<td style="width:279px;">Optosonic Inc. (Arcadia, CA, USA)</td>
			<td style="width:157px;">Custom</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Amplifier</td>
			<td style="width:279px;">L3 Narda-MITEQ (Hauppauge, NY, USA)</td>
			<td style="width:157px;">AU-1647</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Band-pass filter</td>
			<td style="width:279px;">Mini-Circuits (Brooklyn, NY, USA)</td>
			<td style="width:157px;">BLP-30+</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Digitizer</td>
			<td style="width:279px;">DynamicSignals LLC (Lockport, IL, USA)</td>
			<td style="width:157px;">PX1500-4&#160;</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Synchronization electronics</td>
			<td style="width:279px;">National Instruments Corporation (Austin, TX, USA)</td>
			<td style="width:157px;">USB-6353</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">OCT module</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">Ganymede-II-HR</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Dispersion compensation glass</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">LSM03DC</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Illumination LED light</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">MCWHF2&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Power meter</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">S121C&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Power meter interface</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td>PM100USB&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Height measurement tool&#160;</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td>BHM1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Fundus camera</td>
			<td style="width:279px;">Topcon Corporation (Tokyo, Japan)&#160;</td>
			<td>TRC 50EX</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Matlab</td>
			<td style="width:279px;">MathWorks (Natick, MA, USA)</td>
			<td style="width:157px;">2017a</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Oscilloscope</td>
			<td style="width:279px;">Teledyne LeCroy (Chestnut Ridge, NY, USA)</td>
			<td style="width:157px;">WaveJet 354T</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Animal experiment</td>
			<td style="width:279px;"></td>
			<td style="width:157px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Water-circulating blanket</td>
			<td style="width:279px;">Stryker Corporation (Kalamazoo, MI, USA)</td>
			<td style="width:157px;">TP-700</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Ketamine hydrochloride injection</td>
			<td style="width:279px;">Par pharmaceutical, Inc. (Woodcliff Lake, NJ, USA)</td>
			<td style="width:157px;">NDC code 42023-115-10</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Xylazine hydrochloride</td>
			<td>VetOne (Boise, ID, USA)</td>
			<td style="width:157px;">NDC code 13985-704-10</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Tropicamide ophthalmic</td>
			<td style="width:279px;">Akorn Pharmaceuticals Inc. (Lake Forest, IL, USA)</td>
			<td style="width:157px;">NDC code 17478-102-12</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Phenylephrine hydrochloride ophthalmic</td>
			<td style="width:279px;">Paragon BioTeck, Inc. (Portland, OR, USA)</td>
			<td style="width:157px;">NDC code 42702-102-15</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Eye lubricant</td>
			<td style="width:279px;">Hub Pharmaceuticals LLC (Rancho Cucamonga, CA, USA)</td>
			<td style="width:157px;">NDC code 17238-610-15</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Eyewash</td>
			<td style="width:279px;">Altaire Pharmaceuticals, Inc. (Aquebogue, NY, USA)</td>
			<td style="width:157px;">NDC code 59390-175-18</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Tetracaine hydrochloride ophthalmic solution</td>
			<td style="width:279px;">Bausch &#38; Lomb, Inc. (Rochester, NY, USA)</td>
			<td style="width:157px;">NDC code 24208-920-64</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Flurbiprofen sodium ophthalmic solution</td>
			<td style="width:279px;">Bausch &#38; Lomb, Inc. (Rochester, NY, USA)</td>
			<td style="width:157px;">NDC code 24208-314-25</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:229px;">Neomycin and Polymyxin B Sulfates and Dexamethasone Ophthalmic Ointment</td>
			<td style="width:279px;">Bausch &#38; Lomb, Inc. (Rochester, NY, USA)</td>
			<td style="width:157px;">NDC code 24208-795-35</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Meloxicam injection</td>
			<td style="width:279px;">Henry Schein Inc. (Queens, NY, USA)</td>
			<td style="width:157px;">NDC code 11695-6925-1</td>
			<td></td>
		</tr>
	</tbody>
</table>

novel photoacoustic microscopy and optical coherence tomography dual-modality chorioretinal imaging in living rabbit eyes

Photoacoustic oogbeschadigingen en/of imaging is een opkomende ophthalmic imaging technologie die noninvasively oogbeschadigingen en/of weefsel door het licht energie omzetten in geluidsgolven kunt visualiseren en wordt momenteel intensief onderzocht. Echter de meeste gerapporteerd werk tot nu toe is gericht op de beeldvorming van het achterste segment van de ogen van kleine dieren, zoals ratten en muizen, die uitdagingen voor klinische menselijke vertaling als gevolg van kleine oogbol maten met zich meebrengt. Dit manuscript beschrijft een roman photoacoustic microscopie (PAM) en optische coherentie tomografie (OCT) dual-modaliteit systeem voor beeldvorming van het achterste segment van de ogen van grotere dieren, zoals konijnen. De configuratie van het systeem, systeem uitlijning, dierlijke voorbereiding en dual-modaliteit experimentele protocollen voor in vivo, noninvasive, etiket-vrije chorioretinal imaging bij konijnen zijn gedetailleerd. De doeltreffendheid van de methode wordt aangetoond door middel van representatieve experimentele resultaten, met inbegrip van therapieën van het netvlies en choroidal verkregen door de PAM en de LGO. Dit manuscript biedt een praktische gids voor de beeldvorming resultaten bij konijnen te reproduceren en bevordering van photoacoustic oogbeschadigingen en/of imaging in grotere dieren.

De dual-modaliteit imaging systeem en experimentele protocol zijn succesvol getest in de auteurs laboratorium met behulp van vier Nieuw-Zeeland wit konijnen. De volgende vitrines enkele representatieve resultaten.
Figuur 1 toont de schematische voorstelling van de PAM en SD-OCT dual-modaliteit imaging systeem. Het is samengesteld uit de volgende modules: photoacoustic licht bron, variabele laser ver...

Watch this Scientific Journal Video about Novel Photoacoustic Microscopy and Optical Coherence Tomography Dual-modality Chorioretinal Imaging in Living Rabbit Eyes at JoVE.com

Novel Photoacoustic Microscopy and Optical Coherence Tomography Dual-modality Chorioretinal Imaging in Living Rabbit Eyes

Dit manuscript beschrijft de nieuwe instelling en werkwijze van een photoacoustic microscopie en optische coherentie tomografie dual-modaliteit systeem voor noninvasive, etiket-vrije chorioretinal beeldvorming van grotere dieren, zoals konijnen.

Roman Photoacoustic microscopie en optische coherentie tomografie Dual-modaliteit Chorioretinal Imaging in levende konijn ogen

Photoacoustic ocular imaging is an emerging ophthalmic imaging technology that can noninvasively visualize ocular tissue by converting light energy into ...

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Engineering

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