Name: whole vitreous humor dissection for vitreodynamic analysis
Uploaded: 2015-05-24T14:00:00.000+00:00
Duration: 4 min 41 s
Description: Watch this Scientific Journal Video about Whole Vitreous Humor Dissection for Vitreodynamic Analysis at JoVE.com

The authors acknowledge the following funding sources, Whittier Foundation, Harrington Foundation, National Institutes of Health and Research to Prevent Blindness.

Tüm enükleasyon gözler bir mezbahaya elde edildi ve tüm deneyler kurumsal biyogüvenlik yasalarına uygun olarak yapıldı. <ol><li> Bir yüzey üzerinde enüklee gözü aşağı sabitleyin. Göz çevresinde aşırı doku yoluyla doku işaretçilerine yerleştirerek ve bir strafor tahta içine aşağı güvence bunu yapın. </li><li> Incelemek ve göz perilimbal konjonktivayı ayırın. <ol><li> Limbusta konjonktiva insizyon ve açık açık sklera kapalı incelemek için ince forseps (0.3 forseps) ve Microscissors (Westcott makas) kullanın. Künt diseksiyon daha diseksiyon izin ilerledikçe limbusun boyunca konjonktivayı kesin. </li><li> Mümkün olduğunca sklera açığa için her yolu göze (360 derece) etrafında konjonktivayı çıkarın. NOT: Bu prosedür geri kalanı boyunca, cerrahi iğne veya bir forseps ile göz fikse kolaylaştırmak için konjonktiva küçük bir miktar terk etmek yararlı olur. </li></ol></li><li> Biri boyunca tam kalınlıkta skleral flep oluşturmagöz (neşter bıçağı 69) yan. <ol><li> Yavaşça koyu pigmentli koroid kadar bir neşter (neşter bıçağı 11) ile sklera içine keserek limbusa limbusun ve 3 mm posterior bir ~ 5 mm skleral kesi paralel hale görünür. Koroid kendisi kesilirken için dikkatli olun. </li><li> Yavaşça içeri koroid iterek bu dokular arasındaki potansiyel boşluk büyütmek için mümkün kadar dikkatlice, (künt diseksiyon kullanarak) makasla veya neşter ile ya sklera ve koroid arasındaki düzlemi boyunca teşrih. </li><li> T-şekilli kesi oluşturarak, keskin microscissors ilk skleral kesi dik bir skleral kesi yapmak. </li><li> Sonra açık açık yatan koroid gelen skleral doku kaldırmak ve yavaşça yukarıda belirtildiği gibi sklera uzak koroid itmek için bir çevresel biçimde incelemek için devam ediyor. </li><li> Gerektiği gibi skleral flep büyütün. <ol><li> Yavaşça uzak skleradan koroid teşrih küntsüreci boyunca. </li><li> Limbusun dik yapılan kesi optik sinir ulaşır ve kesi limbusa paralel göz çevresi (45 °) en az üçte biri kadar yapılan flep büyütün. </li><li> Künt diseksiyon sırasında göz işlemek için kaldıraç kaynağı olarak skleral flep kullanın. kapağın karşı çekiş künt diseksiyon için kolaylaştırır. </li></ol></li><li> Diğer skleral flep önceki adımı yineleyin. </li></ol></li><li> Koroid geniş bir alana maruz skleral flep kesip. </li><li> Gözün çevresi (360 derece) etrafında 3. adımda yapılan kesi devam edin. </li><li> Kalan koroid-retina doku çıkarın. <ol><li> Maruz Bölgede, hafifçe geriye kalan koroid-retina dokusu savmak için bir pamuk-ucu kullanın. Alternatif olarak, hafifçe forseps ile doku kapmak ve altta yatan vitreus kapalı soyulması. </li><li> Gerekirse koroid st içine bir kesi yapmakoptik sinir Arting. Sonra retina ve vitreus korteksi maruz yavaşça koroid uzak soyun. </li></ol></li><li> Uzakta skleradan koroid diseksiyon künt Devam ve daha sonra tüm bozulmadan vitrözü elde etmek koroid uzak soyun. </li><li> İstenilen yerde bütün vitrözü konumlandırmak için korneaya bağlı skleral jant kullanın. Bu noktada, vitröz ön sklera ve mercek takılır. </li><li> Tüm sklera kaldırarak, lens etrafında, sklera içinden vitrözü teşrih künt. </li><li> Gerekirse uzak vitreus lensi kepçe künt bir alet kullanın. Çeşitli vitreodynamic deneyler için örnek kullanın. <ol><li> Havaya maruz kalan yüzey alanı, bilinen bir cam beher içinde örnek yerleştirin. Numunenin kenarında oksijen duyarlı probu yerleştirin. Örnek olarak bilinen bir mesafe (R) probu hareket ettirmek için, bir mikro-manipülatör kullanın. </li><li> Teorik difüzyon denklemini elde etmek için difüzyon Fick yasaları kullanın. Dekar ileta adımda 10.1 toplanan vb deneysel difüzyon katsayısı / reaksiyon süresini elde </li></ol></li></ol>

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The authors have nothing to disclose.

Dikkatle vitreus diseksiyonu sırasında yapılmalıdır iki kritik adımlar vardır. Tam kalınlıkta skleral flep oluşturma Adım 3, tüm diseksiyon için çok önemlidir. Bakım tam kalınlıkta skleral kanatçık oluştururken koroid halinde kesmek için önlem alınmalıdır. diğer kritik adım koroid gelen sklera uzak diseksiyon olduğunu. Bu adım dikkatle vitreus dışarı dökmek olabilir hangi koroid birden delikler oluşturarak önlemek için yapılmalıdır. Protokolü değiştirmek ve hala bozulmamış t?...

Bu yöntemin amacı, detay vitreodynamic analiz amaçları için, kadavra göz, camsı çekirdek ve korteks bozulmamış bir bütün dokunulmamış vitreus izole etmek için kullanılan bir tekniktir. Vitreus fizyoloji alanında böyle akışkanlar mekaniği araştırmacılar olarak, çok disiplinli araştırmacı, büyüdü gibi, vitreus 1 fiziksel ve biyomekanik özelliklerini araştırıyor. Bu amaçla, detaylara multi-disipliner araştırmacılar yardımcı olmak için bütün, sağlam camsı vücudu izole etmek için bir tekniktir esastır. Sebag ve ark., 2. ve diğerleri 3 insan kadavra göze şık bütün vitreus diseksiyonu yapılan ve sonuçları illüstrasyonlar gösterdi. Ancak, kullanılan teknik, bağımsız yöntemi çoğaltmak mümkün olmaz detay ve uzman olmayanların tarif edilmemiştir. Diğer çalışmalar, böyle bir aspirasyon veya kısmi diseksiyonu gibi basit yöntemler kullanılarak kadavra gözlerinden vitreus hasat varher ikisi de bütün dokunulmamış vitreus neden yoktur. Gisladottis ve ark., 4 ve Xu ve ark. 5 kadavra gözlerinden hasat vitreus içinde geçirgenliği araştırmak. Camsı ekstraksiyon hiç bir yöntem tarif edilmiştir Bununla birlikte, bu bir şırınga ile vitreus aspire varsayılmıştır. Watts ve ark., 6 cerrahi teknikle tavşan vitreus mizah izole bir yöntem anlatarak bir adım daha ileri gitti. Bununla birlikte, bu yöntem sadece camsı çekirdek bir yalıtım olup camsı korteks ile sonuçlanır. Skeie ve ark., 7 sonra 4 eşsiz bölgelere vitrözü organize ve zarif analiz için her bölümünü incelemek için bir yöntem tarif. Bu teknik, bununla birlikte, bir bütün olarak, bozulmamış bir vitröz ile sonuçlanmaz. Geçerli bir teknik şu anda sadece kadavra gözlerinde yapılır biyofizik deneyler kolaylaştırmak için geliştirilmiştir. Önceki yöntem olarak tarif edilen birBove, 1) hiçbiri tamamen bütün vitreus vücudu izole çünkü sınırlıdır, 2) hasat vitreus çekirdek ve korteks 3) vitreus anatomik yapısı korunur değil, homojenize, ya da 4) diseksiyon teknikleri yeterince diğer alanlardaki araştırmacılar tarafından çoğaltma için ayrıntılı değil . Buna ek olarak, sklera ve koroid, vitreus görselleştirme opaklığına nedeniyle sağlam gözün sınırlıdır. Bütün bu gözün içine yapılabilir ölçümlerin hassasiyetini ve fizibilite sınırlar. Buna ek olarak, vitrözü çevreleyen anatomik yapılar vitreus biyokimyasal ve fiziksel özelliklerinin çalışma karıştırabilir. Son yıllarda, camsı bilimin vücut müthiş büyüdü ve tüm vitreus bireysel parçaları farklı özelliklere sahip olduğuna inanmak için bir neden yoktur. Vitreodynamics Araştırma, için vitreus fiziksel biyomekanik ve kimyasal özellikleri araştıran artan bir ilgi vardırBöyle ilaç dağıtım, intravitreal oksijenlenme 8 ve vitrektomi gibi klinik tıpta uygulamaları vardır h. Vitrözü işlemek için farmakolojik ajanlar kullanan Farmakolojik vitreodynamics, vitrektomi sonuçları 9 iyileştirmek için kullanılabilir. Biyomekanik özellikleri intravitreal ilaç verme teknolojileri 10-12 geliştirmek için kullanılabilir camsı sıvı akışı, modellemek için kullanılır. Vitreus çeşitli kesimleri fiziksel özellikleri vitro-retinal oksijen taşıma 13 anlaşılması açısından büyük önem taşıyor. Önerilen vitreus diseksiyon tekniği bozulmadan vitreus çeşitli özelliklerini incelemek için kullanılabilir. Bu tezgah üstü deneyleri daha iyi görselleştirme ile tüm, sağlam camsı organları yapılması mümkün kılar. Özet olarak, vitröz çalışma için mevcut yöntemler ya yeterince tarif değildir, ya da eksik bir izolasyon camsı nüve ve korteks ile sonuçlanır. Bu nedenle, e gerçekleştirmek için bir ihtiyaç vardırşeffaf göz modelinde xperiments kadavra gözün var vitreus anatomisi korurken.

<table><tbody><tr><td>0.3 forceps</td><td>Storz Opthalmics</td><td>E1793</td><td></td></tr><tr><td>Westcott Tenotomy Scissors Curved Right</td><td>Storz Opthalmics</td><td>E3320 R</td><td></td></tr><tr><td>Scalpel Handle No. 3</td><td>VWR</td><td>25607-947</td><td></td></tr><tr><td>Scalpel Blade, #11, for #3 Handle</td><td>VWR</td><td>470174-844</td><td></td></tr></tbody></table>

whole vitreous humor dissection for vitreodynamic analysis

The authors propose an effective technique to isolate whole, intact vitreous core and cortex from post mortem enucleated porcine eyes. While previous studies have shown the results of such dissections, the detailed steps have not been described, precluding researchers outside the field from replicating their methods. Other studies harvest vitreous either through aspiration, which does not maintain the vitreous structure anatomy, or through partial dissection, which only isolates the vitreous core. The proposed method isolates the whole vitreous body, with the vitreous core and cortex intact, while maintaining vitreous anatomy and structural integrity. In this method, a full thickness scleral flap in an enucleated porcine eye is first created and through this, the choroid tissue can be separated from the sclera. The scleral flap is then expanded and the choroid is completely separated from the sclera. Finally the choroid-retina tissue is peeled off the vitreous to leave an isolated intact vitreous body. The proposed vitreous dissection technique can be used to study physical properties of the vitreous humor. In particular, this method has significance for experimental studies involving drug delivery, vitreo-retinal oxygen transport, and intraocular convection.

Protokol sonrasında sağlam çekirdek ve korteks (Şekil 3) ile başarılı bir vitreus diseksiyon yol açacaktır. Bu vitröz korteks yapıştırılmış retina kalıntı parçalar bellidir. Bozulmamış tam vitröz hümör, belirli vitreodynamic deneyler için çeşitli şekillerde kullanılabilir. Bizim durumumuzda, difüzyon sağlam vitreus oksijen oranı ve karşılık gelen zaman sabiti (Şekil 2) incelendi var. Bizim yöntemi kullanılarak (çekirdek ve korteks) ayrılmıştır ...

Watch this Scientific Journal Video about Whole Vitreous Humor Dissection for Vitreodynamic Analysis at JoVE.com

Whole Vitreous Humor Dissection for Vitreodynamic Analysis

The goal of this protocol is to show an effective technique to isolate whole, intact vitreous core and cortex from post mortem enucleated porcine eyes.

Vitreodynamic Analiz Tüm Vitreus Mizah Diseksiyon

The authors propose an effective technique to isolate whole, intact vitreous core and cortex from post mortem enucleated porcine eyes. While previous ...

whole-vitreous-humor-dissection-for-vitreodynamic-analysis

Research

JoVE Journal

Medicine

11.3K Görüntüleme.  University of Southern California. The goal of this protocol is to show an effective technique to isolate whole, intact vitreous core and cortex from post mortem enucleated porcine eyes.

Video: Whole Vitreous Humor Dissection for Vitreodynamic Analysis

Dissection of Human Vitreous Body Elements for Proteomic Analysis

The vitreous is an optically clear, collagenous extracellular matrix that fills the inside of the eye and overlies the retina. 1,2 Abnormal interactions between vitreous substructures and the retina underlie several vitreoretinal diseases, including retinal tear and detachment, macular pucker, macular hole, age-related macular degeneration, vitreomacular traction, proliferative vitreoretinopathy, proliferative diabetic retinopathy, and inherited vitreoretinopathies. 1,2 The molecular composition of the vitreous substructures is not known. Since the vitreous body is transparent with limited surgical access, it has been difficult to study its substructures at the molecular level. We developed a method to separate and preserve these tissues for proteomic and biochemical analysis. The dissection technique in this experimental video shows how to isolate vitreous base, anterior hyaloid, vitreous core, and vitreous cortex from postmortem human eyes. One-dimensional SDS-PAGE analyses of each vitreous component showed that our dissection technique resulted in four unique protein profiles corresponding to each substructure of the human vitreous body. Identification of differentially compartmentalized proteins will reveal candidate molecules underlying various vitreoretinal diseases.

This video shows an effective technique for differentiating and dissecting the various semi-transparent structures of the human vitreous body in post mortem eyes.

Proteomik Analizi İnsan Vitreus Gövde Elemanları diseksiyonu

The vitreous is an optically clear, collagenous extracellular matrix that fills the inside of the eye and overlies the retina. 1,2 Abnormal interactions ...

Tıp

Hücresiz DNA Kullanılarak Vitreoretinal Lenfoma Tanısı

Cell-Free DNA Extraction of Vitreous and Aqueous Humor Specimens for Diagnosis and Monitoring of Vitreoretinal Lymphoma

Vitreoretinal lymphoma (VRL) represents an aggressive lymphoma, often categorized as primary central nervous system diffuse large B-cell lymphoma. To diagnose VRL, specimens such as vitreous humor and, more recently, aqueous humor are collected. Diagnostic testing for VRL on these specimens includes cytology, flow cytometry, and molecular testing. However, both cytopathology and flow cytometry, along with molecular testing using cellular DNA, necessitate intact whole cells. The challenge lies in the fact that vitreous and aqueous humor typically have low cellularity, and many cells get destroyed during collection, storage, and processing. Moreover, these specimens pose additional difficulties for molecular testing due to the high viscosity of vitreous humor and the low volume of both vitreous and aqueous humor. This study proposes a method for extracting cell-free DNA from vitreous and aqueous specimens. This approach complements the extraction of cellular DNA or allows the cellular component of these specimens to be utilized for other diagnostic methods, including cytology and flow cytometry.

Yazar Spotlight: Vitreus Mizahından Hücresiz DNA Ekstraksiyonu Kullanarak VRL Teşhisini İlerletme 

A procedure to extract cell-free DNA from vitreous and aqueous humor to perform molecular studies for diagnosing vitreoretinal lymphoma is established here. The method offers the ability to concurrently extract DNA from the cellular component of the sample or to reserve it for ancillary testing.

Vitreoretinal Lenfoma Tanı ve İzlemi için Vitreus ve Sulu Mizah Örneklerinin Hücresiz DNA Ekstraksiyonu

Vitreoretinal lymphoma (VRL) represents an aggressive lymphoma, often categorized as primary central nervous system diffuse large B-cell lymphoma. To ...

Kanser Araştırmaları

Evisceration of Mouse Vitreous and Retina for Proteomic Analyses

While the mouse retina has emerged as an important genetic model for inherited retinal disease, the mouse vitreous remains to be explored. The vitreous is a highly aqueous extracellular matrix overlying the retina where intraocular as well as extraocular proteins accumulate during disease.1-3 Abnormal interactions between vitreous and retina underlie several diseases such as retinal detachment, proliferative diabetic retinopathy, uveitis, and proliferative vitreoretinopathy.1,4 The relative mouse vitreous volume is significantly smaller than the human vitreous (Figure 1), since the mouse lens occupies nearly 75% of its eye.5 This has made biochemical studies of mouse vitreous challenging. In this video article, we present a technique to dissect and isolate the mouse vitreous from the retina, which will allow use of transgenic mouse models to more clearly define the role of this extracellular matrix in the development of vitreoretinal diseases.

The dissection technique illustrates evisceration of the vitreous, retina, and lens from the mouse eye, separation by centrifugation, and characterization with protein assays.

Proteomik Analizleri Fare Vitreus ve Retina eviserasyon

While the mouse retina has emerged as an important genetic model for inherited retinal disease, the mouse vitreous remains to be explored. The vitreous is ...

Biyoloji

A Step by Step Protocol for Subretinal Surgery in Rabbits

Age related macular degeneration (AMD), retinitis pigmentosa, and other RPE related diseases are the most common causes for irreversible loss of vision in adults in industrially developed countries. RPE transplantation appears to be a promising therapy, as it may replace dysfunctional RPE, restore its function, and thereby vision.
Here we describe a method for transplanting a cultured RPE monolayer on a scaffold into the subretinal space (SRS) of rabbits. After vitrectomy xenotransplants were delivered into the SRS using a custom made shooter consisting of a 20-gauge metallic nozzle with a polytetrafluoroethylene (PTFE) coated plunger. The current technique evolved in over 150 rabbit surgeries over 6 years. Post-operative follow-up can be obtained using non-invasive and repetitive in vivo imaging such as spectral domain optical coherence tomography (SD-OCT) followed by perfusion-fixed histology.
The method has well-defined steps for easy learning and high success rate. Rabbits are considered a large eye animal model useful in preclinical studies for clinical translation. In this context rabbits are a cost-efficient and perhaps convenient alternative to other large eye animal models.

Retinal pigment epithelium (RPE) replacement strategies and gene-based therapy are considered for several retinal degenerative conditions. For clinical translation, large eye animal models are required to study surgical techniques applicable in patients. Here we present a rabbit model for subretinal surgery geared towards RPE transplantation, which is versatile and cost-efficient.

Tavşanlarda Subretinal Cerrahi Adım Protokolle Bir Adım

Age related macular degeneration (AMD), retinitis pigmentosa, and other RPE related diseases are the most common causes for irreversible loss of vision in ...

Microsurgical Dissection and Tissue Clearing for High Resolution Intact Whole Retina and Vitreous Imaging

Neuronal and vascular structures of the retina in physiologic and pathologic conditions can be better visualized and characterized by using intact whole retina imaging techniques compared to conventional retinal flat mount preparations and sections. However, immunofluorescent imaging of intact whole retina is hindered by the opaque coatings of the eyeball, i.e., sclera, choroid, and retinal pigment epithelium (RPE) and the light scattering properties of retinal layers that prevent full thickness high resolution optical imaging. Chemical bleaching of the pigmented layers and tissue clearing protocols have been described to address these obstacles; however, currently described methods are not suitable for imaging endogenous fluorescent molecules such as green fluorescent protein (GFP) in intact whole retina. Other approaches bypassed this limitation by surgical removal of pigmented layers and the anterior segment of the eyeball allowing intact eye imaging, though the peripheral retina and hyaloid structures were disrupted. Presented here is an intact whole retina and vitreous immunofluorescent imaging protocol that combines surgical dissection of the sclera/choroid/retina pigment epithelium (RPE) layers with a modified tissue clearing method and light sheet fluorescent microscopy (LSFM). The new approach offers an unprecedented view of&#160;unperturbed vascular and neuronal elements of the retina as well as the vitreous and hyaloid vascular system in pathologic conditions.

Presented here is a protocol for intact whole retina imaging in which the outer opaque/pigmented layers of the eyeball are surgically removed, and optical clearing is applied to render retina transparent enabling the visualization of the peripheral retina and hyaloid vasculature in intact retina using light sheet fluorescent microscopy.

Neuronal and vascular structures of the retina in physiologic and pathologic conditions can be better visualized and characterized by using intact whole ...

Nörobilim

Full-Circle Cauterization of Limbal Vascular Plexus for Surgically Induced Glaucoma in Rodents

Glaucoma, the second leading cause of blindness worldwide, is a heterogeneous group of ocular disorders characterized by structural damage to the optic nerve and retinal ganglion cell (RGC) degeneration, resulting in visual dysfunction by interrupting the transmission of visual information from the eye to the brain. Elevated intraocular pressure is the most important risk factor; thus, several models of ocular hypertension have been developed in rodents by either genetic or experimental approaches to investigate the causes and effects of the disease. Among those, some limitations have been reported such as surgical invasiveness, inadequate functional assessment, requirement of extensive training, and highly variable extension of retinal damage. The present work characterizes a simple, low-cost, and efficient method to induce ocular hypertension in rodents, based on low-temperature, full-circle cauterization of the limbal vascular plexus, a major component of aqueous humor drainage. The new model provides a technically easy, noninvasive, and reproducible subacute ocular hypertension, associated with progressive RGC and optic nerve degeneration, and a unique post-operative clinical recovery rate that allows in vivo functional studies by both electrophysiological and behavioral methods.

The goal of this protocol is to characterize a novel model of glaucomatous neurodegeneration based on 360&#176; thermic cauterization of limbal vascular plexus, inducing subacute ocular hypertension.

Kemirgenlerde Cerrahi Olarak İndüklenen Glokom için Limbal Vasküler Pleksusun Tam Daire Koterizasyonu

Glaucoma, the second leading cause of blindness worldwide, is a heterogeneous group of ocular disorders characterized by structural damage to the optic ...

Retinal Pigment Epithelium Transplantation in a Non-human Primate Model for Degenerative Retinal Diseases

Retinal pigment epithelial (RPE) transplantation holds great promise for the treatment of inherited and acquired retinal degenerative diseases. These conditions include retinitis pigmentosa (RP) and advanced forms of age-related macular degeneration (AMD), such as geographic atrophy (GA). Together, these disorders represent a significant proportion of currently untreatable blindness globally. These unmet medical needs have generated heightened academic interest in developing methods of RPE replacement. Among the animal models commonly utilized for preclinical testing of therapeutics, the non-human primate (NHP) is the only animal model that has a macula. As it shares this anatomical similarity with the human eye, the NHP eye is an important and appropriate preclinical animal model for the development of advanced therapy medicinal products (ATMPs) such as RPE cell therapy.
This manuscript describes a method for the submacular transplantation of an RPE monolayer, cultured on a polyethylene terephthalate (PET) cell carrier, underneath the macula onto a surgically created RPE wound in immunosuppressed NHPs. The fovea-the central avascular portion of the macula-is the site of the greatest mechanical weakness during the transplantation. Foveal trauma will occur if the initial subretinal fluid injection generates an excessive force on the retina. Hence, slow injection under perfluorocarbon liquid (PFCL) vitreous tamponade is recommended with a dual-bore subretinal injection cannula at low intraocular pressure (IOP) settings to create a retinal bleb. 
Pretreatment with an intravitreal plasminogen injection to release parafoveal RPE-photoreceptor adhesions is also advised. These combined strategies can reduce the likelihood of foveal tears when compared to conventional techniques. The NHP is a key animal model in the preclinical phase of RPE cell therapy development. This protocol addresses the technical challenges associated with the delivery of RPE cellular therapy in the NHP eye.

The non-human primate (NHP) is an ideal model for studying human retinal cellular therapeutics due to the anatomical and genetic similarities. This manuscript describes a method for submacular transplantation of retinal pigment epithelial cells in the NHP eye and strategies to prevent intraoperative complications associated with macular manipulation.

Dejeneratif Retina Hastalıkları için İnsan Dışı Primat Modelinde Retina Pigment Epitel Transplantasyonu

Retinal pigment epithelial (RPE) transplantation holds great promise for the treatment of inherited and acquired retinal degenerative diseases. These ...

AMD Araştırması için İnsan Donör Gözlerinin OCT Tabanlı Multimodal Görüntülemesi

Ex Vivo OCT-Based Multimodal Imaging of Human Donor Eyes for Research into Age-Related Macular Degeneration

A progression sequence for age-related macular degeneration (AMD) learned from optical coherence tomography (OCT)-based multimodal (MMI) clinical imaging could add prognostic value to laboratory findings. In this work, ex vivo OCT and MMI were applied to human donor eyes prior to retinal tissue sectioning. The eyes were recovered from non-diabetic white donors aged &#8805;80 years old, with a death-to-preservation time (DtoP) of &#8804;6 h. The globes were recovered on-site, scored with an 18 mm trephine to facilitate cornea removal, and immersed in buffered 4% paraformaldehyde. Color fundus images were acquired after anterior segment removal with a dissecting scope and an SLR camera using trans-, epi-, and flash illumination at three magnifications. The globes were placed in a buffer within a custom-designed chamber with a 60 diopter lens. They were imaged with spectral domain OCT (30&#176; macula cube, 30 &#181;m spacing, averaging = 25), near-infrared reflectance, 488 nm autofluorescence, and 787 nm autofluorescence. The AMD eyes showed a change in the retinal pigment epithelium (RPE), with drusen or subretinal drusenoid deposits (SDDs), with or without neovascularization, and without evidence of other causes. Between June 2016 and September 2017, 94 right eyes and 90 left eyes were recovered (DtoP: 3.9 &plusmn; 1.0 h). Of the 184 eyes, 40.2% had AMD, including early intermediate (22.8%), atrophic (7.6%), and neovascular (9.8%) AMD, and 39.7% had unremarkable maculas. Drusen, SDDs, hyper-reflective foci, atrophy, and fibrovascular scars were identified using OCT. Artifacts included tissue opacification, detachments (bacillary, retinal, RPE, choroidal), foveal cystic change, an undulating RPE, and mechanical damage. To guide the cryo-sectioning, OCT volumes were used to find the fovea and optic nerve head landmarks and specific pathologies. The ex vivo volumes were registered with the in vivo volumes by selecting the reference function for eye tracking. The ex vivo visibility of the pathology seen in vivo depends on the preservation quality. Within 16 months, 75 rapid DtoP donor eyes at all stages of AMD were recovered and staged using clinical MMI methods.

Yazar Spotlight: Yaşa Bağlı Makula Dejenerasyonu Araştırmaları için İnsan Donör Gözlerinin Ex Vivo OCT Tabanlı Multimodal Görüntülemesi  

Laboratory assays can leverage prognostic value from the longitudinal optical coherence tomography (OCT)-based multimodal imaging of age-related macular degeneration (AMD). Human donor eyes with and without AMD are imaged using OCT, color, near-infrared reflectance scanning laser ophthalmoscopy, and autofluorescence at two excitation wavelengths prior to tissue sectioning.

Ex Vivo Yaşa Bağlı Makula Dejenerasyonu Araştırmaları için İnsan Donör Gözlerinin OCT Tabanlı Multimodal Görüntülemesi

A progression sequence for age-related macular degeneration (AMD) learned from optical coherence tomography (OCT)-based multimodal (MMI) clinical imaging ...

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Tavşanlarda Subretinal Cerrahi Adım Protokolle Bir Adım

Microsurgical Dissection and Tissue Clearing for High Resolution Intact Whole Retina and Vitreous Imaging

Kemirgenlerde Cerrahi Olarak İndüklenen Glokom için Limbal Vasküler Pleksusun Tam Daire Koterizasyonu

Dejeneratif Retina Hastalıkları için İnsan Dışı Primat Modelinde Retina Pigment Epitel Transplantasyonu

Ex Vivo Yaşa Bağlı Makula Dejenerasyonu Araştırmaları için İnsan Donör Gözlerinin OCT Tabanlı Multimodal Görüntülemesi