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Veterans Affairs Palo Alto Health Care

6 ARTICLES PUBLISHED IN JoVE

Medicine

Dissection of Human Vitreous Body Elements for Proteomic Analysis

¹Department of Ophthalmology and Visual Sciences, Omics Laboratory, University of Iowa

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

Biology

Evisceration of Mouse Vitreous and Retina for Proteomic Analyses

Jessica M. Skeie^1,2, Stephen H. Tsang³, Vinit B. Mahajan^1,2

¹Omics Laboratory, University of Iowa, ²Ophthalmology and Visual Sciences, University of Iowa, ³Harkness Eye Institute, Columbia University College of Physicians and Surgeons

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

Medicine

Mouse Eye Enucleation for Remote High-throughput Phenotyping

Vinit B. Mahajan^1,2, Jessica M. Skeie^1,2, Amir H. Assefnia^2,3, MaryAnn Mahajan^1,2, Stephen H. Tsang^2,4

¹Department of Ophthalmology and Visual Sciences, University of Iowa, ²Omics Laboratory, University of Iowa, ³School of Dentistry, UCLA, ⁴Bernard and Shirlee Brown Glaucoma Laboratory, Department of Ophthalmology, College of Physicians and Surgeons, Columbia University

The dissection technique illustrates enucleation of the mouse eye for tissue fixation to perform phenotyping in high-throughput screens.

Medicine

Subretinal Injection of Gene Therapy Vectors and Stem Cells in the Perinatal Mouse Eye

Katherine J. Wert^1,2, Jessica M. Skeie^3,4, Richard J. Davis¹, Stephen H. Tsang^1,3, Vinit B. Mahajan^3,4

¹Bernard and Shirlee Brown Glaucoma Laboratory, Department of Ophthalmology, Columbia University , ²Institute of Human Nutrition, College of Physicians & Surgeons, Columbia University , ³Omics Laboratory, University of Iowa , ⁴Department of Ophthalmology and Visual Sciences, University of Iowa

This surgical technique illustrates the injection of gene therapy vectors and stem cells into the subretinal space of the mouse eye.

Biochemistry

Dissection of Human Retina and RPE-Choroid for Proteomic Analysis

Thiago Cabral^*1,2,7,8, Marcus A. Toral^*3,4, Gabriel Velez^3,4, James E. DiCarlo^1,2, Anuradha M. Gore³, MaryAnn Mahajan³, Stephen H. Tsang^1,2, Alexander G. Bassuk^5,6, Vinit B. Mahajan^3,9

¹Barbara & Donald Jonas Stem Cell Laboratory, and Bernard & Shirlee Brown Glaucoma Laboratory, Department of Pathology & Cell Biology, Institute of Human Nutrition, College of Physicians and Surgeons, Columbia University, ²Edward S. Harkness Eye Institute, New York-Presbyterian Hospital, ³Omics Laboratory, Byers Eye Institute, Department of Ophthalmology, Stanford University, ⁴Medical Scientist Training Program, University of Iowa, ⁵Department of Pediatrics, University of Iowa, ⁶Department of Neurology, University of Iowa, ⁷Department of Ophthalmology, Federal University of Sao Paulo (UNIFESP), ⁸Department of Ophthalmology, Federal University of EspÍrito Santo (UFES), ⁹Palo Alto Veterans Administration, Palo Alto, CA

The human retina is composed of functionally and molecularly distinct regions, including the fovea, macula, and peripheral retina. Here, we describe a method using punch biopsies and manual removal of tissue layers from a human eye to dissect and collect these distinct retinal regions for downstream proteomic analysis.

JoVE Core

A Reversible Silicon Oil-Induced Ocular Hypertension Model in Mice

Jie Zhang^1,2, Fang Fang^1,3, Liang Li¹, Haoliang Huang¹, Hannah C. Webber¹, Yang Sun^1,4, Vinit B. Mahajan^1,4, Yang Hu¹

¹Department of Ophthalmology, Stanford University School of Medicine, ²Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, ³Department of Ophthalmology, Second Xiangya Hospital of Central South University, ⁴Department of Ophthalmology, Veterans Affairs Palo Alto Health Care

Here, we present a protocol to induce ocular hypertension and glaucomatous neurodegeneration in mouse eyes by intracameral injection of silicone oil and the procedure for silicone oil removal from the anterior chamber to return elevated intraocular pressure to normal.