National Eye Institute

7 ARTICLES PUBLISHED IN JoVE

Neuroscience

An Optic Nerve Crush Injury Murine Model to Study Retinal Ganglion Cell Survival

Zhongshu Tang¹, Shuihua Zhang^1,2, Chunsik Lee¹, Anil Kumar¹, Pachiappan Arjunan¹, Yang Li¹, Fan Zhang¹, Xuri Li¹

¹National Eye Institute, NIH, ²Ophthalmology Department, The Second Hospital of Harbin Medical University

This protocol shows how to retrogradely label retinal ganglion cells, and how to subsequently make an optic nerve crush injury in order to analyze retinal ganglion cell survival and apoptosis. It is an experimental disease model for different types of optic neuropathy, including glaucoma.

Medicine

A Mouse Model of the Cornea Pocket Assay for Angiogenesis Study

Zhongshu Tang¹, Fan Zhang¹, Yang Li¹, Pachiappan Arjunan¹, Anil Kumar¹, Chunsik Lee¹, Xuri Li¹

¹National Eye Institute

The cornea is unique in that it lacks vascular tissues. However, robust blood vessel growth and survival can be induced in the cornea by potent angiogenic factors. Therefore, the cornea can provide with us a valuable tool for angiogenic studies. This protocol demonstrates how to perform the mouse model of cornea pocket assay and how to assess the angiogenesis induced by angiogenic factors using this model.

Bioengineering

Imaging Denatured Collagen Strands In vivo and Ex vivo via Photo-triggered Hybridization of Caged Collagen Mimetic Peptides

Yang Li¹, Catherine A. Foss², Martin G. Pomper^2,3, S. Michael Yu^1,3

¹Department of Bioengineering, University of Utah, ²Department of Radiology and Radiological Science, Johns Hopkins University School of Medicine, ³Institute for NanoBiotechnology, Johns Hopkins University

This procedure demonstrates in vivo near IR fluorescence imaging of collagen remodeling activities in mice as well as ex vivo staining of collagens in tissue sections using caged collagen mimetic peptides that can be photo-triggered to hybridize with denatured collagen strands.

Bioengineering

Engineering Platform and Experimental Protocol for Design and Evaluation of a Neurally-controlled Powered Transfemoral Prosthesis

Fan Zhang¹, Ming Liu¹, Stephen Harper^2,3, Michael Lee³, He Huang¹

¹Joint Department of Biomedical Engineering, North Carolina State University & University of North Carolina at Chapel Hill, ²Department of Physical Medicine and Rehabilitation, University of North Carolina School of Medicine, ³Atlantic Prosthetics & Orthotics, LLC

Neural-machine interfaces (NMI) have been developed to identify the user's locomotion mode. These NMIs are potentially useful for neural control of powered artificial legs, but have not been fully demonstrated. This paper presented (1) our designed engineering platform for easy implementation and development of neural control for powered lower limb prostheses and (2) an experimental setup and protocol in a laboratory environment to evaluate neurally-controlled artificial legs on patients with lower limb amputations safely and efficiently.

Immunology and Infection

User-friendly, High-throughput, and Fully Automated Data Acquisition Software for Single-particle Cryo-electron Microscopy

Anil Kumar¹, Surekha P.¹, Sahil Gulati², Somnath Dutta¹

¹Molecular Biophysics Unit, Indian Institute of Science, ²Gatan Inc.

Single-particle cryo-electron microscopy demands a suitable software package and user-friendly pipeline for high-throughput automatic data acquisition. Here, we present the application of a fully automated image acquisition software package, Latitude-S, and a practical pipeline for data collection of vitrified biomolecules under low-dose conditions.

Medicine

Isolation of Monocyte-Macrophage Lineage Cells from Rat Bones by Secondary Adherence Method

Xiaoli Jin^*1, Yang Li^*2, Xuanwei Chen¹, Jin Chen¹, Jian Xu¹

¹School of Medical Technology and Information Engineering, Zhejiang Chinese Medical University, ²School of Basic Medical Sciences, Fudan University

Here we present a protocol for the isolation of BMMs from SD rats, called the secondary adherence method.

Immunology and Infection

Efficient Transfection of In vitro Transcribed mRNA in Cultured Cells Using Peptide-Poloxamine Nanoparticles

Qin Xiao^*1, Yuheng Liu^*1, Dandan Zhang¹, Chao Li¹, Qihua Yang¹, Dongshui Lu¹, Weijun Zhang¹, Joseph Rosenecker², Quanming Zou¹, Yang Li³, Shan Guan¹

¹National Engineering Research Center of Immunological Products, Department of Microbiology and Biochemical Pharmacy, Third Military Medical University, ²Department of Pediatrics, Ludwig-Maximilians University of Munich, ³Department of Pharmacy, Southwest Hospital, Third Military Medical University

A self-assembled peptide-poloxamine nanoparticle (PP-sNp) is developed using a microfluidic mixing device to encapsulate and deliver in vitro transcribed messenger RNA. The described mRNA/PP-sNp could efficiently transfect cultured cells in vitro.