US Food and Drug Administration

3 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

High-throughput Quantitative Real-time RT-PCR Assay for Determining Expression Profiles of Types I and III Interferon Subtypes

Lynnsey A. Renn¹, Terence C. Theisen¹, Maria B. Navarro¹, Viraj P. Mane¹, Lynnsie M. Schramm¹, Kevin D. Kirschman¹, Giulia Fabozzi¹, Philippa Hillyer¹, Montserrat Puig², Daniela Verthelyi², Ronald L. Rabin¹

¹Center for Biologics Evaluation and Research, US Food and Drug Administration, ²Center for Drug Evaluation and Research, US Food and Drug Administration

This protocol describes a high-throughput qRT-PCR assay for the analysis of type I and III IFN expression signatures. The assay discriminates single base pair differences between the highly similar transcripts of these genes. Through batch assembly and robotic pipetting, the assays are consistent and reproducible.

JoVE Journal

Live Cell Imaging of Chromosome Segregation During Mitosis

Prajakta Varadkar¹, Kazuyo Takeda², Brenton McCright¹

¹Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, US Food and Drug Administration, ²Microscopy and Imaging Core facility, Division of Viral Products, Center for Biologics Evaluation and Research, US Food and Drug Administration

This protocol describes an easy and convenient method to label and visualize live chromosomes in mitotic cells using Histone2B-GFP BacMam 2.0 label and a spinning disc confocal microscopy system.

Bioengineering

Earthworm, Lumbricus Terrestris: A Novel Microinjection Vasculature In vivo Invertebrate Model

Asis Lopez^1,5, Yaswitha Mikkilineni², Shayna Berman³, Damir Khismatullin^1,4, Gregory T. Clement⁵, Matthew R. Myers⁵

¹Bioinnovation PhD Program, Tulane University, ²Biology Department, Virginia Commonwealth University, ³Mechanical Engineering Department, Montgomery College, ⁴Biomedical Engineering Department, Tulane University, ⁵US Food and Drug Administration

Earthworms are a novel invertebrate in vivo bench-top model for vasculature studies. We present techniques and equipment that allow efficient surgery and microinjection into the earthworm vasculature. Surgical protocols, microinjection techniques and the procedure for producing custom-made micropipettes are described.