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Massachusetts General Hospital–Harvard Medical School–Shriners Burns Hospital

3 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Infection of Zebrafish Embryos with Intracellular Bacterial Pathogens

Erica L. Benard¹, Astrid M. van der Sar², Felix Ellett³, Graham J. Lieschke³, Herman P. Spaink¹, Annemarie H. Meijer¹

¹Department of Molecular Cell Biology, Institute of Biology, Leiden University, ²Department of Medical Microbiology and Infection Control, VU University Medical Center, ³Australian Regenerative Medicine Institute, Monash University

Transparent zebrafish embryos have proved useful model hosts to visualize and functionally study interactions between innate immune cells and intracellular bacterial pathogens, such as Salmonella typhimurium and Mycobacterium marinum. Micro-injection of bacteria and multi-color fluorescence imaging are essential techniques involved in the application of zebrafish embryo infection models.

Bioengineering

Microfluidic Platform for Measuring Neutrophil Chemotaxis from Unprocessed Whole Blood

Caroline N. Jones^*1,2,3, Anh N. Hoang^*1,2,3, Laurie Dimisko¹, Bashar Hamza¹, Joseph Martel^1,4, Daniel Irimia^1,2,3

¹The BioMEMS Resource Center, Department of Surgery, Massachusetts General Hospital, ²Harvard Medical School, ³Shriners Burns Hospital, ⁴Harvard University School of Engineering and Applied Sciences

This protocol details an assay designed to measure human neutrophil chemotaxis from one droplet of whole blood with robust reproducibility. This approach circumvents the need for neutrophil separation and requires only a few minutes of assay preparation time. The microfluidic chip enables the repeated measure of neutrophil chemotaxis over time in infants or small mammals, where sample volume is limited.

Developmental Biology

Microstructured Devices for Optimized Microinjection and Imaging of Zebrafish Larvae

Felix Ellett¹, Daniel Irimia¹

¹BioMEMS Resource Center, Department of Surgery, Massachusetts General Hospital–Harvard Medical School–Shriners Burns Hospital

Microinjection of zebrafish embryos and larvae is a crucial but challenging technique used in many zebrafish models. Here, we present a range of microscale tools to aid in the stabilization and orientation of zebrafish for both microinjection and imaging.