The Neurosciences Institute

2 ARTICLES PUBLISHED IN JoVE

Neuroscience

Neuromodulation and Mitochondrial Transport: Live Imaging in Hippocampal Neurons over Long Durations

David B. Edelman^*1, Geoffrey C. Owens^*1, Sigeng Chen^*1

¹Department of Experimental Neurobiology, The Neurosciences Institute

We describe a protocol that allows imaging of mitochondria in living neurons via fluorescence microscopy over long durations. Imaging over extended periods is accomplished through lentivirus-mediated expression of a mitochondrially targeted fluorescent protein and use of an inexpensive stage-top incubator that was designed and built in our laboratory.

Immunology and Infection

Radial Mobility and Cytotoxic Function of Retroviral Replicating Vector Transduced, Non-adherent Alloresponsive T Lymphocytes

Kate L. Erickson^*1, Michelle J. Hickey^*1, Yuki Kato², Colin C. Malone¹, Geoffrey C. Owens¹, Robert M. Prins^1,2, Linda M. Liau^1,4,5, Noriyuki Kasahara^3,5, Carol A. Kruse^1,4,5

¹Department of Neurosurgery, UCLA David Geffen School of Medicine, ²Department of Molecular and Medical Pharmacology, UCLA David Geffen School of Medicine, ³Department of Medicine, UCLA David Geffen School of Medicine, ⁴Brain Research Institute, UCLA David Geffen School of Medicine, ⁵Jonsson Comprehensive Cancer Center, UCLA David Geffen School of Medicine

We describe a protocol to monitor radial mobility of non-adherent immune cells in vitro using a cell sedimentation manifold/slide apparatus. Cell migration is tracked on monolayers of tumor cells or on extracellular matrix proteins. Examination by light and fluorescence microscopy allows for observation of cell mobility and cytotoxic functionality.