What are they used for?
These restraints may be used for any small tissue that would be difficult to adhere to the surface of a 96-well plate for analysis. While this restraint was designed with the intent of analyzing metabolism, it may be used for any application where immobilizing tissues without physically adhering them would be useful.
These were initially developed to assist the D. melanogaster community’s investigation of metabolism in neurological disease models using the XFe96 Flux Analyzer to analyze the fly brain. Measuring metabolism of whole brains with this instrument is challenging due to the geometry of the brain and the analyzers requirement for samples to remain at the bottom of a 96-well plate. Thus this requires a specially designed and manufactured micro-tissue restraint that circumvents this problem by preventing movement of the brain while still allowing metabolic measurements from the analyzers two solid-state sensor probes.
With minor optimization this protocol can be adapted for use with any whole tissue and/or model system, provided that the sample size does not exceed the chamber generated by the restraint. Countless experimental conditions, such as energy source preference, interrogation of specific metabolic pathways, and rearing environment, can be interrogated.
About the Inventors
Marla Tipping, Ph.D.
Marla Tipping graduated from Saint Anselm College with a B.A. in Biology. She earned her doctoral degree from the University of Massachusetts Boston in Cellular, Molecular and Organismal Biology, and completed a Ruth L. Kirschstein-National Research Service Award postdoctoral fellowship at Harvard Medical School in the lab of Norbert Perrimon.
During her postdoc she became interested in studying shifts in metabolism during disease and development, and has continued this work at Providence College where she is currently an associate professor investigating metabolic reprogramming in neurodegenerative disease. Dr. Tipping has over 15 years of experience genetically manipulating fruit flies for cellular, molecular and metabolic analysis.
James Waters, Ph.D.
James S. Waters earned an A.B. in Mathematics at The University of Chicago, a Ph.D. in Biology at Arizona State University, and was a James S. McDonnell Foundation postdoctoral fellow at Princeton University prior to joining the faculty in the Department of Biology at Providence College where he is currently an associate professor.
Dr. Waters has co-authored 21 peer-reviewed research papers and one book chapter and has experience designing and machining respirometry chambers for precise measurements of insect metabolic rates.
Neville, K. E., Bosse, T. L., Klekos, M., Mills, J. F., Tipping, M. Metabolic Analysis of Drosophila melanogaster Larval and Adult Brains. J. Vis. Exp. (138), e58007, doi:10.3791/58007 (2018).
Journal of Neuroscience Methods:
Neville, K. E., Bosse, T. L., Klekos, M., Mills, J. F., Weicksel, S. E., Waters, J. S., & Tipping, M. (2018). A novel ex vivo method for measuring whole brain metabolism in model systems. Journal of neuroscience methods, 296, 32–43. https://doi.org/10.1016/j.jneumeth.2017.12.020