Enhanced Loading of Fura-2/AM Calcium Indicator Dye in Adult Rodent Brain Slices Via a Microfluidic Oxygenator
journal contributionposted on 2016-03-28, 00:00 authored by Gerardo Mauleon, Joe F. Lo, Bethany L. Peterson, Christopher P. Fall, David T. Eddington
A microfluidic oxygenator is used to deliver constant oxygen to rodent brain slices, enabling the loading of the cell-permeant calcium indicator Fura-2/AM into cells of adult brain slices. When compared to traditional methods, our microfluidic oxygenator improves loading efficiency, measured by the number of loaded cells per unit area, for all tested age groups. Loading in slices from 1-year-old mice was achieved, which has not been possible with current bulk loading methods. This technique significantly expands the age range for which calcium studies are possible without cellular injection. This technique will facilitate opportunities for the study of calcium signaling of aging and long term stress related diseases. Moreover, it should be applicable to other membrane-permeant physiological indicator varieties.
The authors are grateful for the support of the NIMH Neurotechnology program through R21MH085073 and Chicago Biomedical Consortium with support from the Searle Funds at The Chicago Community Trust.
Publisher StatementNOTICE: This is the author’s version of a work that was accepted for publication in Journal of Neuroscience Methods. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Journal of Neuroscience Methods, Vol 216, Issue 2, 2013 DOI:10.1016/j.jneumeth.2013.04.007