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J Neurosci Methods. 2013 Jun 15;216(2):110-7. doi: 10.1016/j.jneumeth.2013.04.007. Epub 2013 Apr 19.

Enhanced loading of Fura-2/AM calcium indicator dye in adult rodent brain slices via a microfluidic oxygenator.

Author information

1
Department of Bioengineering, University of Illinois at Chicago, USA.

Abstract

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.

PMID:
23608309
PMCID:
PMC3767143
DOI:
10.1016/j.jneumeth.2013.04.007
[Indexed for MEDLINE]
Free PMC Article

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