Biophysical Journal
Volume 117, Issue 10, 19 November 2019, Pages 1783-1794
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Article
Time-Resolved Integrative Optical Imaging of Diffusion during Spreading Depression

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Abstract

An improved version of the integrative optical imaging method has been developed that substantially increases the time resolution of diffusion measurements. We present a theory for time-resolved integrative optical imaging that incorporates a time-dependent effective diffusion coefficient in homogeneous anisotropic media and a time-dependent nonspecific linear clearance. The method was applied to measure the very fast changes in extracellular diffusion that occur during spreading depression in rat hippocampal slices. We were able to achieve a time resolution of approximately 1 s, an improvement of at least 10 times compared to the standard methods for extracellular diffusion measurement. We have found that diffusion of a small fluorescent extracellular marker (MW 3000) completely stopped during the maximum direct current shift associated with the spreading depression wave, then gradually resumed over several minutes afterward. The effect of spreading depression on extracellular space is much larger than previously estimated by other methods with lower time resolution.

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