Publication Date:
2022-10-27
Description:
© The Author(s), 2019. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Frontiers in Cellular Neuroscience 12 (2019): 514. doi: 10.3389/fncel.2018.00514.
Description:
High speed imaging of ion concentration changes in neurons is an important and growing tool for neuroscientists. We previously developed a system for simultaneously measuring sodium and calcium changes in small compartments in neurons (Miyazaki and Ross, 2015). We used this technique to analyze the dynamics of these ions in individual pyramidal neuron dendritic spines (Miyazaki and Ross, 2017). This system is based on high speed multiplexing of light emitting diodes (LEDs) and classic organic indicators. To improve this system we made additional changes, primarily incorporating lasers in addition to the LEDs, more sophisticated imaging protocols, and the use of newer sodium and calcium indicators. This new system generates signals with higher signal to noise ratio (S/N), less background fluorescence, and less photodynamic damage. In addition, by using longer wavelength indicators instead of indicators sensitive in the UV range, it allows for the incorporation of focal uncaging along with simultaneous imaging, which should extend the range of experiments.
Description:
This work was supported in part by National Institutes of Health Grants R21NS085729 (WR), R01NS099122 (WR), and R01NS103168 (JL).
Keywords:
Sodium
;
Calcium
;
Imaging
;
Dendrite
;
Spine
;
Pyramidal neuron
;
CCD camera
Repository Name:
Woods Hole Open Access Server
Type:
Article
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