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A highly sensitive fluorescent indicator dye for calcium imaging of neural activity in vitro and in vivo.

Tada M, Takeuchi A, Hashizume M, Kitamura K, Kano M - Eur. J. Neurosci. (2014)

Bottom Line: Therefore, it is difficult to detect signals caused by single action potentials (APs) particularly from neurons in vivo.Here we showed that a recently developed calcium indicator dye, Cal-520, is sufficiently sensitive to reliably detect single APs both in vitro and in vivo.These characteristics of Cal-520 are a great advantage over those of Oregon Green BAPTA-1, the most commonly used calcium indicator dye, for monitoring the activity of individual neurons both in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurophysiology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.

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Time course of the change in fluorescence signals by Cal-520. (A) Fluorescence images of layer 2/3 pyramidal neurons in the barrel cortex recorded at 0.5 h (left) and 4.5 h (right) after the injection of Cal-520. Scale bars, 20 μm. Bar graphs showing the mean baseline fluorescence (B), amplitude (C) and frequency (D) of calcium transients by Cal-520 recorded within 1.0 h or more than 4 h after the injection. ***P=0.00002.
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fig06: Time course of the change in fluorescence signals by Cal-520. (A) Fluorescence images of layer 2/3 pyramidal neurons in the barrel cortex recorded at 0.5 h (left) and 4.5 h (right) after the injection of Cal-520. Scale bars, 20 μm. Bar graphs showing the mean baseline fluorescence (B), amplitude (C) and frequency (D) of calcium transients by Cal-520 recorded within 1.0 h or more than 4 h after the injection. ***P=0.00002.

Mentions: The time course of the change in fluorescence signals by Cal-520 was examined. The mean baseline fluorescence, amplitude, and frequency of spontaneous calcium transients in neurons were compared at different time points during imaging (Fig.6). During 1–4 h after the injection, Cal-520 showed no significant change in the mean amplitude of calcium transients (0.325 ± 0.003 and 0.291 ± 0.009 ΔF/F, P= 0.49, t-test) (Fig.6C), despite an increase of baseline fluorescence intensity (52.8 ± 0.3 and 73.9 ± 0.8 F in 59 cells and 22 cells, respectively, P=0.00002, t-test) (Fig.6B). The frequency of spontaneous calcium transients was also unchanged (0.0457 ± 0.0006 and 0.0391 ± 0.011 Hz, P=0.36, t-test) (Fig.6D).


A highly sensitive fluorescent indicator dye for calcium imaging of neural activity in vitro and in vivo.

Tada M, Takeuchi A, Hashizume M, Kitamura K, Kano M - Eur. J. Neurosci. (2014)

Time course of the change in fluorescence signals by Cal-520. (A) Fluorescence images of layer 2/3 pyramidal neurons in the barrel cortex recorded at 0.5 h (left) and 4.5 h (right) after the injection of Cal-520. Scale bars, 20 μm. Bar graphs showing the mean baseline fluorescence (B), amplitude (C) and frequency (D) of calcium transients by Cal-520 recorded within 1.0 h or more than 4 h after the injection. ***P=0.00002.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4232931&req=5

fig06: Time course of the change in fluorescence signals by Cal-520. (A) Fluorescence images of layer 2/3 pyramidal neurons in the barrel cortex recorded at 0.5 h (left) and 4.5 h (right) after the injection of Cal-520. Scale bars, 20 μm. Bar graphs showing the mean baseline fluorescence (B), amplitude (C) and frequency (D) of calcium transients by Cal-520 recorded within 1.0 h or more than 4 h after the injection. ***P=0.00002.
Mentions: The time course of the change in fluorescence signals by Cal-520 was examined. The mean baseline fluorescence, amplitude, and frequency of spontaneous calcium transients in neurons were compared at different time points during imaging (Fig.6). During 1–4 h after the injection, Cal-520 showed no significant change in the mean amplitude of calcium transients (0.325 ± 0.003 and 0.291 ± 0.009 ΔF/F, P= 0.49, t-test) (Fig.6C), despite an increase of baseline fluorescence intensity (52.8 ± 0.3 and 73.9 ± 0.8 F in 59 cells and 22 cells, respectively, P=0.00002, t-test) (Fig.6B). The frequency of spontaneous calcium transients was also unchanged (0.0457 ± 0.0006 and 0.0391 ± 0.011 Hz, P=0.36, t-test) (Fig.6D).

Bottom Line: Therefore, it is difficult to detect signals caused by single action potentials (APs) particularly from neurons in vivo.Here we showed that a recently developed calcium indicator dye, Cal-520, is sufficiently sensitive to reliably detect single APs both in vitro and in vivo.These characteristics of Cal-520 are a great advantage over those of Oregon Green BAPTA-1, the most commonly used calcium indicator dye, for monitoring the activity of individual neurons both in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurophysiology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.

Show MeSH
Related in: MedlinePlus