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Estimating Intracellular Calcium Concentrations and Buffering without Wavelength Ratioing
Open accessAbstract
We describe a method for determining intracellular free calcium concentration ([Ca(2+)]) from single-wavelength fluorescence signals. In contrast to previous single-wavelength calibration methods, the proposed method does not require independent estimates of resting [Ca(2+)] but relies on the measurement of fluorescence close to indicator saturation during an experiment. Consequently, it is well suited to [Ca(2+)] indicators for which saturation can be achieved under physiological conditions. In addition, the method requires that the indicators have large dynamic ranges. Popular indicators such as Calcium Green-1 or Fluo-3 fulfill these conditions. As a test of the method, we measured [Ca(2+)] in CA1 pyramidal neurons in rat hippocampal slices using Oregon Green BAPTA-1 and 2-photon laser scanning microscopy (BAPTA: 1,2-bis(2-aminophenoxy)ethane-N,N,N', N'-tetraacetic acid). Resting [Ca(2+)] was 32-59 nM in the proximal apical dendrite. Monitoring action potential-evoked [Ca(2+)] transients as a function of indicator loading yielded estimates of endogenous buffering capacity (44-80) and peak [Ca(2+)] changes at zero added buffer (178-312 nM). In young animals (postnatal days 14-17) our results were comparable to previous estimates obtained by ratiometric methods (, Biophys. J. 70:1069-1081), and no significant differences were seen in older animals (P24-28). We expect our method to be widely applicable to measurements of [Ca(2+)] and [Ca(2+)]-dependent processes in small neur…
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“relative to the resting level of intracellular Ca2+ (20–50 nM) (109–111)”
From Synaptic memory and CaMKII (2023) · cites this paper as [110]
In the citing paper
“The unusually high Hill coefficient of ∼6 for CaMKII (108) and the high requirement for Ca2+ (half maximal of ∼1.6 µM at saturating CaM) compared to the resting level of Ca2+ (20–50 nM) (109, 110) ensures maintenance of the synapse specificity and Hebbian nature of LTP (19).”
From Synaptic memory and CaMKII (2023) · cites this paper as [110]
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Metadata
- Authors
- Maravall, M., Mainen, Z. F., Sabatini, B. L., & Svoboda, K.
- Affiliations
- Cold Spring Harbor Laboratory (US)
- Corresponding author
- Svoboda, K.
- Venue
- Biophysical Journal
- Publisher
- Elsevier BV
- Published
- 2000-05
- Volume
- 78
- Issue
- 5
- Pages
- 2655-2667
- ISSN
- 0006-3495 (ISSN-L)
- PMC
- PMC1300854
- OpenAlex
- W1981195606
- Cited by
- 416
- References
- 46
- Citations by year
- 2026: 8 · 2025: 11 · 2024: 18 · 2023: 13 · 2022: 11 · 2021: 10 · 2020: 13 · 2019: 19 · 2018: 25 · 2017: 18 · 2016: 17 · 2015: 23 · 2014: 22 · 2013: 21 · 2012: 31
- FWCI
- 5.44
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Biophysical Journal (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Cold Spring Harbor Laboratory Institutional Repository (Cold Spring Harbor Laboratory) (acceptedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Neuroscience › Sensory Systems › Ion Channels and Receptors
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Keywords
- intracellular calcium concentration, CA1 pyramidal neurons, hippocampal slices, dendritic calcium
- MeSH terms
- Action Potentials, Animals, Biophysics, Buffers, Calcium, Intracellular Fluid, Models, Biological, Pyramidal Cells, Calcium Signaling, Rats, Biophysical Phenomena, In Vitro Techniques
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