Cited by 416Advanced Fluorescence Microscopy TechniquesIon Channels and ReceptorsNeuroscience and Neuropharmacology Research

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Estimating Intracellular Calcium Concentrations and Buffering without Wavelength Ratioing

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Maravall, M., Mainen, Z.F., Sabatini, B.L., Svoboda, K.·2000·Biophysical Journal

DOI 10.1016/s0006-3495(00)76809-3PMC PMC1300854

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Abstract

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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A new generation of Ca2+ indicators with greatly improved fluorescence properties.
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Authors: Maravall, M., Mainen, Z. F., Sabatini, B. L., & Svoboda, K.
Affiliations: Cold Spring Harbor Laboratory (US)
Corresponding author: Svoboda, K.
ORCID: Maravall, M.
Mainen, Z.F.
Sabatini, B.L.
Svoboda, K.
Venue: Biophysical Journal
Publisher: Elsevier BV
Published: 2000-05
Volume: 78
Issue: 5
Pages: 2655-2667
ISSN: 0006-3495 (ISSN-L)
DOI: 10.1016/s0006-3495(00)76809-3
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: www.elsevier.com/tdm/userlicense/1.0/ (tdm)
www.elsevier.com/open-access/userlicense/1.0/ (vor, 4824 d)
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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Estimating Intracellular Calcium Concentrations and Buffering without Wavelength Ratioing

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