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Related Experiment Videos

Determining calcium concentration in heterogeneous model systems using multiple indicators.

Krzysztof L Hyrc1, Ziemowit Rzeszotnik, Bryan R Kennedy

  • 1Hope Center for Neurological Disorders, Alafi Neuroimaging Laboratory and Department of Neurology, Washington University School of Medicine, St. Louis, MI 63110, USA. hyrck@neuro.wustl.edu

Cell Calcium
|March 23, 2007
PubMed
Summary
This summary is machine-generated.

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Heterogeneous calcium distribution in cells leads to underestimation of intracellular calcium concentrations ([Ca2+]i). Using multiple calcium indicators can accurately estimate calcium levels in different cellular compartments.

Area of Science:

  • Cell Biology
  • Biochemistry
  • Biophysics

Background:

  • Intracellular free calcium concentrations ([Ca2+]i) are crucial for cellular processes.
  • Fluorescence microscopy with calcium indicators is standard for measuring [Ca2+]i.
  • Current methods assume homogeneous calcium distribution, which can lead to errors.

Purpose of the Study:

  • To assess errors in [Ca2+]i determination due to heterogeneous calcium distribution.
  • To evaluate the impact of indicator affinity on accuracy.
  • To develop methods for estimating calcium heterogeneity.

Main Methods:

  • Mathematical and experimental models with distinct 'low' and 'high' calcium compartments.
  • Utilized calcium indicators with varying affinities.

Related Experiment Videos

  • Extended standard methods using multiple indicators with different responses.

    • Main Results:

    • Average fluorescence intensity underestimated actual mean [Ca2+]i in heterogeneous models.
    • Low-affinity indicators provided more accurate [Ca2+]i estimates than high-affinity ones.
    • Two indicators partially characterized two-compartment systems; three or more provided full characterization.

    Conclusions:

    • Uneven calcium distribution significantly underestimates actual [Ca2+]i.
    • Multiple calcium indicators are necessary for accurate assessment of calcium heterogeneity.
    • Novel approaches using multiple indicators can estimate compartmental [Ca2+] and dimensions.