Abstract
Nuclear Ca2+ regulates key cellular processes, including gene expression, apoptosis, assembly of the nuclear envelope, and nucleocytoplasmic transport. Quantifi cation of subcellularly resolved Ca2+ signals is, therefore, essential for understanding physiological and pathological processes in various cell types. However, the properties of commonly used Ca2+ -fl uorescent indicators in intracellular compartments may differ, thus affecting the translation of Ca2+ -dependent fl uorescence changes into quantitative changes of Ca2+ concentration. Here, we describe technical approaches for reliable subcellular quantifi cation of [Ca2+] in the cytoplasm vs. the nucleus and the nuclear envelope by in situ calibration of fl uorescein-derived fl uorescent indicators Fluo-4 and Fluo-5N.
Original language | English (US) |
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Pages (from-to) | 135-147 |
Number of pages | 13 |
Journal | Methods in Molecular Biology |
Volume | 1234 |
DOIs | |
State | Published - 2015 |
Keywords
- Ca concentration
- Cytoplasm
- Fluo-4
- Fluo-5N
- In situ calibration
- Nuclear envelope
- Nucleus
ASJC Scopus subject areas
- Molecular Biology
- Genetics