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Calcium Imaging
Published: Apr 5, 2012 DOI: 10.21769/BioProtoc.200 Views: 35825
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Abstract
This is a protocol of Ca2+ imaging experiment using Ca2+ indicator Fura-2. Ca2+ imaging is an efficient and quantitative method for measuring cytosolic and internal store Ca2+ levels, as well as their dynamic changes.
Materials and Reagents
- HEK293 cells
- Phosphate buffered saline (PBS)
- Fura-2-AM (Life Technologies, Invitrogen™, catalog number: F1221 )
- Poly-L-lysine (Sigma-Aldrich, catalog number: P8920 )
- Fura-2 calcium imaging calibration kit (Life Technologies, Invitrogen™, catalog number: F6774 )
- NaCl
- MgCl2
- KCl
- Glucose
- HEPES
- CaCl2
- DMSO
- GFP reporter
- Lonomycin
- Ca2+ recording buffer (see Recipes)
Equipment
- Zeiss inverted microscopy with perfusion system and “IPlab” software.
- Centrifuges
Procedure
Category
Cell Biology > Cell-based analysis > Ion analysis
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