Feb 01, 2024

Public workspaceLive-cell imaging; Calcium imaging with Fura-2

DOI
dx.doi.org/10.17504/protocols.io.261genzqwg47/v1
  • 1UCL Institute of Neurology
Minee L Choi
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DOI: dx.doi.org/10.17504/protocols.io.261genzqwg47/v1
Protocol Citation: Minee L Choi 2024. Live-cell imaging; Calcium imaging with Fura-2. protocols.io https://dx.doi.org/10.17504/protocols.io.261genzqwg47/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 01, 2022
Last Modified: May 31, 2024
Protocol Integer ID: 67954
Keywords: ASAPCRN
Abstract
This protocol describes a method to perform calcium imaging with Fura-2 (live-cell imaging)
Cells are washed with HBSS (Cat No. 14025092, ThermofisherScientific)
Cells are loaded with 5uM of Fura 2-AM (F1221, ThermofisherScientific) in HBSS.
Note
Fura 2-AM: 340/380 ratiometric measurement. Upon binding calcium, Fura-2 exhibits 340 nm and does 380 nm in the rest of the condition.

CCD protocol is set up by selecting a501 nm wheel (both 340 nm and 380 nm wavelength are excited b 501 nm).