A membrane-permeable, UV-light excitable form of Fura-2
Fura-2 AM is a ratiometric fluorescent Ca2+ indicator.
The trypsin (100 nM) mediated Ca2+ transients are imagined in tsA-201 cells loaded with either 750 nM (n=111), 500 nM (n=119) or 250 nM (n=130) Fura-2 AM and compare the average Ca2+ increase above the baseline to the value obtained in the initial experiments using 1 μM of Fura-2 AM. The Ca2+ increases above the baseline for the 750, 500 and 250 nM are 633.0±33.9 nM (n=111, P>0.05 compare to 1 μM Fura-2 AM), 552.6±22.7 nM (n=119, P>0.05 compare to 1 μM Fura-2 AM) and 616.0±24.9 nM (n=130, P>0.05 compare to 1 μM Fura-2 AM), respectively[1].
参考文献:
[1]. Tinning PW, et al. A 340/380 nm light-emitting diode illuminator for Fura-2 AM ratiometric Ca2+ imaging of live cells with better than 5 nM precision. J Microsc. 2017 Aug 24.
[2]. Odmara L Barreto-Chang, et al. Calcium Imaging of Cortical Neurons using Fura-2 AM. J Vis Exp. 2009 Jan 19;(23). pii: 1067.
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Cell experiment: |
Cells are loaded with Fura-2 AM, which diffuses across the cell membrane and is de-esterified by cellular esterases to yield Fura-2 free acid. Aliquot 2-mL of culture media into a 15 mL conical tube, warm to 37 deg. and add 2 μL of Fura-2 AM stock to generate a 1 μM Fura-2 AM solution. Vortex the solution vigorously for 1 minute. Transfer the loading solution to a 35 mm tissue culture dish and transfer the coverslip with the cells into the dish. Incubate the neurons at 37 degrees for 30 minutes in a dark incubator. Prepare a 35 mm dish containing 2-mL of tissue culture media without Fura-2 AM. Remove the coverslip from the loading solution and place in the new dish. Mount the coverslip on the imaging chamber[2]. |
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参考文献: [1]. Tinning PW, et al. A 340/380 nm light-emitting diode illuminator for Fura-2 AM ratiometric Ca2+ imaging of live cells with better than 5 nM precision. J Microsc. 2017 Aug 24. |
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