In order to identify potential of neuronal differentiation, we evaluated calcium influx which is an indicator for neurotransmitter transmission. The intracellular calcium assessment was described in previous study [23 ]. The specimens were incubated with 3 μM Fluo-3 AM (Invitrogen) and 0.08% pluronic acid (Invitrogen) in DMEM/F-12, 100 U/mL penicillin, and 100 μM/mL streptomycin at 37 °C for 60 min. Subsequently, the specimens were washed with DMEM/F-12, 100 U/mL penicillin, 100 μM/mL streptomycin, and PBS. The specimens were maintained in Tyrode’s solution (5 mM KCl, 129 mM NaCl, 2 mM CaCl2, 1 mM MgCl2, 30 mM glucose, and 25 mM HEPES, pH 7.4) (all from Sigma-Aldrich). The neurotransmitter releasing ability of differentiated cells was simulated with 50 mM KCI. The intensity of calcium was recorded time-lapse at excitation 506 nm for 3 min by the live-cell fluorescence microscope, IX83XDC (Olympus) and interpreted using the ImageJ program (NIH).
Ix83xdc
Manufactured by Olympus
The IX83XDC is an inverted research microscope designed for advanced imaging applications. It features a sturdy and stable frame, high-precision optics, and a modular design to accommodate a variety of accessories. The IX83XDC is suitable for a range of microscopy techniques, including fluorescence, phase contrast, and brightfield imaging.
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3 protocols using ix83xdc
1
Evaluating Neuronal Differentiation via Calcium Influx
2
Measuring Neuronal Calcium Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
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In order to identify potential of neuronal differentiation, we evaluated calcium in ux which is an indicator for neurotransmitter transmission. The intracellular calcium assessment was described in previous study (23) . The specimens were incubated with 3 µM Fluo-3 AM (Invitrogen) and 0.08% pluronic acid (Invitrogen) in DMEM/F-12, 100 U/mL Penicillin, 100 μM/mL Streptomycin at 37 o C for 60 minutes. Subsequently, the specimens were washed with DMEM/F-12, 100 U/mL Penicillin, 100 μM/mL Streptomycin, and PBS. The specimens were maintained in Tyrode's solution (5 mM KCl, 129 mM NaCl, 2 mM CaCl 2 , 1 mM MgCl 2 , 30 mM glucose, and 25 mM HEPES, pH 7.4) (All from Sigma-Aldrich). The neurotransmitter releasing ability of differentiated cells was simulated with 50 mM KCI. The intensity of calcium was recorded time-lapse at excitation 506 nm for 3 minutes by the Live-cell uorescence microscope, IX83XDC (Olympus), and interpreted using the ImageJ program (NIH).
3
Evaluating Neuronal Calcium Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In order to identify potential of neuronal differentiation, we evaluated calcium in ux which is an indicator for neurotransmitter transmission. The intracellular calcium assessment was described in previous study (23) . The specimens were incubated with 3 µM Fluo-3 AM (Invitrogen) and 0.08% pluronic acid (Invitrogen) in DMEM/F-12, 100 U/mL Penicillin, 100 μM/mL Streptomycin at 37 o C for 60 minutes. Subsequently, the specimens were washed with DMEM/F-12, 100 U/mL Penicillin, 100 μM/mL Streptomycin, and PBS. The specimens were maintained in Tyrode's solution (5 mM KCl, 129 mM NaCl, 2 mM CaCl 2 , 1 mM MgCl 2 , 30 mM glucose, and 25 mM HEPES, pH 7.4) (All from Sigma-Aldrich). The action potential ability of differentiated cells was simulated with 50 mM KCI. The intensity of calcium was recorded time-lapse at excitation 506 nm for 3 minutes by the Live-cell uorescence microscope, IX83XDC (Olympus), and interpreted using the ImageJ program (NIH).
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