Imaging:Article Title: Small molecule inhibitors of calcium channel activity and uses thereof
Article Snippet: Dorsal Root Ganglion Neuronal Cultures Dorsal root ganglia (DRG) from all levels were dissected from 100 g female Sprague-Dawley rats using known procedures as described previously (Bellampalli, S. S., Ji, Y., Moutal, A., Cai, S., Wijeratne, E. M. K., Gandini, M. A., Yu, J., Chefdeville, A., Dorame, A., Chew, L. A., Madura, C. L., Luo, S., Molnar, G., Khanna, M., Streicher, J. M., Zamponi, G. W., Gunatilaka, A. .. Calcium Imaging Changes in depolarization-induced calcium influx in rat DRG neurons were determined by loading neurons with 3 mM Fura-2AM for 30 minutes at 37° C. (Cat #F1221; Thermo Fisher, stock solution prepared at 1 mM in DMSO, 0.02% pluronic acid, Cat #P-3000MP; Life Technologies, Carlsbad, CA) as previously described in Bellampalli et al. DRG neurons were incubated overnight with 10 μM and 20 μM of test compounds. .. Dorsal root ganglia were excised and placed in sterile DMEM (Cat #11965; Thermo Fisher Scientific, Waltham, MA).
Article Title: Small molecule inhibitors of calcium channel activity and uses thereof
Article Snippet: DRGs were then collected, trimmed at their roots, and enzymatically digested in 3 mL bicarbonate-free, serum-free, sterile DMEM (Cat #11965, Thermo Fisher Scientific, Waltham, MA) solution containing neutral protease (3.125 mg.ml-1, Cat #LS02104; Worthington, Lakewood, NJ) and collagenase type 1 (5 mg/mL, Cat #LS004194, Worthington, Lakewood, NJ) and incubated for 60 minutes at 37° C. under gentle agitation. .. Calcium Imaging in Acutely Dissociated Dorsal Root Ganglion Neurons Dorsal root ganglion neurons were loaded for 30 minutes at 37° C. with 3 μM Fura-2AM (Cat #F1221, Thermo Fisher, stock solution prepared at 1 mM in DMSO, 0.02% pluronic acid, Cat #P-3000MP, Thermo Fisher) to follow changes in intracellular calcium([Ca2+]c) in a standard bath solution containing 139 mM NaCl, 3 mM KCl, 0.8 mM MgCl2, 1.8 mM CaCl2), 10 mM Na HEPES, pH 7.4, 5 mM glucose exactly as previously described [7] Fluorescence imaging was performed with an inverted microscope, NikonEclipseTi-U (Nikon Instruments Inc., Melville, NY), using objective Nikon Fluor 4× and a Photometrics cooled CCD camera CoolSNAPES2 (Roper Scientific, Tucson, AZ) controlled by NIS Elements software (version 4.20, Nikon Instruments). .. Calcium Imaging in Acutely Dissociated Dorsal Root Ganglion Neurons Dorsal root ganglion neurons were loaded for 30 minutes at 37° C. with 3 μM Fura-2AM (Cat #F1221, Thermo Fisher, stock solution prepared at 1 mM in DMSO, 0.02% pluronic acid, Cat #P-3000MP, Thermo Fisher) to follow changes in intracellular calcium([Ca2+]c) in a standard bath solution containing 139 mM NaCl, 3 mM KCl, 0.8 mM MgCl2, 1.8 mM CaCl2), 10 mM Na HEPES, pH 7.4, 5 mM glucose exactly as previously described [7] Fluorescence imaging was performed with an inverted microscope, NikonEclipseTi-U (Nikon Instruments Inc., Melville, NY), using objective Nikon Fluor 4× and a Photometrics cooled CCD camera CoolSNAPES2 (Roper Scientific, Tucson, AZ) controlled by NIS Elements software (version 4.20, Nikon Instruments).
Incubation:Article Title: Small molecule inhibitors of calcium channel activity and uses thereof
Article Snippet: Dorsal Root Ganglion Neuronal Cultures Dorsal root ganglia (DRG) from all levels were dissected from 100 g female Sprague-Dawley rats using known procedures as described previously (Bellampalli, S. S., Ji, Y., Moutal, A., Cai, S., Wijeratne, E. M. K., Gandini, M. A., Yu, J., Chefdeville, A., Dorame, A., Chew, L. A., Madura, C. L., Luo, S., Molnar, G., Khanna, M., Streicher, J. M., Zamponi, G. W., Gunatilaka, A. .. Calcium Imaging Changes in depolarization-induced calcium influx in rat DRG neurons were determined by loading neurons with 3 mM Fura-2AM for 30 minutes at 37° C. (Cat #F1221; Thermo Fisher, stock solution prepared at 1 mM in DMSO, 0.02% pluronic acid, Cat #P-3000MP; Life Technologies, Carlsbad, CA) as previously described in Bellampalli et al. DRG neurons were incubated overnight with 10 μM and 20 μM of test compounds. .. Dorsal root ganglia were excised and placed in sterile DMEM (Cat #11965; Thermo Fisher Scientific, Waltham, MA).
other:
Inverted Microscopy:Article Title: Stretch, Piezo channels, and store operated calcium entry in developing human airway smooth muscle.
Article Snippet: After exposure to the stretching protocols, the collagen coated membranes from Flexcell plates were cut and placed into 8 well chamber slides containing Hanks balanced salt solution [HBSS (Gibco, 14185-052); 2 mM Ca2+, pH 7.4]. .. The cells were loaded with 5 μM Fura-2 AM (Invitrogen, F1225) for 30min at room temperature in HBSS, and imaged using a Nikon Inverted Microscope (TE2000U) and a 12-bit high sensitivity camera color camera with Nikon NIS Elements software, with excitation wavelengths of 340 nm (when calcium is bound) and 380 nm (when calcium is unbound), and emission detected at 510 nm. ..
Article Title: Small molecule inhibitors of calcium channel activity and uses thereof
Article Snippet: DRGs were then collected, trimmed at their roots, and enzymatically digested in 3 mL bicarbonate-free, serum-free, sterile DMEM (Cat #11965, Thermo Fisher Scientific, Waltham, MA) solution containing neutral protease (3.125 mg.ml-1, Cat #LS02104; Worthington, Lakewood, NJ) and collagenase type 1 (5 mg/mL, Cat #LS004194, Worthington, Lakewood, NJ) and incubated for 60 minutes at 37° C. under gentle agitation. .. Calcium Imaging in Acutely Dissociated Dorsal Root Ganglion Neurons Dorsal root ganglion neurons were loaded for 30 minutes at 37° C. with 3 μM Fura-2AM (Cat #F1221, Thermo Fisher, stock solution prepared at 1 mM in DMSO, 0.02% pluronic acid, Cat #P-3000MP, Thermo Fisher) to follow changes in intracellular calcium([Ca2+]c) in a standard bath solution containing 139 mM NaCl, 3 mM KCl, 0.8 mM MgCl2, 1.8 mM CaCl2), 10 mM Na HEPES, pH 7.4, 5 mM glucose exactly as previously described [7] Fluorescence imaging was performed with an inverted microscope, NikonEclipseTi-U (Nikon Instruments Inc., Melville, NY), using objective Nikon Fluor 4× and a Photometrics cooled CCD camera CoolSNAPES2 (Roper Scientific, Tucson, AZ) controlled by NIS Elements software (version 4.20, Nikon Instruments). .. Calcium Imaging in Acutely Dissociated Dorsal Root Ganglion Neurons Dorsal root ganglion neurons were loaded for 30 minutes at 37° C. with 3 μM Fura-2AM (Cat #F1221, Thermo Fisher, stock solution prepared at 1 mM in DMSO, 0.02% pluronic acid, Cat #P-3000MP, Thermo Fisher) to follow changes in intracellular calcium([Ca2+]c) in a standard bath solution containing 139 mM NaCl, 3 mM KCl, 0.8 mM MgCl2, 1.8 mM CaCl2), 10 mM Na HEPES, pH 7.4, 5 mM glucose exactly as previously described [7] Fluorescence imaging was performed with an inverted microscope, NikonEclipseTi-U (Nikon Instruments Inc., Melville, NY), using objective Nikon Fluor 4× and a Photometrics cooled CCD camera CoolSNAPES2 (Roper Scientific, Tucson, AZ) controlled by NIS Elements software (version 4.20, Nikon Instruments).
Software:Article Title: Stretch, Piezo channels, and store operated calcium entry in developing human airway smooth muscle.
Article Snippet: After exposure to the stretching protocols, the collagen coated membranes from Flexcell plates were cut and placed into 8 well chamber slides containing Hanks balanced salt solution [HBSS (Gibco, 14185-052); 2 mM Ca2+, pH 7.4]. .. The cells were loaded with 5 μM Fura-2 AM (Invitrogen, F1225) for 30min at room temperature in HBSS, and imaged using a Nikon Inverted Microscope (TE2000U) and a 12-bit high sensitivity camera color camera with Nikon NIS Elements software, with excitation wavelengths of 340 nm (when calcium is bound) and 380 nm (when calcium is unbound), and emission detected at 510 nm. ..
Article Title: Small molecule inhibitors of calcium channel activity and uses thereof
Article Snippet: DRGs were then collected, trimmed at their roots, and enzymatically digested in 3 mL bicarbonate-free, serum-free, sterile DMEM (Cat #11965, Thermo Fisher Scientific, Waltham, MA) solution containing neutral protease (3.125 mg.ml-1, Cat #LS02104; Worthington, Lakewood, NJ) and collagenase type 1 (5 mg/mL, Cat #LS004194, Worthington, Lakewood, NJ) and incubated for 60 minutes at 37° C. under gentle agitation. .. Calcium Imaging in Acutely Dissociated Dorsal Root Ganglion Neurons Dorsal root ganglion neurons were loaded for 30 minutes at 37° C. with 3 μM Fura-2AM (Cat #F1221, Thermo Fisher, stock solution prepared at 1 mM in DMSO, 0.02% pluronic acid, Cat #P-3000MP, Thermo Fisher) to follow changes in intracellular calcium([Ca2+]c) in a standard bath solution containing 139 mM NaCl, 3 mM KCl, 0.8 mM MgCl2, 1.8 mM CaCl2), 10 mM Na HEPES, pH 7.4, 5 mM glucose exactly as previously described [7] Fluorescence imaging was performed with an inverted microscope, NikonEclipseTi-U (Nikon Instruments Inc., Melville, NY), using objective Nikon Fluor 4× and a Photometrics cooled CCD camera CoolSNAPES2 (Roper Scientific, Tucson, AZ) controlled by NIS Elements software (version 4.20, Nikon Instruments). .. Calcium Imaging in Acutely Dissociated Dorsal Root Ganglion Neurons Dorsal root ganglion neurons were loaded for 30 minutes at 37° C. with 3 μM Fura-2AM (Cat #F1221, Thermo Fisher, stock solution prepared at 1 mM in DMSO, 0.02% pluronic acid, Cat #P-3000MP, Thermo Fisher) to follow changes in intracellular calcium([Ca2+]c) in a standard bath solution containing 139 mM NaCl, 3 mM KCl, 0.8 mM MgCl2, 1.8 mM CaCl2), 10 mM Na HEPES, pH 7.4, 5 mM glucose exactly as previously described [7] Fluorescence imaging was performed with an inverted microscope, NikonEclipseTi-U (Nikon Instruments Inc., Melville, NY), using objective Nikon Fluor 4× and a Photometrics cooled CCD camera CoolSNAPES2 (Roper Scientific, Tucson, AZ) controlled by NIS Elements software (version 4.20, Nikon Instruments).
Fluorescence:Article Title: Small molecule inhibitors of calcium channel activity and uses thereof
Article Snippet: DRGs were then collected, trimmed at their roots, and enzymatically digested in 3 mL bicarbonate-free, serum-free, sterile DMEM (Cat #11965, Thermo Fisher Scientific, Waltham, MA) solution containing neutral protease (3.125 mg.ml-1, Cat #LS02104; Worthington, Lakewood, NJ) and collagenase type 1 (5 mg/mL, Cat #LS004194, Worthington, Lakewood, NJ) and incubated for 60 minutes at 37° C. under gentle agitation. .. Calcium Imaging in Acutely Dissociated Dorsal Root Ganglion Neurons Dorsal root ganglion neurons were loaded for 30 minutes at 37° C. with 3 μM Fura-2AM (Cat #F1221, Thermo Fisher, stock solution prepared at 1 mM in DMSO, 0.02% pluronic acid, Cat #P-3000MP, Thermo Fisher) to follow changes in intracellular calcium([Ca2+]c) in a standard bath solution containing 139 mM NaCl, 3 mM KCl, 0.8 mM MgCl2, 1.8 mM CaCl2), 10 mM Na HEPES, pH 7.4, 5 mM glucose exactly as previously described [7] Fluorescence imaging was performed with an inverted microscope, NikonEclipseTi-U (Nikon Instruments Inc., Melville, NY), using objective Nikon Fluor 4× and a Photometrics cooled CCD camera CoolSNAPES2 (Roper Scientific, Tucson, AZ) controlled by NIS Elements software (version 4.20, Nikon Instruments). .. Calcium Imaging in Acutely Dissociated Dorsal Root Ganglion Neurons Dorsal root ganglion neurons were loaded for 30 minutes at 37° C. with 3 μM Fura-2AM (Cat #F1221, Thermo Fisher, stock solution prepared at 1 mM in DMSO, 0.02% pluronic acid, Cat #P-3000MP, Thermo Fisher) to follow changes in intracellular calcium([Ca2+]c) in a standard bath solution containing 139 mM NaCl, 3 mM KCl, 0.8 mM MgCl2, 1.8 mM CaCl2), 10 mM Na HEPES, pH 7.4, 5 mM glucose exactly as previously described [7] Fluorescence imaging was performed with an inverted microscope, NikonEclipseTi-U (Nikon Instruments Inc., Melville, NY), using objective Nikon Fluor 4× and a Photometrics cooled CCD camera CoolSNAPES2 (Roper Scientific, Tucson, AZ) controlled by NIS Elements software (version 4.20, Nikon Instruments).
|
Thermo Fisher is a verified supplier 
Thermo Fisher manufactures this product 
