Lambda 10 to 2 optical filter change

Dorsal root ganglion neurons were prepared as previously
described^{27 (link)} and
were loaded for 30 minutes at 37°C with 3 μM Fura-2AM (Cat#
F1221, Thermo Fisher, stock solution prepared at 1mM in DMSO, 0.02% pluronic
acid, Cat#P-3000MP, Life technologies) 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^{28 (link)}Fluorescence imaging was performed with an inverted microscope,
NikonEclipseTi-U (Nikon Instruments Inc., Melville,
NY), using objective Nikon Fluor 4X and a Photometrics cooled CCD camera
Cool SNAP ES² (Roper
Scientific, Tucson, AZ) controlled by Nis Elements software (version 4.20,
Nikon Instruments). The excitation light was delivered by a Lambda-LS system
(Sutter Instruments, Novato, CA). The excitation filters (340 ± 5 and
380 ± 7) were controlled by a Lambda 10 to 2 optical filter change
(Sutter Instruments). Fluorescence was recorded through a 505-nm dichroic
mirror at 535 ± 25 nm. To minimize photobleaching and phototoxicity,
the images were taken every ~10 seconds during the time-course of the
experiment using the minimal exposure time that provided acceptable image
quality. The changes in [Ca²⁺]_c were monitored by
following a ratio of F₃₄₀/F₃₈₀, calculated after
subtracting the background from both channels.

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 1mM in DMSO, 0.02% pluronic acid, Cat#P-3000MP, Life Technologies) 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 [8 ]. Fluorescence imaging was performed with an inverted microscope, NikonEclipseTi-U (Nikon Instruments Inc., Melville, NY), using objective Nikon Fluor 4X and a Photometrics cooled CCD camera Cool SNAP ES² (Roper Scientific, Tucson, AZ) controlled by Nis Elements software (version 4.20, Nikon Instruments). The excitation light was delivered by a Lambda-LS system (Sutter Instruments, Novato, CA). The excitation filters (340 ± 5 and 380 ± 7) were controlled by a Lambda 10 to 2 optical filter change (Sutter Instruments). Fluorescence was recorded through a 505-nm dichroic mirror at 535 ± 25 nm. To minimize photobleaching and phototoxicity, the images were taken every ~10 seconds during the time-course of the experiment using the minimal exposure time that provided acceptable image quality. The changes in [Ca²⁺]_c were monitored by following a ratio of F₃₄₀/F₃₈₀, calculated after subtracting the background from both channels.

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 1mM in DMSO, 0.02% pluronic acid, Cat#P-3000MP, Life Technologies) 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 [9 ]. Fluorescence imaging was performed with an inverted microscope, NikonEclipseTi-U (Nikon Instruments Inc., Melville, NY), using objective Nikon Fluor 4X and a Photometrics cooled CCD camera Cool SNAP ES² (Roper Scientific, Tucson, AZ) controlled by Nis Elements software (version 4.20, Nikon Instruments). The excitation light was delivered by a Lambda-LS system (Sutter Instruments, Novato, CA). The excitation filters (340 ± 5 nm and 380 ± 7 nm) were controlled by a Lambda 10 to 2 optical filter change (Sutter Instruments). Fluorescence was recorded through a 505-nm dichroic mirror at 535 ± 25 nm. To minimize photobleaching and phototoxicity, the images were taken every ~10 seconds during the time-course of the experiment using the minimal exposure time that provided acceptable image quality. The changes in [Ca²⁺]_c were monitored by following a ratio of F₃₄₀/F₃₈₀, calculated after subtracting the background from both channels.