It was a pleasure to present this poster at the FENS Forum. The poster details Metrion and Scientifica’s¹ collaboration on the optimisation and utilisation of the PatchScope Pro². We first targeted three different TRP channels (TRPV1, TRPA1 and TRPM8) in rat dorsal root ganglia (DRGs). Subsequently we focused on TRPM8 which is a multimodal sensor (temperature, pressure, voltage, osmolality) present in 10-20% of DRGs neurons.

Using calcium-imaging, response of DRG neurons to TRPV1, TRPA1 and TRPM8 ligands were tested, where 64% of neurons responded to 10 µM capsaicin, 29% to 30 µM cinnamaldehyde and 13% to 500 µM menthol. Subsequently a compound-application protocol was optimized using repeat applications of 200 µM menthol, suitable for combined calcium-imaging/electrophysiology. TRPM8 expressing neurons were characterized using electrophysiology following initial selection by calcium-imaging (menthol positive), where effects of 200 µM menthol were demonstrated in both voltage-clamp and current-clamp recordings (n=5). This study demonstrates the advantages of the PatchScope system for selecting specific subsets of neurons using calcium-imaging prior to electrophysiology recordings.

1. 1. https://www.scientifica.uk.com.

1. 1. Scientifica’s PatchScope Pro is an integrated electrophysiology rig incorporating an inverted phase-contrast fluorescence microscope, motorised XY stage and PatchStar micromanipulators, suitable for patch-clamp recording. In contrast to other patch-clamp systems, this microscope has a small footprint to maximise use of lab space. The low-noise electrical components are designed specifically for patch-clamp recordings. Fluorescence capability allows identification of cells of interest (e.g. using fluorescent markers or calcium-imaging) and motorisation of the stage means that XY co-ordinates of cells can be stored for subsequent electrophysiology and imaging.

It was a pleasure to present this poster at the FENS Forum. The poster details Metrion and Scientifica’s¹ collaboration on the optimisation and utilisation of the PatchScope Pro². We first targeted three different TRP channels (TRPV1, TRPA1 and TRPM8) in rat dorsal root ganglia (DRGs). Subsequently we focused on TRPM8 which is a multimodal sensor (temperature, pressure, voltage, osmolality) present in 10-20% of DRGs neurons.

Using calcium-imaging, response of DRG neurons to TRPV1, TRPA1 and TRPM8 ligands were tested, where 64% of neurons responded to 10 µM capsaicin, 29% to 30 µM cinnamaldehyde and 13% to 500 µM menthol. Subsequently a compound-application protocol was optimized using repeat applications of 200 µM menthol, suitable for combined calcium-imaging/electrophysiology. TRPM8 expressing neurons were characterized using electrophysiology following initial selection by calcium-imaging (menthol positive), where effects of 200 µM menthol were demonstrated in both voltage-clamp and current-clamp recordings (n=5). This study demonstrates the advantages of the PatchScope system for selecting specific subsets of neurons using calcium-imaging prior to electrophysiology recordings.

1. 1. https://www.scientifica.uk.com.

1. 1. Scientifica’s PatchScope Pro is an integrated electrophysiology rig incorporating an inverted phase-contrast fluorescence microscope, motorised XY stage and PatchStar micromanipulators, suitable for patch-clamp recording. In contrast to other patch-clamp systems, this microscope has a small footprint to maximise use of lab space. The low-noise electrical components are designed specifically for patch-clamp recordings. Fluorescence capability allows identification of cells of interest (e.g. using fluorescent markers or calcium-imaging) and motorisation of the stage means that XY co-ordinates of cells can be stored for subsequent electrophysiology and imaging.