This is a summary for all the patch clamp settings in different Piezo1 measurement articles.
Piezo1PatchclampPiezo2
Removal of the mechanoprotective influence of the cytoskeleton reveals PIEZO1 is gated by bilayer tension | Nature Communications
Full article: The activation thresholds and inactivation kinetics of poking-evoked PIEZO1 and PIEZO2 currents are sensitive to subtle variations in mechanical stimulation parameters (tandfonline.com)
| cell model | Neuro2A cells on Poly-L-lysine coated dish |
|---|
| patch type | whole cell |
| intracellular buffer | 125 potassium gluconate, 7 KCl, 1 CaCl2, 1 MgCl2, 10 HEPES, and 4 EGTA (pH 7.3 with KOH), in mM |
| bath solution | 4 KCl, 140 NaCl 1 MgCl2, 2 CaCl2, 4 glucose and 10 HEPES (pH 7.4 with NaOH), in mM |
| holding potential | -60mV |
| sampling frequency | 200kHz with 2.9kHz low pass filter |
| detection threshold | -10pA |
PIEZO1 but not PIEZO2 activates and inactivates faster at higher stimulation angles
PIEZOs inactivate faster when cells are poked with larger glass rods
The mechanical activation threshold of PIEZO2 decreases with increasing stimulation speeds
need to read more about the discussion
Dissecting cell membrane tension dynamics and its effect on Piezo1-mediated cellular mechanosensitivity using force-controlled nanopipettes | Nature Methods
| cell model | HFF cells |
|---|
| patch type | cell attached |
| intracellular buffer | 140 mM NaCl, 5.4 mM KCl, 10 mM HEPES, 10 mM glucose, 1 mM MgCl2, 1.8 mM CaCl2, pH 7.4 with NAOH |
| bath solution | 140 mM NaCl, 5.4 mM KCl, 10 mM HEPES, 10 mM glucose, 1 mM MgCl2, 1.8 mM CaCl2, pH 7.4 with NAOH |
| holding potential | |
| sampling frequency | |
| detection threshold | |
Piezo1 ion channels inherently function as independent mechanotransducers | eLife (elifesciences.org)
| cell model | Neuro2A |
|---|
| patch type | whole cell? |
| intracellular buffer | 130 NaCl, 5 KCl, 10 HEPES, 10 TEACl, 1 CaCl2, 1 MgCl2, and pH 7.3 with NaOH. |
| bath solution | 140 KCl, 10 HEPES, 1 MgCl2, 10 glucose, and pH 7.3 with KOH. |
| holding potential | |
| sampling frequency | |
| detection threshold | |