FLIPR Complete Calibration ( IAFLIPRComplete )

Parameters:

Name	Short Name	Type	Description
Range definitions in time courses	rangedef	string	Defines whether the ranges given below refer to frames or seconds in the recordings
Baseline Range	BaselineRange	string	Defines the range of the baseline (in the above defined time units). Longer baseline results more accurate calibration.
K+ Steps Ranges	KStepsRanges	string	Enter a list of range definitions for the duration of the K+-steps (in the above defined time units), including the MDC treatment.
[K+]ec Steps (mM;mM)	KStepsConcentrations	string	Extracellular (ec) K+ concentrations corresponding to the above defined steps, including MDC, in mM. Use the Wizard tab to calculate concentrations from added volumes.
K+ Step upstroke range	KStepUpstroke	string	Range definitions of a well-visible upstroke for a K+-step (in the above defined time units).
CDC Range	CDCRange	string	The range of Complete Depolarization Cocktail addition (in the above defined time units).
Take the maximum of CDC Range	CDCMax	boolean	If set yes, the maximum FLIPR intensity within the CDC Range is taken as zero calibration value (its error is the local SD of the range). Otherwise mean is calculated.
PN (permeability of plasma membrane to ions other than K+)	PN	real	Summed relative permeability of the plasma membrane to ions other than K+ during * concentration, shown in the numerator of the GHK equation. Use -1 for automatic determination.
fP0-fPX Allowed Range	fP0MinusfPXRange	string	Trace selection criterion: the minimum and maximum allowed value of fP0-fPX
fPX r^2 Min	fpXr2Min	real	Trace selection criterion: the minimum allowed value of r^2 for the FLIPR-regression.
Quality control by propagated error of baseline	QCPMPrest	boolean	Removes traces where the estimated SE of baseline is greater than the threshold below. Requires error propagation.
Maximum SE of baseline (mV)	QCPMPrestMaxSE	real	Traces where the SE of resting ΔψP parameter is larger than this number are removed.
kP Allowed Range	kPRange	string	Trace selection criterion: the minimum and maximum allowed value of kP
Minimum allowed DP/FP	DP/FPMin	real	Criterion for the exact length of data to be used for regression.
kP r^2 Min	kPr2Min	real	Trace selection criterion: the minimum allowed value of r^2 for the FLIPR-regression.

Description:

This method calculates plasma membrane potential (ΔψP) time course by determining all calibration parameters from a single time lapse using the FLIPR Membrane Potential Kit from Molecular Devices (aka PMPI, plasma membrane potential indicator).
Required experimental protocol: any time lapse, followed by establishment of K-equilibrium potential, at least one addition of high K+-medium and finally complete depolarization. Typically use 3-4 K+-steps.
The upstroke of FLIPR signal upon K+-addition has to be well resolved.
The cells have to be able to maintain a constant hyperpolarized ΔψP before starting K+-steps. Do not use this method if ΔψP depolarizes substantially when adding MDC.