calibrate_force

lumicks.pylake.calibrate_force

calibrate_force(force_voltage_data, bead_diameter, temperature, *, sample_rate, viscosity=None, active_calibration=False, driving_data=None, driving_frequency_guess=None, axial=False, hydrodynamically_correct=False, rho_sample=None, rho_bead=1060.0, distance_to_surface=None, fast_sensor=False, num_points_per_block=2000, fit_range=(100.0, 23000.0), excluded_ranges=None, fixed_diode=None, fixed_alpha=None, drag=None) → CalibrationResults

Determine force calibration factors.

This function can be used to perform both active and passive calibration.

• For experiments performed near the surface, it is recommended to provide a distance_to_surface.

• For lateral calibration with beads larger than 1 micron, it is recommended to use the hydrodynamically correct theory (hydrodynamically_correct=True), unless so close to the surface (0.75 x diameter) that this model becomes invalid.

• For axial calibration the flag axial=True should be set.

• In the case of a pre-characterized diode, the values for its parameters can be passed to fixed_alpha and fixed_diode.

• In the case of active calibration, it is mandatory to provide a nanostage signal, as well as a guess of the driving frequency.

The power spectrum calibration algorithm implemented here is based on [1] [2] [3] [4] [5] [6]. Please refer to the theory section and tutorial on force calibration for more information on the calibration methods implemented.

Parameters:

• force_voltage_data (numpy.ndarray) – Uncalibrated force data in volts.

• bead_diameter (float) – Bead diameter [um].

• temperature (float) – Liquid temperature [Celsius].

• sample_rate (float) – Sample rate at which the signals were acquired.

• viscosity (float, optional) – Liquid viscosity [Pa*s]. When omitted, the temperature will be used to look up the viscosity of water at that particular temperature.

• active_calibration (bool, optional) – Active calibration, when set to True, driving_data must also be provided.

• driving_data (numpy.ndarray, optional) – Array of driving data.

• driving_frequency_guess (float, optional) – Guess of the driving frequency. Required for active calibration.

• axial (bool, optional) – Is this an axial calibration? Only valid for a passive calibration.

• hydrodynamically_correct (bool, optional) – Enable hydrodynamically correct model.

• rho_sample (float, optional) – Density of the sample [kg/m**3]. Only used when using hydrodynamically correct model.

• rho_bead (float, optional) – Density of the bead [kg/m**3]. Only used when using hydrodynamically correct model.

• distance_to_surface (float, optional) – Distance from bead center to the surface [um] When specifying None, the model will use an approximation which is only suitable for measurements performed deep in bulk.

• fast_sensor (bool, optional) – Fast sensor? Fast sensors do not have the diode effect included in the model.

• fit_range (tuple of float, optional) – Tuple of two floats (f_min, f_max), indicating the frequency range to use for the full model fit. [Hz]

• num_points_per_block (int, optional) – The spectrum is first block averaged by this number of points per block. Default: 2000.

• excluded_ranges (list of tuple of float, optional) – List of ranges to exclude specified as a list of (frequency_min, frequency_max).

• drag (float, optional) – Overrides the drag coefficient to this particular value.

• fixed_diode (float, optional) – Fix diode frequency to a particular frequency.

• fixed_alpha (float, optional) – Fix diode relaxation factor to particular value.

Raises:

• ValueError – If physical parameters are provided that are outside their sensible range.

• ValueError – If the distance from the bead center to the surface is set smaller than the bead radius.

• ValueError – If the hydrodynamically correct model is enabled, but the distance to the surface is specified below 0.75 times the bead diameter (this model is not valid so close to the surface).

• NotImplementedError – If the hydrodynamically correct model is selected in conjunction with axial force calibration.

• RuntimeError – If active calibration is selected, but the driving peak can’t be found near the guess of its frequency.

References

[1]

Berg-Sørensen, K. & Flyvbjerg, H. Power spectrum analysis for optical tweezers. Rev. Sci. Instrum. 75, 594 (2004).

[2]

Tolić-Nørrelykke, I. M., Berg-Sørensen, K. & Flyvbjerg, H. MatLab program for precision calibration of optical tweezers. Comput. Phys. Commun. 159, 225–240 (2004).

[3]

Hansen, P. M., Tolic-Nørrelykke, I. M., Flyvbjerg, H. & Berg-Sørensen, K. tweezercalib 2.1: Faster version of MatLab package for precise calibration of optical tweezers. Comput. Phys. Commun. 175, 572–573 (2006).

[4]

Berg-Sørensen, K., Peterman, E. J. G., Weber, T., Schmidt, C. F. & Flyvbjerg, H. Power spectrum analysis for optical tweezers. II: Laser wavelength dependence of parasitic filtering, and how to achieve high bandwidth. Rev. Sci. Instrum. 77, 063106 (2006).

[5]

Tolić-Nørrelykke, S. F, and Flyvbjerg, H, “Power spectrum analysis with least-squares fitting: amplitude bias and its elimination, with application to optical tweezers and atomic force microscope cantilevers.” Review of Scientific Instruments 81.7 (2010)

[6]

Tolić-Nørrelykke S. F, Schäffer E, Howard J, Pavone F. S, Jülicher F and Flyvbjerg, H. Calibration of optical tweezers with positional detection in the back focal plane, Review of scientific instruments 77, 103101 (2006).

Examples

import lumicks.pylake as lk
f = lk.File("calibration_file.h5")
force_slice = f.force1x

# Decalibrate existing data
volts = force_slice / force_slice.calibration[0]["Response (pN/V)"]

# Determine calibration factors using the viscosity of water at T=25 C.
lk.calibrate_force(volts.data, bead_diameter=0.58, temperature=25, sample_rate=78125)

# Determine calibration factors using the hydrodynamically correct model
lk.calibrate_force(
    volts.data,
    bead_diameter=4.89,
    temperature=25,
    sample_rate=78125,
    hydrodynamically_correct=True
)

# Determine calibration factors using the hydrodynamically correct model with active
# calibration at a distance of 7 micron from the surface
driving_data = f["Nanostage position"]["X"]
lk.calibrate_force(
    volts.data,
    bead_diameter=4.89,
    temperature=25,
    sample_rate=78125,
    hydrodynamically_correct=True,
    distance_to_surface=7,
    driving_data=driving_data.data,
    driving_frequency_guess=37,
    active_calibration=True
)