Dear community,
On a Zeiss LSM 980, we’re trying to start FCS measurements along multiple points on the Z axis. The FCS measurement itself runs fine. We thought we’d first set the ideal z position to zero, then start macro recording where we
- move the z position (e.g. 10 microns)
- measure FCS
and repeat these two steps (ideally set as a loop but not absolutely necessary).
The issues we’re having:
- Any change in z position (e.g. an increment of 1.0 microns in the “focus” tab) records both absolute and relative numbers in the macro
- when removing the absolute numbers but retaining the relative ones, the focus moves accordingly in z (as judged on the touchscreen panel) and stays there for a second or so, but then moves back to the original position to run the measurement.
The scope is an upright 980 and software is Zen Blue (I think 3.10; the macro records as Zen26).
A sample macro text is below but as said it repeats the measurement at z=0.
Ideally it would also fit the FCS data and possibly export the data automatically, but those are future problems.
If I may, I’m tagging @sebi06 here
hardwaresetting1 = ZenHardwareSetting()
hardwaresetting1.SetParameter('MTBLKM980Laser561', 'IsEnabled', 'true')
Zen.Devices.ApplyHardwareSetting(hardwaresetting1)
hardwaresetting1.SetParameter('MTBLSM880DetectorMeta', 'DetectorGain', '1045')
Zen.Devices.ApplyHardwareSetting(hardwaresetting1)
hardwaresetting1.SetParameter('MTBLSM880DetectorLong', 'DetectorGain', '990')
Zen.Devices.ApplyHardwareSetting(hardwaresetting1)
experiment1 = Zen.Acquisition.Experiments.GetByName("fcs.czexp")
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = 10.0
Zen.Devices.Focus.Apply()
fcsimage1 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = 9.0
Zen.Devices.Focus.Apply()
fcsimage2 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = 8.0
Zen.Devices.Focus.Apply()
fcsimage3 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = 7.0
Zen.Devices.Focus.Apply()
fcsimage4 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = 6.0
Zen.Devices.Focus.Apply()
fcsimage5 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = 5.0
Zen.Devices.Focus.Apply()
fcsimage6 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = 4.0
Zen.Devices.Focus.Apply()
fcsimage7 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = 3.0
Zen.Devices.Focus.Apply()
fcsimage8 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = 2.0
Zen.Devices.Focus.Apply()
fcsimage9 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = 1.0
Zen.Devices.Focus.Apply()
fcsimage10 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = 0.0
Zen.Devices.Focus.Apply()
fcsimage11 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = -1.0
Zen.Devices.Focus.Apply()
fcsimage12 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = -2.0
Zen.Devices.Focus.Apply()
fcsimage13 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = -3.0
Zen.Devices.Focus.Apply()
fcsimage14 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = -4.0
Zen.Devices.Focus.Apply()
fcsimage15 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = -5.0
Zen.Devices.Focus.Apply()
fcsimage16 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = -6.0
Zen.Devices.Focus.Apply()
fcsimage17 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = -7.0
Zen.Devices.Focus.Apply()
fcsimage18 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = -8.0
Zen.Devices.Focus.Apply()
fcsimage19 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = -9.0
Zen.Devices.Focus.Apply()
fcsimage20 = Zen.Acquisition.AcquireImage(experiment1)
# CAUTION! Risk of Crushing when executing code!
Zen.Devices.Focus.TargetPosition = -10.0
Zen.Devices.Focus.Apply()
fcsimage21 = Zen.Acquisition.AcquireImage(experiment1)