User:Pranav Rathi/Notebook/OT/2012/11/13/Optical Tweezers Calibration
To perform accurate measurements with optical tweezers, it is necessary to calibrate some experimental parameters and even before that study the active components of the optical tweezers setup. The parameters I want to calibrate: Stiffness; which measures the force that is exerts on the trapped bead, and Sensitivity; which measures the relative position of bead within the trap. The components I want to study: Laser, AOM (acoustooptic module), microscope objective, z and x piezo, lens and mirrors, QPD (quadrant photo diode), and electronic filters. It is necessary to characterize these components first because not only they affect the experimental parameters but also reduce the chances to see something unexpected in data later on.
I characterized most of the active components time to time in the setup. The characterization also helped me in designing this tweezers.
I use two laser: 1064nm 2W ND:YAG crystalaser for tweezing ans 633nm 2mW He-Ne for surface detection and alignment purposes. It was important to know some facts about IR laser. I did not do a rigorous study (because it was not necessary), just specified few parameters like power output, beam waist and its location, polarization, beam mode profile and beam propagation factor. I used this information to design the tweezers expansion optics. The laser specification are given in the following link:
Acoustooptic modulator is used to modulate the laser intensity in the trap. So it is second most important component of the tweezers. AOM has two components: AOM driver and AOM module, I characterized both. The specifications are given in the link:
- AOM driver
- AOM module
I use 1st order diffraction beam from AOM to feed the tweezers. NI-DAQ controlled by feedback96_main_mx labview v7.1 program controls the AOM through analog input voltage to AOM driver. So there is a relationship between the analog input voltage and output laser power in 1st order diffraction beam. Unfortunately this relationship is not linear; it is some odd function (characteristic curve; see the second link). Once I know the curve I can calculate the laser power in the trap at particular input voltage. This information is very useful while calculating the stiffness from the cutoff frequency.
It is absolutely unpractical to measure the laser power in the trap before every power-spectrum data is acquired to calculate the stiffness (for stiffness I need cutoff frequency from power-spectrum and laser power in the trap at which the spectrum is acquired). So, it is done in advance: I record the laser power after the water-immersion objective for RF-input voltage of 1.3 to 4.9 in .2 volts increments. I put a water droplet on the objective and record the power with a power meter directly. I used Thorlabs sensor: model D10MM (S212A 10W) S/N 0938D08 and detector: PM100 S/N M00229006. Reflection loss at water-air interface is still less than 2%.
The Pictures shows the data and characteristic curve for AOM RF-input voltage Vs laser power after the water immersion objective. The curve looks exactly the same as AOM characteristic curve. The data is presented below; I measured the laser power for 10 times each. I use the curve and data in OT calibration program written in labview V9 to calculate the laser power in the trap from RF-input voltage. Laser power and cutoff frequency gives me the stiffness. To know cutoff frequency, I usually do power-spectrum at 1.45 volts; at this voltage power measurement error is 8%. That means calculated stiffness accuracy will not be better than 92%. I will discuss the final number later when i discuss the cutoff frequency and power-spectrum.
AOM RF-input voltage Vs laser power after the objective:
Note: the data is good until no change is made to the optical path/components downstream from the objective that may change the laser power at the objective.
Objective is another very important part of the tweezers. I am using Olympus UPLANSAPO (UIS 2) water immersion IR objective. The objective gives a maximum spot size of 760nm with a Rayleigh range of 567nm. The objective has 55% transmittance (55% of the input laser power makes through) and it has a collar for cover glass correction (spherical aberration gets worse with the depth in the sample, but i do not have to worry about it while doing power spectrum or DOG). The full details of the objective are available here:
X & Z-piezo
We use "Mad city lab" nano position systems; Nano-OP30 for x-piezo and Nano-F25HS for z-piezo. None of the stages required any calibration
Lens and mirrors
All the optics is rated for 1064nm.