Characterization of objective transmittance for optical tweezers.

We have measured the overall transmittance of a laser beam through an oil immersion objective as a function of the transverse size of the laser beam, using the dual-objective method. Our results show that the objective transmittance is not uniform and that its dependence on the radial beam's position can be modeled by a Gaussian function. This property affects the intensity distribution pattern in the sample region and should be taken into account in theoretical descriptions of optical tweezers. Moreover, one must consider this position dependence to determine the local laser power delivered at the sample region by the dual-objective method, especially when the beam overfills the objective's back entrance. If the transmittance is assumed to be uniform, the local power is overestimated.