A novel
on-the-fly
calibration method of optical
tweezers is presented, which enables in situ control and measure of
absolute temperature and viscosity at nanoscale dimensions. Such noncontact
measurement and control at the nanoscale are challenging as the present
techniques only provide off-line measurements that do not provide
absolute values. Additionally, some of the present methods have a
low spatial resolution. We simultaneously apply the high temporal
sensitivity of position autocorrelation and equipartition theorem
to precisely measure and control in situ temperature and the corresponding
microrheological property around the focal volume of the trap at high
spatial resolution. The femtosecond optical tweezers (FOTs) use a
single-beam high repetition rate laser for optical trapping to result
in finer temperature gradients in comparison to the continuous-wave
laser tweezers. Such finer temperature gradients are due to the additional
nonlinear optical (NLO) phenomena occurring only at the nanoscale
focal plane of the FOTs. Because NLO processes are laser peak power-dependent,
they promote an effective study of physical properties occurring only
at the focal plane. Using FOTs at optically benign near-infrared wavelengths,
we demonstrate microrheological control and measurement in water by
adding a highly absorbing yet low fluorescent dye (IR780).