Interference Total Internal Reflection Microscopy Particle 2D-tracking with high spatial and temporal resolution x

Experimental Setup 50mm 200mm Laser YAG (532 nm) BS 50/50 M1 100mm PlanApo 60x NA=1.45 coverslip bead M2 M3 160mm 200mm 50mm BS 95/5 camera M4 PhotoDiode 120cm 4cm AOM  /2  4MHz  =1e e+14Hz e+14Hz 

PlanApo 60x NA=1.45 coverslip bead 50mm Interference of 2 colored beams

ITIRM-Progressive waves x z coverslip v  =2  4MHz =197nm  =200nm The light diffused by the bead is modulated at high frequency  with the amplitude related to the position along the z axis and the phase proportional to the position along the x axis

Synchronous Detection AOM Photo Diode 90deg LowPass BW=300kHz

x t Phase variation = movement along X APD AOM  (x)=2rad A(z)=1.0V

Phase variation = movement along X x t APD AOM  (x)=4rad A(z)=1.0V

Drift ~ 1 nm/s Phase-Position Calibration

Single 10 nm step RMS noise ± 2 Å Piezo: 10 nanometers steps Testing ITIRM with a piezo BW=300kHz

z t Amplitude variation = movement along Z APD AOM  (x)=2rad A(z)=1.0V

z t Amplitude variation = movement along Z APD AOM A(z)=0.5V  (x)=2rad

Time resolution ~ 2 µs RMS noise ~ 2 Å Drift ~ 1 nm/s ITIRM-Summary