PHOTOTHERMAL DEFLECTION SPECTROSCOPY OF PHOTOSYNTHETIC PIGMENTS in vivo and in vitro

Abstract— This Technical Note describes the design of a photothermal beam deflection apparatus which allows the easy and rapid measurement of thermal dissipation of absorbed light energy in various photosynthetic materials including whole plant leaves. This system is based on the "mirage effect" in which the refractive index gradient induced in a fluid in contact with the sample, irradiated with an intensity-modulated light, causes the periodic deflection of a laser beam parallel to the sample surface. The deflection of the probe laser beam is detected by a position sensor, the output of which is processed by a lock-in amplifier. Photothermal deflection signals can be monitored in vivo in intact leaves placed in various (liquid or gaseous) environments with a satisfactory signal-to-noise ratio between 100 (in water) and 50 (in air) at low modulation frequencies (ca 30 Hz). It is shown that this new and simple photothermal technique is a very sensitive tool for the measurement of absorption spectra of photosynthetic pigments both in vivo (leaves, algae or chloroplasts) and in model systems (Langmuir-Blodgett and solid films of chlorophyll).