Multi-confocal fluorescence correlation spectroscopy in living cells using a complementary metal oxide semiconductor-single photon avalanche diode array

M. Kloster-Landsberg (1) , D. Tyndall (2) , I. Wang (1) , R. Walker (2) , J. Richardson (2) , R. Henderson (2) , A. Delon (1)

1 MOTIV-LIPhy - Matériaux, Optique et Techniques Instrumentales pour le Vivant
2 Institute for Integrated Micro and Nano Systems

Living cells are heterogeneous and rapidly changing biological samples. It is thus desirable to measure molecular concentration and dynamics in many locations at the same time. In this note, we present a multi-confocal setup capable of performing simultaneous fluorescence correlation spectroscopy measurements, by focusing the spots with a spatial light modulator and acquiring data with a monolithic 32 × 32 single-photon avalanche photodiode array. A post-processing method is proposed to correct cross-talk effects between neighboring spots. We demonstrate the applicability of our system by simultaneously measuring the diffusion of free enhanced Green Fluorescent Protein (eGFP) molecules at nine different points in living cells.

Domaines: Biophysique [physics.bio-ph]; Biologie cellulaire

M. Kloster-Landsberg, D. Tyndall, I. Wang, R. Walker, J. Richardson, et al.. Multi-confocal fluorescence correlation spectroscopy in living cells using a complementary metal oxide semiconductor-single photon avalanche diode array. Review of Scientific Instruments, 2013, 84 (7), pp.076105. ⟨10.1063/1.4816156⟩. ⟨hal-00958200⟩