Correlative AFM with Fluorescence Spectroscopy

Correlate biomolecules activity observed by Fluorescence Spectroscopy (FCS, PIE-FRET and FLIM) with the associated sample topology and mechanics observed by AFM.

Our Confocal-AFM correlative setup is mounted on top of a Zeiss inverted microscope. Fast AFM imaging (1 frame in few seconds) can be performed simultaneously with the confocal fluorescence microscopy operation. Two avalanche photodetectors (APDs) ensure two colors imaging and spectroscopy capabilities. Designed in a Pulsed Interleaved Excitation (PIE-FRET) operational scheme.

• X - Y scan (Tip scanning): 100 micron each
• Z scan: 15 micron (Head) or 10 micron (Sample, TAO stage)
• Equipped with the ultra-High-Speed cantilever holder that can achieve a scan speed of 1 image/second, a petri dish heater for living cells experiments at 37 °C and with the Advanced Quantitative-Imaging (QI) option.

• Supercontinuum laser (Leukos) that permits to select any visibile excitation laser wavelength
• Oil immersion objective from Zeiss, 1.4 NA, 100X
• Two APDs: a PicoQuant and a PerkinHelmer for two colors imaging and spectroscopy.