Comunicazione
Development of a super-resolution optical microscope enabling fluorescence and label-free approaches.
Nepita I., Bianchini P., Callegari F., Castello M., Cuneo L., Sheppard C.J.R, Diaspro A.
Coupling microscopy and spectroscopy with the idea of extracting all the information carried by the collected photons may have a deep impact in biological research. We are developing a Multimodal Optical MIcroscopy Image Correlation Sensing approach, MOMIX(tm). It enables super-resolution lifetime imaging and label-free Mueller matrix polarization control and analysis. The setup is based on a laser scanning confocal microscope, which implements polarization resolved and two-photon excitation fluorescence (2PEF) imaging modalities. The first method exploits a photoelastic modulator (PEM) to modulate the polarization of the light at a resonant frequency of $50 {kHz}$. A lock-in detection scheme allows to analyse intensities differences due to alterations of the polarization state in tens of microseconds. One- and two-photon excited fluorescence can also be collected by MOMIX. In particular, fluorescence lifetime can be imaged at high spatial resolution taking advantage of the image scanning detection scheme. Real-time synchronization of electro-optical devices (scanners, PEM, detectors), pixel-by-pixel signal acquisition, and processing will be presented.