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.
  Giovedì 15/09   09:00 - 13:00   Aula E - Rosalind Franklin   V - Biofisica e fisica medica   Presentazione
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.