Fondazione Istituto Italiano Di Tecnologia

Baylor College of Medicine

Politecnico Di Milano

Universitá Degli Studi Di Genova

Paolo Bianchini

Alberto Diaspro

Giuseppe Vicidomini

Federica Villa

Peter Saggau

Alberto Tosi

Marco Castello

Takahiro Deguchi

Mauro Buttafava

Colin J R Sheppard

Luca Lanzano'

Giorgio Tortarolo

Simonluca Piazza

A method for operating a point laser-scanning microscope includes scanning a sample with a focused illumination laser beam; recording a plurality of images by detecting elements being configurable to an intensity mode, in which the recorded images are intensity images g(n) related to photons collected during an entire dwell time of the illumination beam on an individual position n, or to a time-resolved mode, in which the recorded images are time-resolved images g(n, t), the collected photons being discriminated based on their arrival times to individual detecting elements; calculating a fingerprint image a by summing the plurality of intensity images g(n) over all positions n; estimating shift matrices sand sfrom the intensity images g(n); reconstructing at least one of a time-resolved object function fand an intensity object function f; and visualizing at least one of a high-resolution time-resolved image fand a high-resolution intensity image f.

Time-resolved imaging method with high spatial resolution

Method for increasing the optical resolution of a stimulated emission depletion microscope, or STED microscope (Stimulated Emission Depletion), based on the modulation of the intensity of a STED beam on an arbitrary time scale during the acquisition of an image and the analysis of the induced dynamics, without increasing the intensity of the STED beam and in a simple and economic manner.

Method of stimulated emission depletion microscopy having high spatial resolution

Lightening method of at least one biological sample (S), in which said at least one biological sample includes at least one or more fluorophores, at the focal point (F) of at least one objective lens (L) having a main optical axis (z-z), the method comprising the following operational steps:&#x2014;lightening (step) said at least one biological sample (S) with at least one excitation beam (EB), which propagates between said at least one objective lens (L) and said at least one biological sample (S) along at least one first propagation axis (a-a);&#x2014;lightening (step) said at least one biological sample (S) with at least two depletion beams (DB, DB'), which propagate between said at least one objective lens (L) and said at least one biological sample (S) along the respective second propagation axes (b-b, said depletion beams being donut-shaped, each one in a plane orthogonal to the respective second propagation axis (b-b, b&#x2032;-b&#x2032;); whereby said at least one first propagation axis (a-a) and said at least second propagation axes (b-b, b&#x2032;-b&#x2032;) are angularly inclined with each other, and said at least one first propagation axis (a-a) and said second propagation axes (b-b, b&#x2032;-b&#x2032;) intersect on said at least one biological sample (s) only at the focal point (F) of said at least one objective lens (L), so that an effective fluorescence volume (FV) is generated in said at least one biological sample (S) which is limited both orthogonally and axially with respect to said main optical axis (z-z).

Method of lightening at least one biological sample, three-dimensional high resolution depletion microscopy method and corresponding microscope

Optical scanning system, comprising an optical system for guiding a first and a second light beam, and deflector devices for deflecting first and second light beams in a directionally variable manner. The deflector devices comprise at least one acousto-optic deflector, and the optical system is arranged in such a way that the first and second light beams are counter-propagating through the acousto-optic deflector, which is controllable for deflecting the first and second light beams simultaneously or in pulse sequence. STED microscopy apparatus comprising an optical scanning system based on acousto-optic deflectors.

Random access stimulated emission depletion (STED) microscopy

Growing community of inventors

Genoa, Italy

Paolo Bianchini