Optical Cross-Sectioning Multiphoton Microscopy Without Scanning
Yeda - code: T4-1266
Yaron Silberberg, Physics, Physics of Complex Systems
Fast cross-sectioning using multiphoton microscope. The conventionally used laser-scanning microscopy, confocal and multiphoton microscopy, although being capable of performing optical sectioning, requires a long image acquisition time, tens of milliseconds per section in current commercial systems, due to the scanning process. The field of confocal microscopy relies on the idea of point-by-point illumination of a sample and use mechanical scanning in order to collect an image. Multiphoton microscopes offer a different mechanism for optical sectioning and the need for rejecting out-of-focus scattering is practically eliminated. However, the process is efficient only when the peak intensity of the illuminating light is high. Thus there is a growing need to facilitate the multiphoton microscopy imaging of a sample by providing a novel illumination configuration and method of its operation.
Depth-resolved microscopy has been, for decades, practically synonymous with laser-scanning microscopy. The technique of the present invention provides for full-frame depth-resolved microscopy (or material processing), using an extremely simple setup as well as standard components, aiming at eliminating mechanical scanning across the sample thus making the image acquisition much faster.
Optical system for use in a multi-photon microscope.
Material processing, e.g. simultaneous depth-resolved modification of a transparent substrate by femtosecond radiation.
The present invention provides for fast imaging/processing of a sample without scanning.
The temporal profile of the pulse remains unchanged as it propagates through the sample.
Single-shot depth resolved microscopy is able to capture extremely rapid dynamics, up to the nanosecond regime.
The setup enables full-frame video-rate fluorescence lifetime imaging, simply by gating the CCD intensifier.
Enables utilization of structure illumination microscopy.
Can be used with practically any multiphoton process.
The present invention provides the ability for illuminating a region of a sample with dimensions many orders of magnitude larger than a diffraction-limited spot of the imaging lens arrangement used in the microscope. Using this method, full-frame depth-resolved microscopy can be achieved using an extremely simple setup and standard components. the proposed microscope utilizes a pulse manipulator arrangement including a temporal pulse manipulator configured to define a surface, which extends perpendicular to the optical axis of a microscope in the front focal plane of an imaging lens arrangement, and which is patterned to affect trajectories of light components of the input short pulse impinging onto different points of this surface to direct these light components along different optical paths.
This novel invention is not limited to imaging techniques in general and to microscopy in particular and can also be used for material processing.