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Method for high spatial resolution time-resolved imaging

Informatica Tsd EnSPAD detector array

Introduction

A microscopy imaging method able to provide a high‑resolution time-resolved and/or intensity image, fully compatible with existing confocal laser scanning microscopes. The invention is directed to a novel microscopy imaging method, able to provide a high‑resolution time-resolved and/or intensity image. The method is based on processing the images recorded by an image scanning microscope (ISM) in which the detector array is implemented using a novel SPAD (Single-Photon Avalanche Diode) array.

Technical features

The invention includes: 1) the method to obtain a high-resolution time-resolved and/or intensity image from an ISM system without demanding intensive work from the microscope user and 2) the architecture of the related microscope imaging system. Compared to imaging methods currently on the market, the invention allows for the ability to estimate separately the PSFs and the shift matrices and only successively an estimation of the specimen function makes the reconstruction method more robust. The estimation of the shift matrices using a phase correlation approach allows for sub-pixel results and fast computation. In addition, it provides technical advantages compared to known methods by allowing for fingerprint imaging (and as a result easy alignment of the system) as well as compatibility with time-resolved measurements.

Possible Applications

  • High-resolution Image-Scanning Microscopy;
  • High-resolution Fluorescence Lifetime Imaging;
  • STED and two-photon microscopy;
  • The use of the invention for imaging leads to an improvement in both the resolution and the signal-to-noise ratio, therefore the integration with existing instrumentation is highly-desirable.

Advantages

  • High-resolution imaging;
  • High Signal-to-Noise Ratio;
  • Compatibility with commercial microscopes;
  • Time-resolved measurements capability;
  • Low computational effort.