Fluorescence lifetime imaging microscopy method having time-correlated single-photon counting, which method permits higher light intensities

US10520434B2 · kind B2 · utility

Assignee

• LEICA MICROSYSTEMS CMS GMBH · DE

Inventors

• Volker Seyfried · Schallstadt, DE
• Bernd Widzgowski · Dossenheim, DE
• Frank Hecht · Weimar, DE

Key dates

• Filing date: May 24, 2017
• Grant date: Dec 31, 2019
• Expiry date: May 24, 2037

Classification

• Technology area (CPC G): Physics
• CPC primary: G01N21/6458
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

A fluorescence lifetime imaging microscopy method with time-correlated single photon counting includes periodically exciting a sample to emit fluorescence photons, with a measurement interval being defined between each two successive excitation light pulses. A value characterizing fluorescence decay behavior is determined based on detection times of detected fluorescence photons, and imaging is performed based one the value. An analog detector signal is sampled within a plurality of sampling intervals within a respective one of the measurement intervals and is converted into a sequence of discrete signal values associated with the sampling intervals. It is determined based thereon whether more than a predefined number of fluorescence photons has been detected within the respective measurement interval. If more than the predefined number of fluorescence photons has been detected, the respective measurement interval is discarded in the step of determining the value characterizing the fluorescence decay behavior.