STED microscopy with pulsed excitation, continuous stimulation, and gated registration of spontaneously emitted fluorescence light

US9551658B2 · kind B2 · utility

Assignees

• Max-Planck-Gesellschaft zur Föderung der Wissenschaften e.V. · US
• Deutsches Krebsforschungszentrum · DE

Inventors

• Stefan W. Hell · Oebisfelde, DE
• Johann Engelhardt · Bad Schönborn, DE
• Matthias Reuss · Sundbyberg, SE
• Volker Westphal · Hannover, DE
• Christian Eggeling · Oettingen, DE
• Gael Moneron · Paris, FR
• Kyu Young Han · Orlando, US
• Giuseppe Vicidomini · Genova, IT
• Katrin Willig · Oettingen, DE

Key dates

• Filing date: May 22, 2013
• Grant date: Jan 24, 2017
• Expiry date: Jan 4, 2035

Classification

• Technology area (CPC G): Physics
• CPC primary: G02B21/16
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

In a STED fluorescence light microscope pulses of excitation light (3) are applied to a sample, which excite fluorescent entities contained in the sample for fluorescence, and which are focused on at least one focal area. Further, de-excitation light (12) is applied to the sample, which de-excites the excited fluorescent entities and which comprises an intensity zero point in the at least one focal area, as a continuous wave. Fluorescence light emitted by the excited fluorescent entities in the sample is registered after each pulse of the excitation light (3) and overlapping with applying the de-excitation light (13) with high temporal resolution between consecutive pulses of the excitation light (3).