Increased depth-resolution microscopy

US9201011B2 · kind B2 · utility

Assignee

• Carl Zeiss Microscopy GmbH · DE

Inventors

• Thomas Kalkbrenner · Jena, DE
• Helmut Lippert · Jena, DE
• Ingo Kleppe · Jena, DE

Key dates

• Filing date: Nov 11, 2011
• Grant date: Dec 1, 2015
• Expiry date: Nov 11, 2031

Classification

• Technology area (CPC G): Physics
• CPC primary: G01N21/64
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

A method for high-resolution luminescence microscopy of a sample marked with marking molecules that can be activated to excite particular luminescent radiation, including: repeated activation of a subset of the marking molecules to emit luminescent radiation; repeated imaging of the sample along a depth direction and with a predetermined optical resolution; and producing images from the repeated imaging. Locations of the marking molecules are determined with a spatial resolution that is increased above the predetermined optical resolution. Activation of the marking molecules can be through radiation introduced into multiple regions, each extending along a plane substantially perpendicular to the depth direction. The regions can be arranged so that the regions are behind one another and overlap only partially. Separate images of the sample may be recorded for activation in each of the regions in order to obtain depth information relating to the marking molecules from the separate images.