Combination microscopy

US8704196B2 · kind B2 · utility

Assignee

• Carl Zeiss Microscopy GmbH · DE

Inventors

• Ralf Wolleschensky · Jena, DE
• Ingo Kleppe · Jena, DE
• Gerhard Krampert · Jena, DE
• Michael Kempe · Jena, DE

Key dates

• Filing date: Oct 28, 2009
• Grant date: Apr 22, 2014
• Expiry date: Feb 14, 2030

Classification

• Technology area (CPC G): Physics
• CPC primary: G02B27/58
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

A method for generating an image of a sample by a microscopy method including varying local resolution, wherein at least two of the following microscopy methods are combined: laser scanning microscopy, a microscopy method wherein the sample is excited to luminescence by structured line or wide area illumination, and a first microscopy image is generated from the images thus obtained, having increased local resolution greater than the optical resolution of the image, a further microscopy method according to the PAL principle, by which a second microscopy image is generated, indicating geometric locations of marker molecules emitting luminescent radiation at an increased local resolution relative to the optical resolution, and a further microscopy method, wherein the sample is marked using marking molecules suitable for the STED, ESA, or RESOLFT technique, and a third microscopy image is generated of STED, ESA, or RESOLFT, wherein the obtained images are superimposed.