Method for evaluating fluorescence results in a microscope image

US8988771B2 · kind B2 · utility

Assignee

• Carl Zeiss Microscopy GmbH · DE

Inventors

• Ingo Kleppe · Jena, DE
• Thomas Kalkbrenner · Jena, DE
• Ralf Wolleschensky · Jena, DE

Key dates

• Filing date: Jun 19, 2010
• Grant date: Mar 24, 2015
• Expiry date: Feb 24, 2031

Classification

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

Abstract

The invention allows a quantitative evaluation of images acquired by microscope having fewer errors and is applicable in connection with high-resolution methods, particular at a high speed. A microscope image is analyzed in which the intensity distributions of fluorescence events have in each instance a diffraction-dependent extent which corresponds to an extent of a point spread function of the microscope and are arranged so as to be spatially non-overlapping, or at least predominantly spatially non-overlapping, in that at least one counter is initialized for every region to be analyzed in the microscope image, at least one fluorescence event is identified in a region to be analyzed in the microscope image, and the counter corresponding to the relevant region is incremented for each fluorescence event identified in the region. The counting results in a dramatic improvement in the signal-to-noise ratio at a high evaluation speed.