Methods and apparatuses for structured illumination microscopy

US9348127B2 · kind B2 · utility

Assignee

• Carl Zeiss Microscopy GmbH · DE

Inventors

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

Key dates

• Filing date: Sep 22, 2009
• Grant date: May 24, 2016
• Expiry date: Nov 30, 2032

Classification

• Technology area (CPC G): Physics
• CPC primary: G02B21/008
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

In structured illumination microscopy, the multiple recording of images with different phase positions of the structuring requires a high stability in the optical arrangement and sample throughout the entire measuring process. Also, the structuring must be projected into the sample in a highly homogeneous manner. The current invention optimizes recording of individual images in order to achieve the best possible resolution in the result image even in problematic samples. An optimization of this kind can be carried out in different ways, for example, by determining an optimal adjustment for at least one illumination parameter or recording parameter or by pulsed illumination such that an excitation from a triplet state of the fluorescent dye to a higher triplet state is reduced, or by illuminating the sample with depletion light for depopulating a triplet state of the fluorescent dye, which reduces bleaching.