Ultrahigh resolution multicolor colocalization of single fluorescent probes

US6844150B2 · kind B2 · utility

Assignee

• The Regents of the University of California · US

Inventors

• Shimon Weiss · Pinole, US
• Xavier Michalet · Paris, FR
• Thilo Lacoste · Lörrach, DE

Key dates

• Filing date: Aug 8, 2001
• Grant date: Jan 18, 2005
• Expiry date: Mar 7, 2023

Classification

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

Abstract

A novel optical ruler based on ultrahigh-resolution colocalization of single fluorescent probes is described. Two unique families of fluorophores are used, namely energy-transfer fluorescent beads and semiconductor nanocrystal (NC) quantum dots, that can be excited by a single laser wavelength but emit at different wavelengths. A novel multicolor sample-scanning confocal microscope was constructed which allows one to image each fluorescent light emitter, free of chromatic aberrations, by scanning the sample with nanometer scale steps using a piezo-scanner. The resulting spots are accurately localized by fitting them to the known shape of the excitation point-spread-function of the microscope.