Fluorescence imaging autofocus systems and methods

摘要

Quantitative fluorescence imaging systems and methods using angular illumination to obtain automatic focus information. Laser scanning (e.g., point or line scanning) with angular illumination in combination with an area imaging sensor, such as with a bi-telcentric scanner, is used to determine sample height (relative to a detection axis orthogonal to a platform holding the sample) and also correct for sample height in subsequent scans.

主权项

1. A fluorescence imaging system, comprising:
a sample platform for holding a fluorescent material; a light detector having an array of sensing locations for detecting light emitted from the fluorescent material; an optical imaging system positioned between the sample platform and the light detector and configured to focus light emitted from field points on the sample platform onto the light detector, wherein contiguous field points on the sample platform are simultaneously imaged onto contiguous sensing locations on the light detector; an illumination system including a light source that emits excitation light in an absorption band of the fluorescent material, wherein the illumination system provides a beam of illumination that impinges on the fluorescent material at an angle relative to a detection axis, wherein a plane of incidence of the beam includes the detection axis and a scan direction, wherein the scan direction is perpendicular to the detection axis; a scanning mechanism that enables continuous scanning of the beam of illumination relative to the sample platform along the scan direction; and an intelligence module coupled to the light detector and configured to determine variations along the scan direction of a height of the fluorescent material parallel to the detection axis based on variations along the scan direction of positions on the light detector of maximum illuminance detected by the light detector.