Apparatus and method for focusing in fluorescence microscope

摘要

The presently claimed invention provides a focusing apparatus and method for a fluorescence microscope which is capable of shortening the time in focusing, increasing system throughput and avoiding from undesirable photo-bleaching. The fluorescence microscope of the present invention employs a portion of excitation light to form images of a sample to determine a focus plane for fluorescence imaging. As intensity of the portion of the excitation light is much higher than that of fluorescence light, the exposure time is highly reduced for image formation used for focusing purpose. The fluorescence microscope of the present invention enables to perform both predictive focusing and multiplex focusing.

主权项

1. A focusing apparatus for a fluorescence microscope, comprising:
an excitation light source for emitting excitation light used to excite a sample; a first beam splitter for reflecting the excitation light towards the sample via an objective lens, and transmitting fluorescence light emitted from the sample; and a focusing module located between the excitation light source and the first beam splitter, wherein the focusing module comprises a reflecting optical device used for light reflection and a focus imaging camera, wherein the reflecting optical device is located in a pathway of the excitation light between the excitation light source and the first beam splitter; wherein the sample reflects, backward scatters, or reflects and backward scatters a first portion of the excitation light towards the first beam splitter, and the first beam splitter reflects the first portion of the excitation light towards the reflecting optical device, and the reflecting optical device reflects a second portion of the excitation light towards the focus imaging camera; and wherein the second portion of the excitation light captured by the focus imaging camera forms at least one excitation light image of the sample for focusing of fluorescence imaging.