Fluorescence Imaging Autofocus Systems And Methods

Abstract

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-telecentric 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.

Description

technical field [0001] The present disclosure relates to quantitative fluorescence imaging, and more particularly to autofocus systems and methods using angular illumination. Background technique [0002] A recent improvement in fluorescence imaging is the bi-telecentric widefield fluorescence scanner, which enables accurate quantitative measurements with focus-independent pass-to-pass registration. U.S. Patent Application S / N14 / 312,409, entitled "Telecentric, Wide-Field Fluorescence Scanning Systems and Methods," filed June 23, 2014, which is incorporated herein by reference for all purposes, discusses dual Features of the telecentric wide-field fluorescence scanning system. One feature of this design is angular illumination (excitation), which has the benefit of reduced optical background and thus higher sensitivity. However, a disadvantage of corner illumination is that as the height of the sample changes, so does the position of the imaged line on the sensor. Therefor...

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Problems solved by technology

This further adds complexity and slows down the process

[0006] Furthermore, Olson's technique does not provide a detected signal that exits the detector due to changes in the z-height of the sample

Therefore, the idea of ​​obtaining images at different heights does not work because, at many z-heights, there will be no signal for detection (leaving the detector) and therefore any contrast-based scheme will not be able to detect optimal focus where are you

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