Methods, systems and apparatuses for testing and calibrating fluorescent scanners

摘要

Disclosed are calibration apparatuses for fluorescent microscopy instruments and methods of making and using them. Specifically, disclosed are calibration apparatuses with a fluorescent layer, such as photoresist, deposited on a substrate, with an optional layer of a reflective material, such as chrome. Illumination of the fluorescent and/or reflective layers, and detection and analysis of the resulting emissions allows evaluation of the instrument with respect to both reflective and fluorescent channels. Selection of appropriate fluorescent materials for the one or more fluorescent layers allows the evaluation of an instrument with respect to different fluorophores, as would be used with an instrument capable of two color detection. Inclusion of a reflective layer further allows the evaluation and calibration of all optical channels of an instrument, including the reflective channel and two or more fluorescent channels, with a single calibration apparatus for imaging criteria such as uniformity, contrast and emission signal strength.

主权项

1. A method of evaluating fluorescence detection performance by a fluorescent microscopy instrument, the method comprising:
illuminating a calibration target, wherein the calibration target comprises a substrate, a reflective layer on the substrate, an outer fluorescent layer, and an optically transparent spacing layer between the reflective layer and the outer fluorescent layer, and wherein the outer fluorescent layer is illuminated with light configured to produce fluorescent emissions; detecting the fluorescent emissions with a fluorescent microscopy instrument; and analyzing the fluorescent emissions with a computer, wherein the computer comprises a computer software program, and wherein analyzing comprises comparing the fluorescent emissions to one or more pre-established criteria.