Fluorescent Calibration Target With Reflective Layer
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Solution Overview
Problem
Fluorescence microscopy instruments face challenges in accurately and efficiently calibrating fluorescent channels, particularly in creating effective fluorescent calibration targets that mimic the emission spectra and strength of biological samples, and in simultaneously testing reflective channels in a compact manner, especially for multi-color detection instruments.
Innovation Solution
The development of calibration targets with one or more fluorescent layers and optional reflective layers, deposited on a substrate, which are designed to simulate the emission characteristics of fluorescently labeled biological materials, allowing for the evaluation of fluorescence detection performance and calibration of both fluorescent and reflective channels using a computer software program to analyze detected emissions and identify calibration needs.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If multiple separate calibration targets are used for different fluorescent channels and reflective channels, then each channel can be calibrated independently, but the device complexity and calibration time increase
Solution Approach 1:
The patent combines multiple calibration functions into a single calibration target that includes both fluorescent calibration features (for fluorescent channel calibration) and reflective calibration features (for reflective channel calibration). This merging eliminates the need for multiple separate calibration targets, reducing device complexity while maintaining comprehensive calibration capabilities for all channels.
Solution Approach 2:
The calibration target is designed with multi-functionality, serving as a universal calibration tool for both fluorescent and reflective channels. The target includes diverse calibration features that can be used across different detection channels, allowing a single target to perform multiple calibration functions that previously required separate targets.
2Measurement precision
If traditional calibration targets are used that do not mimic biological sample emissions, then manufacturing is simpler, but measurement precision for fluorescent channel calibration deteriorates
Solution Approach 1:
The calibration target incorporates fluorescent materials with specific emission spectra that locally match the characteristics of biological samples. Different regions of the target can have different fluorescent properties tailored to simulate specific sample types, ensuring measurement precision while allowing flexible manufacturing approaches for different application scenarios.
3Measurement precision
If calibration procedures are manual and iterative, then adjustment precision can be high, but productivity and calibration time decrease
Solution Approach 1:
The calibration system incorporates feedback mechanisms where the instrument detects emissions from the calibration target and automatically adjusts calibration parameters based on the detected signals. This closed-loop feedback enables rapid, automated calibration iterations that maintain high precision while significantly improving productivity compared to manual calibration methods.
Solution Approach 2:
The calibration target is designed to enable self-calibration capabilities, where the instrument can automatically perform calibration by detecting the known features on the target and adjusting its parameters accordingly. This self-service calibration reduces the need for manual intervention while maintaining calibration accuracy, thereby increasing productivity.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables precise and efficient calibration of fluorescence microscopy instruments, improving the accuracy and consistency of image acquisition by identifying potential adjustments or component replacements, and facilitating the use of a single calibration target for multiple fluorescent channels and reflective channels, enhancing the overall performance of the instruments.
Implementation Method 1
illuminate the calibration apparatus, detect fluorescent emissions, and analyze the fluorescent emissions
Data Source
AI summary
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.