Solid fluorescence standard
Inventors
Riley, Patrick L. • Abbott, Richard David • Nay, Lyle M. • FOWDEN, Sarah M.
Assignees
Publication Number
US-10613031-B2
Publication Date
2020-04-07
Expiration Date
2036-12-15
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Abstract
A solid fluorescence standard that can be used to calibrate and/or normalize a device (e.g., a scientific instrument) that is configured for generating and collecting fluorescence data. A fluorescence standard disclosed herein includes an adhesive (e.g., a low viscosity, substantially optically transparent, solvent-free, radiation curable adhesive, such as, but not limited to, a UV curable adhesive), and a selected quantity of fluorescent particles (e.g., quantum dots) dispersed in the adhesive. The adhesive and the fluorescent particles are mixed together and disposed in a sample well. The adhesive is then cured and solidified, which yields a solid fluorescence standard in the well.
Core Innovation
The invention provides a solid fluorescence standard used to calibrate and/or normalize scientific instruments configured for generating and collecting fluorescence data. The key components of the fluorescence standard are an adhesive—such as a low viscosity, substantially optically transparent, solvent-free, radiation curable adhesive—and a selected quantity of substantially non-photobleaching fluorescent particles like quantum dots. The adhesive and fluorescent particles are mixed and placed into a sample well, then cured and solidified to yield a robust, solid fluorescence standard in the well.
Conventional fluorescence standards and dyes commonly suffer from deficiencies including poor long-term stability, susceptibility to photobleaching, limited spectral range, high cost, and instability from mechanical or chemical factors. Mineral-based standards, while more chemically stable, may lack uniformity and durability. The disclosed invention addresses these issues by providing fluorescence standards that remain stable and chemically robust over extended periods, and that are physically tough and uniform from standard-to-standard and across multi-well formats.
The fluorescence standard can be produced in a variety of formats, including single sample wells, multi-well plates, sheets, or pouches, with the option to use different types of fluorescent particles or mixtures to match desired emission spectra. These standards are compatible with a broad range of devices and can accommodate diverse excitation and emission wavelength requirements, making them particularly useful for calibration in various scientific and diagnostic instruments.
Claims Coverage
The patent includes several independent claims, each defining a distinct inventive feature around fluorescence standards utilizing adhesive and non-photobleaching fluorescent particles.
Fluorescence standard comprising adhesive and non-photobleaching fluorescent particles in a sample well
A fluorescence standard consists of: - An adhesive; - A selected quantity of substantially non-photobleaching fluorescent particles dispersed in the adhesive; and - A sample well containing the adhesive and particles, wherein the adhesive is cured and solidified after being disposed in the sample well.
Multi-well fluorescence standard with adhesive and non-photobleaching fluorescent particles
A multi-well fluorescence standard includes: - An adhesive; - A selected quantity of substantially non-photobleaching fluorescent particles dispersed in the adhesive; and - A multi-well format plate having multiple sample wells, at least one of which contains the adhesive and particles, with the adhesive cured and solidified after being placed in the sample well(s).
Fluorescence standard in sheet format comprised of adhesive and non-photobleaching fluorescent particles
A fluorescence standard includes: - An adhesive; - A selected quantity of substantially non-photobleaching fluorescent particles dispersed in the adhesive; - A sheet comprised of the adhesive and the fluorescent particles, with the adhesive cured and solidified after being disposed in the sheet.
Fluorescence standard with light-excluding storage container
A fluorescence standard including: - An adhesive; - A selected quantity of substantially non-photobleaching fluorescent particles dispersed in the adhesive; - A sample well having the adhesive and particles disposed and cured therein; and - A storage container for storing the standard, where the container is configured to exclude light.
These inventive features define various formats (sample well, multi-well plate, sheet, and storage container) for a robust fluorescence standard utilizing an adhesive matrix with non-photobleaching fluorescent particles, tailored for long-term calibration uses in scientific instrumentation.
Stated Advantages
The fluorescence standard exhibits long-term stability, remaining usable for weeks, months, or even years without significant degradation.
The fluorescent material is chemically stable, not readily subject to photobleaching, and provides consistent fluorescence signals over time.
The standard is applicable across a broad spectral range, as fluorescent particles can be chosen to absorb and emit at various wavelengths.
It is physically robust and uniform, enabling reproducible preparation across individual and multi-well formats.
The standard can be reused multiple times, allowing calibration of one or more instruments and facilitating data normalization over extended periods.
Documented Applications
Calibrating and/or normalizing devices or scientific instruments configured for generating and collecting fluorescence data.
Use in multi-well format plates, including 96-well plates and second-stage arrays of FilmArray pouches for real-time PCR applications.
Monitoring and calibrating the optical excitation and detection systems in instruments such as PCR thermocyclers with real-time fluorescence detection.
Tracking thermal dynamic response and temperature-related fluorescence variation in instrument sample blocks.
Providing a stable standard for comparison and normalization of instrument data collected on different days or across different instruments.
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