Solid fluorescence standard
Inventors
Riley, Patrick L. • Abbott, Richard David • Nay, Lyle M. • FOWDEN, Sarah M.
Assignees
Publication Number
US-11340169-B2
Publication Date
2022-05-24
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 relates to a solid fluorescence standard designed for calibrating and/or normalizing scientific instruments that generate and collect fluorescence data. This standard comprises an adhesive, such as a low viscosity, substantially optically transparent, solvent-free, radiation curable adhesive, with a selected quantity of fluorescent particles dispersed within it. The fluorescent particles may include quantum dots, powdered fluorescent glass, or fluorescent plastic, and are mixed with the adhesive before being cured and solidified, resulting in a robust, solid standard.
The problem addressed by this invention involves several shortcomings of conventional fluorescence standards and dyes, including poor long-term stability, susceptibility to photobleaching, limited spectral range, high cost, and mechanical, thermal, or chemical instability. Existing mineral standards can also suffer from bulk material non-uniformity and fragility, limiting their utility.
By providing a solid matrix embedding substantially non-photobleaching fluorescent particles that remain chemically stable and produce reliable, repeatable emissions over a wide spectral range, the invention overcomes these limitations. The system allows easy preparation of uniform standards across multi-well formats and different instruments, and configurations such as trimmable sheets or integration into multi-well plates provide versatility for various calibration and normalization needs.
Claims Coverage
There are two independent claims, each describing the main inventive features of a solid fluorescence standard and its preparation.
Solid matrix with evenly dispersed substantially non-photobleaching fluorescent particles
A solid fluorescence standard comprising a solid matrix and a selected quantity of substantially non-photobleaching fluorescent particles dispersed in the solid matrix. The substantially non-photobleaching fluorescent particles are one or more of quantum dots, powdered fluorescent glass, or fluorescent plastic. The solid matrix is formed from a liquid adhesive having a viscosity of 10-100 centipoise at 15-25° C. prior to curing. The fluorescent particles are evenly dispersed in the liquid adhesive before and after curing.
Solid matrix formed by curing liquid adhesive with dispersed fluorescent particles
A solid fluorescence standard comprising a solid matrix made from a cured liquid adhesive, and a selected quantity of substantially non-photobleaching fluorescent particles dispersed in the cured liquid adhesive. The fluorescent particles are evenly dispersed in the liquid adhesive before and after curing, and the uncured liquid adhesive has a viscosity of 10-100 centipoise at 15-25° C. prior to curing.
The inventive features center on a solid fluorescence standard with substantially non-photobleaching fluorescent particles uniformly dispersed in a specifically formulated adhesive matrix that is cured from a liquid state with defined viscosity.
Stated Advantages
The solid fluorescence standard provides long term stability and can be reused for weeks, months, or years without significant change in fluorescence.
The fluorescent material is chemically stable and not readily subject to photobleaching, ensuring consistent performance over time.
Applicable to a broad spectral range as fluorescent particles (e.g., quantum dots) can be selected for various absorption and emission wavelengths in the UV, visible, and IR spectra.
Uniformity is achieved within and across standards, enabling reproducible and consistent calibration of instruments, including in multi-well formats.
The standard is physically robust and can be fabricated in various formats, such as sheets or multi-well plates, and can be trimmed to fit different instruments.
Preparation is straightforward, with the ability for automated or reproducible filling in multi-well formats and minimized risk of contamination or human error.
Documented Applications
Calibrating and normalizing scientific instruments that generate and collect fluorescence data.
Use in multi-well format plates, such as 96-well plates, for plate reader calibration and checking alignment or illumination uniformity.
Integration into self-contained biological analysis devices or pouches, such as the FilmArray® system's second-stage PCR zone, for instrument calibration.
Use as trimmable sheets or plates for calibration of instruments that require specific shapes or sizes.
Monitoring thermal dynamic response and calibration in PCR instruments designed for real-time fluorescent monitoring.
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