Ultrasensitive bioassay platform with an ultra-large dynamic range using microlaser ensemble quenching
NIBIB - National Institute of Biomedical Imaging and Bioengineering
About This Grant
SUMMARY Analog enzyme-linked immunosorbent assay (ELISA) is a commonly used technique, in which the detection signal varies continuously with analyte concentration. To improve the detection limit, digital ELISA was developed, which allocates individual analytes to an ensemble of microunits (such as microbeads) and then counts the fraction of the “bright microunits” that emit light. While very sensitive, digital ELISA has a very limited dynamic range due to the fundamental assumption it relies on, i.e., the averaged analytes per microunit is far below 1. To extend the dynamic range, samples need to be serially diluted to a concentration within the digital ELISA dynamic range. However, the appropriate dilution factor needs to be determined through multiple trials. For multiplexed detection involving multiple analytes with vastly different concentrations, it is impossible to find a one-size-fits-all dilution factor. Other strategies include stitching the digital and analog calibration curves or extrapolating the digital calibration curve beyond the single-molecule assumption. However, it is difficult to determine the cut-off concentration between digital and analog mode and there is a discontinuity in digital and analog calibration curves due to two completely different methods used to obtain the corresponding detection signals. All these lead to large measurement errors. Here we propose a microlaser ensemble quenching bioassay platform that achieves an ultra-high sensitivity and ultra-large dynamic range with a unified method and without artificial digital-to-analog stitching. A microlaser ensemble consists of thousands of microfabricated high quality vertical cavity surface emitting lasers (VCSELs), which has a lasing threshold distribution when the microlaser ensemble is exposed to analytes. By scanning the VCSEL pumping level and counting the fraction of the bright VCSELs, we essentially probe the lasing threshold distribution, which in turn maps the analyte distribution in the microlaser ensemble. Similar to digital ELISA, the analyte distribution in an ensemble relates to analyte concentration in solution, which can be established through a statistical model. However, in contrast to digital ELISA that cannot differentiate a microunit with 1 analyte from that with more than 1 analyte, our method takes advantage of the non-linear (or threshold) behavior of laser emission and the tunable pumping level to turn the VCSELs on and off to differentiate the VCSELs with different analytes (i.e., mapping the distribution), thus significantly increasing the dynamic range. There are two specific aims. Aim 1. Fabricate and characterize VCSEL ensembles. We will fabricate arrays of microfluidic VCSELs using semiconductor microfabrication technologies. Each array will consist of 10,000 microfluidic VCSELs. The VCSELs’ quality and the lasing threshold distribution will be characterized in the absence and presence of quenchers. Aim 2. Develop an assay protocol and test the assay platform. We will use interleukin-6 in buffer and in serum as a model system. IL- 6 concentration will be varied from 0.01 pg/mL to 106 pg/mL to cover a range of eight orders of magnitude. The detection variability, detection limit, dynamic range, and recovery rate will be characterized.
Grant Summary
Ultrasensitive bioassay platform with an ultra-large dynamic range using microlaser ensemble quenching is a NIBIB - National Institute of Biomedical Imaging and Bioengineering grant providing up to $429K for university, nonprofit, healthcare org. Applications are due 2028-03-01 (open). Check eligibility and apply with FindGrants.
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Up to $429K
2028-03-01
- 1Confirm your organization is eligible for Ultrasensitive bioassay platform with an ultra-large dynamic range using microlaser ensemble quenching from NIBIB - National Institute of Biomedical Imaging and Bioengineering, checking organization type, location, and any population or project requirements.
- 2Gather the required documents and information, including your organization details, project plan, and budget figures.
- 3Draft your application narrative and budget addressing the funder's priorities and review criteria. FindGrants can draft each section for you to review and edit.
- 4Review every section against the requirements checklist, then export a submission-ready application pack and submit it to NIBIB - National Institute of Biomedical Imaging and Bioengineering before the deadline.
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Ultrasensitive bioassay platform with an ultra-large dynamic range using microlaser ensemble quenching: Frequently Asked Questions
Who is eligible for the Ultrasensitive bioassay platform with an ultra-large dynamic range using microlaser ensemble quenching?
Ultrasensitive bioassay platform with an ultra-large dynamic range using microlaser ensemble quenching is offered by NIBIB - National Institute of Biomedical Imaging and Bioengineering and is generally open to university, nonprofit, healthcare org. It is open to organizations nationwide unless the funder specifies otherwise. Review the specific eligibility terms before applying, since funders set their own requirements around organization type, location, and the population or project being served.
How much funding does the Ultrasensitive bioassay platform with an ultra-large dynamic range using microlaser ensemble quenching provide?
Ultrasensitive bioassay platform with an ultra-large dynamic range using microlaser ensemble quenching provides up to $429K per award from NIBIB - National Institute of Biomedical Imaging and Bioengineering. Actual award sizes depend on the scope of your project, available program funds, and the number of applicants, so build a budget that reflects realistic, allowable costs rather than the maximum figure.
When is the Ultrasensitive bioassay platform with an ultra-large dynamic range using microlaser ensemble quenching deadline?
Applications for Ultrasensitive bioassay platform with an ultra-large dynamic range using microlaser ensemble quenching are due 2028-03-01 (open). Because deadlines can change, verify the date with the funder, NIBIB - National Institute of Biomedical Imaging and Bioengineering, and give yourself enough time to prepare a complete, competitive application before the close date.
How do you apply for the Ultrasensitive bioassay platform with an ultra-large dynamic range using microlaser ensemble quenching?
To apply for Ultrasensitive bioassay platform with an ultra-large dynamic range using microlaser ensemble quenching, confirm your eligibility, gather the required documents, and prepare a narrative and budget that address the funder's priorities. FindGrants guides you step by step and can draft each section, then exports a submission-ready application pack for this grant from NIBIB - National Institute of Biomedical Imaging and Bioengineering.