Enhanced Fluorophore Discrimination in Fluorescence Microscopy Using Multi View Spectral Unmixing with Photobleaching Dynamics

Background:

There are hundreds to thousands of different cells or other structures that can be labeled with fluorescent reporters in a biological sample, and then imaged on a fluorescence microscope. Unfortunately, the broad excitation and emission spectra of biologically-compatible fluorescent dyes make it difficult or impossible to reliably distinguish more than a few different fluorescent reporters in a single sample, due to fluorophore bleed-through and crosstalk. Being able to image and distinguish hundreds of fluorescent labels in a single image would be useful for spatial biology imaging, tumor cell diagnostics and biomarker profiling, microbiome imaging, and profiling. This would also have potential uses in DNA sequencing applications.

Technology Overview:

This technology significantly increases the number of fluorescent labels that can be distinguished in a single image. To do this, it combines two components. The first is a method to record spectral images of fluorescently labeled biological cells that utilizes multiple exposures of the sample to induce photobleaching. This is critical due to the fact biologically compatible fluorophores have unique bleach profiles in addition to unique spectra. The technology’s second component is an algorithm that uses multi-view machine learning to unmix the fluorescent labels, utilizing both their spectral information and their bleach characteristics as multiple views of the data.

Advantages:

The main advantage of this technology is that it can distinguish hundreds of fluorescent labels in a single image.

Applications:

•    Spatial biology imaging.
•    Tumor cell diagnostics.
•    Biomarker profiling.
•    Microbiome imaging and profiling.
•    DNA sequencing applications

Intellectual Property Summary:

Provisional Application filed 10/10/2024

Stage of Development:

TRL 3 – Experimental proof of concept

Licensing Status:

This technology is available for licensing.

Licensing Potential:

This technology would be of interest to entities involved in microbial research, including:
•    Pharmaceutical and healthcare companies.
•    Hospitals and other healthcare facilities.
•    Universities.
•    Medical research laboratories.
•    Government health organizations.