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Samantha Gunn Mayes¹, Craig Browning^1,2, Samuel A Mayes^1,2

¹Department of Chemical and Biomolecular Engineering.

Proceedings of Spie--The International Society for Optical Engineering

|May 28, 2021

Summary

This summary is machine-generated.

This study introduces a novel excitation scanning hyperspectral microscopy system. It significantly enhances signal strength and reduces acquisition time for fluorescence imaging, improving multi-label identification.

Keywords:

Bioimaging Fluorescence HSI Imaging Microscope Microscopy Spectral Spectroscopy

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Area of Science:

Microscopy
Spectroscopy
Optical Engineering

Background:

Fluorescence imaging microscopy offers high specificity via labeling and optical filtering.
Spectral imaging enables quantitative identification of multiple fluorescent labels.
Traditional hyperspectral imaging fluorescence microscopy has long acquisition times.

Purpose of the Study:

To develop a new spectral imaging approach sampling the fluorescence excitation spectrum.
To increase signal strength for enhanced spectral, spatial, and temporal sampling.
To overcome temporal limitations of traditional hyperspectral imaging.

Main Methods:

Developed a novel excitation scanning hyperspectral microscopy system.
Utilized solid-state LEDs (365-425 nm) and a 2-mirror assembly.
Performed prototyping, benchtop testing, optical characterization, and data collection.

Main Results:

Achieved over a 10-fold increase in signal-to-noise ratio compared to emission scanning.
Demonstrated a novel configuration to reduce long image acquisition times.
Successfully assembled and characterized a system with 12 LEDs, 12 lenses, and mirrors.

Conclusions:

The new excitation scanning system offers enhanced signal strength and sensitivity.
This approach may significantly reduce image acquisition times in hyperspectral imaging.
Enables high specificity and sensitivity for real-time multi-label and autofluorescence imaging.