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Updated: May 8, 2026

Excitation-Scanning Hyperspectral Imaging Microscopy to Efficiently Discriminate Fluorescence Signals

Excitation-Scanning Hyperspectral Imaging Microscopy to Efficiently Discriminate Fluorescence Signals

Published on: August 22, 2019

Hyperspectral fluorescence microfluidic (HFM) microscopy.

Giuseppe Di Caprio¹, Diane Schaak, Ethan Schonbrun

¹Rowland Institute at Harvard, Harvard University, 100 E. Land Boulevard, Cambridge MA, USA.

Biomedical Optics Express

|September 7, 2013

Summary

This summary is machine-generated.

We developed a hyperspectral imaging system for microfluidic cell analysis. This system accurately measures cell fluorescence spectra, resolving multiple fluorophores and cellular organelles.

Keywords:

(110.4234) Multispectral and hyperspectral imaging (170.3880) Medical and biological imaging (180.2520) Fluorescence microscopy

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Area of Science:

Biophotonics
Cellular Imaging
Microfluidics

Background:

Hyperspectral imaging provides rich spectral information crucial for cell analysis.
Existing methods may lack the resolution or efficiency for detailed cellular spectral characterization.

Purpose of the Study:

To develop and validate a novel hyperspectral imaging system for analyzing cells in microfluidic channels.
To demonstrate the system's capability in capturing detailed fluorescence emission spectra of cellular components.

Main Methods:

Utilized a single monochrome camera coupled with a linear variable bandpass filter (LVF).
Acquired bright field and hyperspectral fluorescence images of individual cells in flow.
Developed algorithms to determine peak emission wavelengths with high accuracy (<3 nm).

Main Results:

Achieved accurate peak wavelength determination (<3 nm accuracy) despite a 20 nm LVF bandwidth.
Successfully captured fluorescence spectra from individual, spatially resolved cellular organelles.
Demonstrated spectral resolution of multiple fluorophores within single cells.

Conclusions:

The developed system offers a sensitive and accurate method for hyperspectral analysis of cells in microfluidics.
This technology enables detailed investigation of cellular heterogeneity and organelle-specific fluorescence.
Potential applications include diagnostics, drug screening, and fundamental cell biology research.