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Automated System for Single Molecule Fluorescence Measurements of Surface-immobilized Biomolecules

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Structurally-thrifty and visible-absorbing fluorophores.

Xiao Luo¹, Yan Chen², Yanchun Li²

¹State Key Laboratory of Bioreactor Engineering, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China; School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200241, China.

Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy

|September 15, 2020

Summary

Researchers developed novel, small fluorophores (SDXs) using a D'D-π-A design for enhanced biological labeling. These visible-absorbing dyes show strong green fluorescence and are quenched by hydrogen-bonding solvents, enabling cellular imaging.

Keywords:

Cell imaging D′D-π-A Green emission Structurally-thrifty Visible-absorbing

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Quantification of Efferocytosis by Single-cell Fluorescence Microscopy

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Area of Science:

Organic Chemistry
Photochemistry
Biophysical Chemistry

Background:

Push-pull fluorophores are crucial for biological labeling applications.
Developing structurally minimal fluorophores with desirable photophysical properties remains a challenge.

Purpose of the Study:

To design and synthesize novel, structurally-thrifty fluorophores with visible absorption using the D'D-π-A strategy.
To investigate the photophysical properties and solvent sensitivity of the synthesized fluorophores.
To demonstrate the utility of these fluorophores for imaging cellular microenvironments.

Main Methods:

Synthesis of structurally-thrifty fluorophores (SDXs) based on the D'D-π-A design.
Spectroscopic characterization including absorption (λabs = 420 nm) and emission (λem = 530 nm) measurements.
Evaluation of fluorescence quantum yield (up to 0.84) and solvent effects (quenching in hydrogen-bonding solvents).
Cellular imaging experiments using BEAS-2B cells to demonstrate application in non-hydrogen-bonding microenvironments.

Main Results:

Successful construction of structurally-thrifty, visible-absorbing fluorophores (SDXs).
SDXs exhibit long-wavelength absorption and strong green fluorescence with high quantum yield.
Fluorescence quenching observed in hydrogen-bonding solvents (e.g., MeOH, H2O).
Demonstrated feasibility for imaging cellular non-hydrogen-bonding microenvironments in BEAS-2B cells.

Conclusions:

The D'D-π-A design strategy is effective for creating novel, structurally-thrifty fluorophores.
SDXs possess advantageous photophysical properties for biological applications.
The solvent-sensitive fluorescence of SDXs allows for specific cellular microenvironment imaging.