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Updated: Jul 31, 2025

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

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Targeted Enzyme Activity Imaging with Quantitative Phase Microscopy.

Swati Tanwar¹, Lintong Wu¹, Noah Zahn²

¹Department of Mechanical Engineering, Johns Hopkins University, Baltimore, Maryland 21218, USA.

|May 8, 2023

Summary

This summary is machine-generated.

Researchers developed novel nanoprobes for quantitative phase imaging (QPI) to visualize enzyme activity within cells. This breakthrough enables sensitive, label-free detection of intracellular biomolecules, advancing disease diagnosis and treatment evaluation.

Keywords:

Bio-orthogonal reactions Enzymes Quantitative phase imaging Refractive index imaging Silica nanoparticles Targeted imaging

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Area of Science:

Biomedical Optics
Nanotechnology
Molecular Imaging

Background:

Quantitative phase imaging (QPI) offers label-free, 3D monitoring of cells and tissues.
Intracellular molecular imaging, particularly for enzymes, is underdeveloped in QPI.

Purpose of the Study:

To introduce novel QPI contrast agents for sensitive intracellular biomolecule detection.
To enable high-contrast refractive index (RI) imaging of enzyme activity using bio-orthogonal nanoprobes.

Main Methods:

Designed silica nanoparticles (SiO2 NPs) with higher RI than cellular components.
Functionalized NPs with cyanobenzothiazole-cysteine (CBT-Cys) and enzyme-responsive peptides.
Utilized QPI for in situ visualization of nanoprobe aggregation and intracellular RI changes.

Main Results:

Nanoprobes specifically aggregated in cells exhibiting target enzyme activity.
Increased intracellular RI was observed, enabling precise visualization of enzyme activity.
Demonstrated a new method for sensitive, label-free intracellular enzyme detection.

Conclusions:

The developed QPI-nanoprobes allow sensitive, high-contrast imaging of intracellular enzyme activity.
This approach enables spatial-temporal mapping of enzyme activity.
Potential applications include disease diagnosis and therapeutic efficacy evaluation.