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Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Updated: Sep 10, 2025

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

Published on: February 4, 2016

Voltage-Programmed Sequential Fluorescence Encoding (VPSFE) Enables Multiplexed In Situ Proteo-Imaging with

Chen Wang¹, Jiayi Zheng¹, Zhenghan Xiong¹

¹Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education, School of Pharmacy, China Phar-maceutical University, Nanjing, 210009, China.

Angewandte Chemie (International Ed. in English)

|August 22, 2025

Summary

We developed an electrophoresis-based method for rapid, highly specific in situ imaging of proteins. This technique eliminates washing steps, enabling faster and more versatile multiplexed imaging for biological research.

Keywords:

In situ imaging Multiplexed protein imaging Single cell Turing Pattern Washing free

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Last Updated: Sep 10, 2025

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

Published on: February 4, 2016

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Area of Science:

Molecular Biology
Biotechnology
Cell Biology

Background:

DNA barcode-based immunolabeling advanced single-cell protein profiling.
Conventional multiplexed imaging suffers from slow, laborious probe removal and hybridization cycles, limiting throughput and universality.

Purpose of the Study:

To introduce a novel electrophoresis-based in situ probe removal method for high-throughput, specific multiplexed imaging.
To develop voltage-programmed sequential fluorescence encoding (VPSFE) for protein detection.
To establish a VPSFE-based Turing pattern coding strategy for multiplexed target identification.

Main Methods:

Utilized low-voltage electrophoresis for excess probe removal (2 min) and high-voltage electrophoresis for hybridized probe dissociation (3 min).
Validated robustness through 19 rounds of cyclic electrophoresis and 10 rounds of repetitive imaging.
Developed VPSFE for sequential color coding and a Turing pattern strategy for multiplexed detection using electric fields.

Main Results:

Achieved high-specificity iterative probe imaging without washing steps.
Demonstrated exceptional specificity and efficiency across 19 electrophoresis cycles and 14 probes.
Successfully applied VPSFE to image epithelial-mesenchymal transition (EMT)-related proteins.
Generated 27 unique fluorescence Turing patterns to encode 27 distinct targets using three voltage conditions and three fluorescence channels.

Conclusions:

The electrophoresis-based method significantly enhances specificity and efficiency in multiplexed in situ imaging.
VPSFE offers a rapid, universal, and probe exchange-free approach for complex biological sample analysis.
The electric-field Turing pattern coding strategy provides a novel platform for high-density multiplexed detection.