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Multimodal Optical Imaging Platform for Studying Cellular Metabolism

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SRpHi ratiometric pH biosensors for super-resolution microscopy.

Douglas S Richardson^1,2, Carola Gregor³, Franziska R Winter³

¹Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, 37077, Göttingen, Germany. drichardson@fas.harvard.edu.

Nature Communications

|September 20, 2017

Summary

Researchers developed new ratiometric pH sensors for STED nanoscopy to study endosomal acidification in live cells. These advanced fluorescent probes enable nanoscale visualization of vesicle dynamics during endocytosis.

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Area of Science:

Cell Biology
Microscopy
Biochemistry

Background:

Fluorescence biosensors enable real-time monitoring of cellular processes.
Intracellular fluorescent pH probes are crucial for live-cell microscopy.
Endosomal acidification during endocytosis is vital for cellular functions but difficult to study at the nanoscale.

Purpose of the Study:

To develop novel ratiometric pH probes for live-cell STED nanoscopy.
To enable nanoscale investigation of endosomal pH dynamics during endocytosis.

Main Methods:

Design and synthesis of a family of ratiometric endosomal pH probes.
Optimization of probe delivery to endosomes in live cells.
Application of the probes in STED (Stimulated Emission Depletion) super-resolution microscopy.

Main Results:

Successful development of ratiometric pH sensors tailored for STED nanoscopy.
Demonstrated utility of the probes for visualizing nanoscale endosomal dynamics.
Optimized probe delivery enhances their effectiveness in live-cell imaging.

Conclusions:

The developed ratiometric pH probes are effective tools for nanoscale pH monitoring in endosomes.
These probes advance the study of endocytosis and other vesicle-mediated processes at the resolution limit of light microscopy.