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Related Experiment Video

Updated: Jun 7, 2026

Correlative Confocal and 3D Electron Microscopy of a Specific Sensory Cell

Correlative Confocal and 3D Electron Microscopy of a Specific Sensory Cell

Published on: July 19, 2015

Insight into cell-entry mechanisms of CPPs by electron microscopy.

Kärt Padari¹, Annely Lorents, Eija Jokitalo

¹Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia.

Methods in Molecular Biology (Clifton, N.J.)

|November 6, 2010

Summary

Cell-penetrating peptides (CPPs) utilize endocytosis for cellular entry, but a non-endocytic pathway emerges at low temperatures or high concentrations. Transmission electron microscopy (TEM) visualizes this mechanism using Nanogold™-labeled CPPs.

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Area of Science:

Biochemistry
Cell Biology
Nanotechnology

Background:

Cell-penetrating peptides (CPPs) are crucial for delivering macromolecules into cells.
Current understanding of CPP cellular entry mechanisms is limited, especially for non-endocytic pathways.
Fluorescence microscopy offers limited insights into CPP-cell surface interactions and membrane dynamics.

Purpose of the Study:

To investigate the non-endocytic cell entry mechanisms of cell-penetrating peptides (CPPs).
To develop and present a methodology for visualizing CPPs using electron microscopy.
To elucidate the ultrastructural details of CPP interaction with cellular structures.

Main Methods:

Covalent tagging of CPPs with a 1.4 nm gold cluster (Nanogold™).
Development of a flat-embedding protocol for cultured cells.
Transmission electron microscopy (TEM) for high-resolution ultrastructural analysis of Nanogold™-labeled CPPs.

Main Results:

The Nanogold™ labeling method is proposed as minimally disruptive to CPP uptake.
The protocol allows for the retention of cell monolayers in their in situ orientation.
TEM imaging provides high-contrast visualization of CPPs and their interaction with cellular components.

Conclusions:

Transmission electron microscopy (TEM) with Nanogold™-labeled CPPs is a powerful tool for studying non-endocytic uptake mechanisms.
This method offers detailed ultrastructural insights into CPP-cell interactions, complementing fluorescence microscopy.
The developed technique facilitates a deeper understanding of CPP translocation pathways.