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Integrative Toolkit to Analyze Cellular Signals: Forces, Motion, Morphology, and Fluorescence

Integrative Toolkit to Analyze Cellular Signals: Forces, Motion, Morphology, and Fluorescence

Published on: March 5, 2022

Rapid tool for cell nanoarchitecture integrity assessment.

Guido Gaietta¹, Mark F Swift¹, Niels Volkmann²

¹Scintillon Institute, San Diego CA 92123, USA.

Journal of Structural Biology

|September 28, 2021

Summary

A new Rapid Cell Integrity Assessment (RCIA) assay uses light microscopy to quickly screen if experimental protocols preserve nanoscale cellular structure. This accessible method ensures reliable interpretation of high-resolution cell imaging data.

Keywords:

CLEM Cryo-electron tomography In-situ cellular methods Virus inactivation

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Area of Science:

Cell Biology
Microscopy
Biophysical Techniques

Background:

High-resolution cell imaging is increasingly used, but interpreting results depends on maintaining nanoscale structural integrity.
Electron cryo-tomography is the gold standard for assessing nanoscale structural integrity but is complex, costly, and time-consuming.
There is a need for accessible methods to ensure experimental protocols do not compromise cellular nanoarchitecture.

Purpose of the Study:

To develop an accessible, inexpensive, and reliable screening assay for preserving nanoscale structural integrity in whole-cell preparations.
To validate a new assay against electron cryo-tomography for accuracy in assessing protocol compatibility.

Main Methods:

Development of the Rapid Cell Integrity Assessment (RCIA) assay.
RCIA utilizes light microscopy imaging at room temperature.
Comparison of RCIA results with cellular electron cryo-tomography as a benchmark.

Main Results:

The RCIA assay accurately identifies adverse impacts of experimental protocols on cellular nanoarchitecture.
RCIA demonstrates compatibility with assessing the effects of reagents and protocols, including those for virus inactivation or process arrest.
The assay provides a reliable screening tool for maintaining structural integrity at the nanoscale.

Conclusions:

The Rapid Cell Integrity Assessment (RCIA) assay offers an accessible and efficient method for evaluating experimental protocol compatibility with nanoscale cellular structure preservation.
RCIA serves as a valuable screening tool, complementing electron cryo-tomography by providing rapid feedback on sample integrity.
This assay facilitates more reliable interpretation of data from high-resolution cell imaging techniques.