Ultrastructural Expansion Microscopy in Three In Vitro Life Cycle Stages of <i>Trypanosoma cruzi </i>

Mar&#237;a Azul de Hern&#225;ndez; Gonzalo Martinez Peralta; Rodrigo Vena; Victoria Lucia Alonso

doi:10.3791/65381

Ultrastructural Expansion Microscopy in Three In Vitro Life Cycle Stages of Trypanosoma cruzi ...

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Ultrastructural Expansion Microscopy in Three In Vitro Life Cycle Stages of Trypanosoma cruzi

Ultrastructural Expansion Microscopy in Three In Vitro Life Cycle Stages of Trypanosoma cruzi

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09:45 min

May 12, 2023

DOI: 10.3791/65381-v

María Azul de Hernández*¹, Gonzalo Martinez Peralta*^1,2, Rodrigo Vena^1,4, Victoria Lucia Alonso^1,3,4

¹Instituto de Biología Molecular y Celular de Rosario (IBR),CONICET, ²Area Biología, Facultad de Ciencias Bioquímicas y Farmacéuticas,Universidad Nacional de Rosario (UNR), ³Area Parasitología, Facultad de Ciencias Bioquímicas y Farmacéuticas,Universidad Nacional de Rosario (UNR), ⁴Unidad de Microscopía, Instituto de Biología Molecular y Celular de Rosario (IBR),CONICET

Overview

This study presents a protocol for ultrastructure expansion microscopy applied to the three in vitro lifecycle stages of Trypanosoma cruzi, the causative agent of Chagas disease. It offers a method that enhances imaging capabilities, allowing for detailed three-dimensional reconstructions of cellular structures.

Key Study Components

Research Area

Cell biology
Microscopy
Pathogen research

Background

Trypanosoma cruzi lifecycle stages
Chagas disease pathology
Importance of high-resolution imaging techniques

Methods Used

Ultrastructure expansion microscopy
Trypanosoma cruzi as the biological system
Fluorescent labeling and confocal microscopy

Main Results

Successful imaging of nanoscale structures using expansion microscopy
High-resolution insights into cell cycle phases and differentiation of T. cruzi
Potential integration with super-resolution techniques for enhanced detail

Conclusions

This study demonstrates a novel microscopy technique to advance T. cruzi research.
The findings contribute to our understanding of Chagas disease and provide methods for future cellular imaging studies.

Frequently Asked Questions

What is ultrastructure expansion microscopy?

It is a technique that allows for the visualization of fine cellular structures by expanding the sample, enabling high-resolution imaging with conventional microscopes.

Why is T. cruzi significant in this study?

T. cruzi is the pathogen responsible for Chagas disease, making it critical for understanding its biology and pathogenic mechanisms.

What are the advantages of this microscopy method?

It is compatible with standard microscopes, making it more accessible for many laboratories compared to traditional super-resolution and electron microscopy.

What do the findings contribute to the field?

They provide insights into the cellular architecture and lifecycle of T. cruzi, aiding in the understanding of its biology and potential treatment targets.

How can this method benefit other areas of research?

This technique can be applied to other cell types and organisms, facilitating advancements in cell biology, developmental biology, and immunology.

What cellular structures can be examined using this approach?

The method allows for detailed examination of cytoskeletal proteins and other cellular components at nanoscale resolution.

Can this technique be combined with other imaging methods?

Yes, this protocol suggests integration with super-resolution techniques for enhanced imaging beyond the conventional limits.

This study shows a detailed protocol to perform ultrastructure expansion microscopy in three in vitro life cycle stages of Trypanosoma cruzi, the pathogen responsible for Chagas disease. We include the optimized technique for cytoskeletal proteins and pan-proteome labeling.

This protocol details the method of ultrastructure expansion microscopy in three in vitro lifecycle stages of Trypanosoma cruzi, the pathogen responsible for Chagas disease. This protocol offers an alternative to the current super resolution imaging and electron microscopy techniques, with the advantage of being compatible with conventional microscopes found in most biology labs and in machine core facilities. It uses common techniques for most research labs to obtain images that enables three-dimensional reconstruction.

The protocol facilitates the study of nanoscale structures in trypanosomatids allowing the inspection of a population of cells and imaging the entire volume of a cell of interest at a high resolution. It is particularly useful for science-specific cell types in transient phases of the cell cycle and differentiation. To gain even more resolution, we will use the combination of ExM with super resolution microscopic techniques in order to reach resolutions below 20 nanometers.

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