Ligand Nano-cluster Arrays in a Supported Lipid Bilayer

Emmanuelle Benard; Fuwei Pi; Igor Ozerov; Anne Charrier; Kheya Sengupta

doi:10.3791/55060

Ligand Nano-cluster Arrays in a Supported Lipid Bilayer

JoVE Journal
Bioengineering

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JoVE Journal Bioengineering

Ligand Nano-cluster Arrays in a Supported Lipid Bilayer

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7,326 Views

10:34 min

April 23, 2017

DOI: 10.3791/55060-v

Emmanuelle Benard¹, Fuwei Pi^1,2, Igor Ozerov¹, Anne Charrier¹, Kheya Sengupta¹

¹Aix-Marseille Université,CNRS, UMR 7325, CINaM, ²State Key Laboratory of Food Science and Technology,School of Food Science of Jiangnan University

Overview

This article presents a protocol for fabricating protein nano-clusters in a supported lipid bilayer on glass substrates. This technique is compatible with advanced optical microscopy and aims to enhance studies on cell adhesion and migration.

Key Study Components

Area of Science

Cell Biology
Immunology
Biophysics

Background

The technique focuses on creating an array of protein nano-clusters.
It is designed for use with sensitive surface microscopy techniques.
Insights gained can influence understanding of T cell activation.
Potential applications extend to oncology and tissue engineering.

Purpose of Study

To investigate how nano-clustering of ligands affects T cell behavior.
To provide a reproducible method for studying cell interactions.
To enhance the understanding of cell biophysics.

Methods Used

Cleaning glass cover slides and observation chambers.
Depositing silica bead suspension onto the slides.
Utilizing advanced microscopy techniques like TIRF and RICM.
Implementing a straightforward protocol for reproducibility.

Main Results

The method is easily reproducible in standard biophysical labs.
Compatible with advanced microscopy for detailed analysis.
Provides insights into immunological processes.
Applicable to various cell types beyond T cells.

Conclusions

This technique offers a novel approach to studying cell adhesion.
It can significantly contribute to research in cell biology and immunology.
Future applications may extend to oncology and tissue engineering.

Frequently Asked Questions

What is the main goal of this technique?

The main goal is to fabricate protein nano-clusters in a lipid bilayer for studying cell interactions.

How does this method benefit cell biology research?

It provides insights into T cell activation and adhesion, which are crucial for understanding immune responses.

Is the protocol easy to reproduce?

Yes, the technique is designed to be easily reproducible in standard biophysical labs.

What microscopy techniques are compatible with this method?

The method is compatible with TIRF and RICM microscopy techniques.

Can this technique be applied to other cell types?

Yes, it has potential applications in various cell types beyond T cells, including oncology and tissue engineering.

We present a protocol to functionalize glass with nanometric protein patches surrounded by a fluid lipid bilayer. These substrates are compatible with advanced optical microscopy and are expected to serve as platform for cell adhesion and migration studies.

The overall goal of this technique is to fabricate an array of protein nano-clusters in a supported lipid bilayer, which is compatible with sensitive surface microscopy techniques for cell biology applications. This method can help answer key questions in the field of cell biophysics, in particular, how the nano-clustering of ligands influences T cell activation and adhesion. The main advantage of this technique is that it is easily reproducible in standard biophysical lab and it is compatible with advanced surface sensitive microscopy technique, such as TIRF and RICM.

Though this technique is designed to provide insight into immunology, it can be applied to other cell types with potential applications in cell biology, oncology and tissue engineering. To begin this procedure, clean the glass cover slides and observation chambers as outlined in the text protocol. Then, deposit 70 microliters of 2%silica bead suspension drop by drop onto a cover slide held at a 15 degree incline.

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