A novel technique to control and visualize functions of living cells

Overview

A research group composed of Professor Takayuki Shibata and his colleagues at Department of Mechanical Engineering, Toyohashi University of Technology has given greater functionalities to atomic force microscopy (AFM).

Our research team has succeeded in minimally invasive surgery to living cells using photocatalytic oxidation controlled in a nanoscale space and visualizing dynamic information on intracellular biomolecules.

This proposed technique for controlling and visualizing the process of cell function expression on a high level has significant potential as a strong nanofabrication and nanomeasurement system to solve the mystery of life.

Details

An integrated understanding of life phenomena and the control thereof are absolutely essential for further development of the medical and pharmaceutical fields.

The thesis for creating life innovation is to solve the structure and function of biomolecules such as genomes, proteins, and sugar chains and also solve the function of cells, which are the basic unit for life activity.

Therefore, we aim to establish a technology for minimally invasive surgery to target living cells at a molecular level (God's hand to manipulate the function of cells) and visualizing changes in the dynamic behavior of intracellular biomolecules and the state of cell membrane protein at a single molecular level (God's eye to see the function of cells), and thus provide an innovative nanofabrication and nanomeasurement platform to solve the mystery of life.

Here, our research team has succeeded in giving two new functions to atomic force microscopy (AFM)1). The first advancement is to coat the tip apex of an AFM probe with a thin film of titanium oxide (TiO2) known as a photocatalyst.

By this method, the photocatalytic reaction is localized in a nanoscale space (100 nm region) in the vicinity of the tip apex to achieve minimally invasive cell membrane perforation.

As a result, the probability of cell membrane perforation reaches 100%, and a cell viability of 100% is also successfully achieved, allowing us to verify that minimally invasive surgery can be carried out.

The second advancement is to insert the tip apex of an AFM probe coated with silver (Ag) nanoparticles into a living cell. We have thus succeeded in acquiring a sensitive Raman spectrum originating in protein, DNA, lipids, etc. (Tip-Enhanced Raman Spectroscopy, TERS).

By this method, a difference in the ratio of biomolecules between a cell's nucleus and cytoplasm was visualized as information inside a cell, and it was found that there is an inverse correlation (a phenomenon that as one increases, the other decreases) between proteins and glycogen (also called animal starch) as temporal changes in biomolecules inside cells.

1) Atomic Force Microscopy (AFM) is a microscope that detects the atomic force affecting the tip apex and the surface of a sample and was invented by Dr. Gerd Binning and others at IBM Zurich Laboratories in 1985.

AFM is a strong tool that can directly observe atomic and molecular images and also evaluate mechanical properties such as frictional force and hardness and electric, magnetic, and thermal properties with nanoscale spatial resolution, becoming a fundamental technology leading today's nanotechnology.

Furthermore, AFM can make observations not only in the atmosphere but also in liquids, and thus has been actively applied in the life science and biotechnology fields.

Future Outline

In order to simultaneously achieve nanofabrication and nanomeasurement functions, we will establish a tip-enhanced Raman spectroscopic (TERS) function by coating the surface of a TiO2-functionalized AFM probe with Ag nanoparticles in the future.

This function will be able to visualize the process of degradation reactions of organic substances based on photocatalytic oxidation (changes in molecular structures) during the cell surgery process.

We will also aim to achieve a means for measuring a single molecule in a target cell membrane protein using the high molecular recognition ability of an antigen-antibody reaction, and we will aim to establish a technique for selective nanofabrication for a single molecule in the target membrane protein identified by the above means.

It is expected that this proposed technique could solve the mechanisms of life functions and be applied to work such as the development of novel medicines.

Source:
Journal reference:

Shibata, T., et al. (2020) Photocatalytic Nanofabrication and Intracellular Raman Imaging of Living Cells with Functionalized AFM Probes. Micromachines. doi.org/10.3390/mi11050495.

Comments

The opinions expressed here are the views of the writer and do not necessarily reflect the views and opinions of AZoLifeSciences.
Post a new comment
Post

While we only use edited and approved content for Azthena answers, it may on occasions provide incorrect responses. Please confirm any data provided with the related suppliers or authors. We do not provide medical advice, if you search for medical information you must always consult a medical professional before acting on any information provided.

Your questions, but not your email details will be shared with OpenAI and retained for 30 days in accordance with their privacy principles.

Please do not ask questions that use sensitive or confidential information.

Read the full Terms & Conditions.

You might also like...
Transforming Microbiome Research with Single-Cell Genome Analysis