Optical Photothermal Infrared-Fluorescence In Situ Hybridization (OPTIR-FISH)

Zhongyue Guo; Yeran Bai; F&#225;tima C. Pereira; Ji-Xin Cheng

doi:10.3791/66562

JoVE Journal
Bioengineering

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JoVE Journal Bioengineering

Optical Photothermal Infrared-Fluorescence In Situ Hybridization (OPTIR-FISH)

광학 광열 적외선-형광 현장 혼성화(OPTIR-FISH)

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04:07 min

February 23, 2024

DOI: 10.3791/66562-v

Zhongyue Guo*¹, Yeran Bai*^2,3, Fátima C. Pereira⁴, Ji-Xin Cheng^1,5

¹Department of Biomedical Engineering,Boston University, ²Neuroscience Research Institute, Department of Molecular, Cellular, and Developmental Biology,University of California Santa Barbara, ³Photothermal Spectroscopy Corp., ⁴School of Biological Sciences,University of Southampton, ⁵Department of Electrical & Computer Engineering, Photonics Center,Boston University

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Overview

This article presents a protocol using optical photothermal infrared-fluorescence in situ hybridization (OPTIR-FISH) to identify individual cells and analyze their metabolism at the single-cell level. The methodology aims to provide insights into cellular interactions within complex environments.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Metabolic Analysis

Background

Understanding individual cell metabolism is crucial for revealing complex biological activities.
Current methods face challenges in differentiating cellular identity while analyzing metabolism.
OPTIR-FISH combines vibrational imaging with fluorescence in situ hybridization.
This approach is compatible with identifying microbial species.

Purpose of Study

To develop a method for identifying individual cells.
To analyze metabolism at the single-cell level.
To understand cellular interactions in native environments.

Methods Used

Optical photothermal infrared imaging.
Fluorescence in situ hybridization.
Single-cell resolution techniques.
Analysis of cellular metabolism.

Main Results

Successful identification of individual cells using OPTIR-FISH.
Insights into cellular metabolism in complex environments.
Demonstrated compatibility with existing identification methods.
Potential for broad applications in biological research.

Conclusions

OPTIR-FISH is a promising tool for studying single-cell metabolism.
It enhances understanding of cellular interactions and physiology.
This methodology could lead to significant advancements in biological research.

Frequently Asked Questions

What is OPTIR-FISH?

OPTIR-FISH is a protocol that combines optical photothermal infrared imaging with fluorescence in situ hybridization to analyze individual cell metabolism.

How does OPTIR-FISH improve upon traditional methods?

It allows for the identification of individual cells while simultaneously analyzing their metabolic processes, overcoming limitations of traditional methods.

What are the applications of this research?

The methodology can be applied broadly in various fields, including neuroscience and cell biology, to study cellular interactions and metabolism.

What challenges does this study address?

It addresses the challenge of differentiating cellular identity while elucidating metabolism at the single-cell level.

What insights can be gained from this research?

The research provides insights into the heterogeneous activities of cells within complex biological systems.

여기에서는 MIP-FISH(mid-infrared photothermal-FISH)라고도 하는 광학 광열 적외선 형광 in situ hybridization(OPTIR-FISH)을 사용하여 개별 세포를 식별하고 세포의 신진대사를 이해하는 프로토콜을 제시합니다. 이 방법론은 single-cell resolution으로 세포 대사를 매핑하는 것을 포함하여 다양한 응용 분야에 광범위하게 적용할 수 있습니다.

우리의 연구 범위는 OPTIR-FISH를 개발 및 적용하여 개별 세포를 식별하고 단일 세포 수준에서 신진대사를 분석하는 것입니다. 우리는 개별 세포가 복잡한 환경 내에서 어떻게 상호 작용하는지 이해하고 그들의 생리와 경로를 이해하는 것을 목표로 합니다. 개별 세포 수준, 특히 토착 환경 내에서 신진대사를 분석하면 생물학적 시스템의 이질적인 복잡한 활동을 밝히는 데 귀중한 통찰력을 제공할 수 있습니다.

그러나 세포 정체성을 분화하고 단일 세포 수준에서 신진대사를 규명하는 동시 작업은 여전히 어려운 과제로 남아 있습니다. 당사의 고급 진동 이미징 플랫폼인 Optical Photothermal Infrared는 단일 세포 대사에 대한 연구를 제공합니다. 동시에, 미생물 종의 식별을 위해 잘 정립된 산림 in situ hybridization과 자연적으로 매우 호환됩니다.

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