Fluorescence Lifetime Macro Imager for Biomedical Applications

Rajannya Sen; Alexander V. Zhdanov; Ciaran Devoy; Mark Tangney; Liisa M. Hirvonen; Andrei Nomerotski; Dmitri B. Papkovsky

doi:10.3791/64321

Fluorescence Lifetime Macro Imager for Biomedical Applications

JoVE Journal
Biology

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JoVE Journal Biology

Fluorescence Lifetime Macro Imager for Biomedical Applications

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06:01 min

April 7, 2023

DOI: 10.3791/64321-v

Rajannya Sen¹, Alexander V. Zhdanov¹, Ciaran Devoy², Mark Tangney², Liisa M. Hirvonen³, Andrei Nomerotski⁴, Dmitri B. Papkovsky¹

¹School of Biochemistry and Cell Biology,University College Cork, ²Cancer Research@UCC,University College Cork, ³Centre for Microscopy, Characterisation and Analysis (CMCA),The University of Western Australia, ⁴Physics Department,Brookhaven National Laboratory

Overview

This study presents a new optical imager designed for macroscopic photoluminescence lifetime imaging, particularly useful in biological samples like live animal tissue. The protocol allows for detailed mapping of phosphorescence lifetime and oxygen concentration, which is crucial for understanding various biological processes.

Key Study Components

Research Area

Biological imaging
Phosphorescence lifetime analysis
Oxygen concentration measurement

Background

Macroscopic imaging is essential for studying complex biological samples.
Existing methods may not efficiently analyze long-decaying samples.
This research aims to enhance visualization techniques in live tissues.

Methods Used

Optical imaging with time-correlated single-photon counting (TCSPC) mode
Application on live animal tissues
Use of custom-designed imaging modules and data processing software

Main Results

Effective visualization of lifetime and oxygen distribution
Ability to generate phosphorescence lifetime images with high contrast
Successful application in observing hypoxic conditions in tissue

Conclusions

The study demonstrates an innovative imaging approach for biological research.
Potential applications for diagnosing and treating tissue hypoxia-related conditions.

Frequently Asked Questions

What is the main application of the optical imager?

It is used for the macroscopic imaging of phosphorescence lifetimes and oxygen concentrations in biological tissues.

How does the imaging module operate?

It operates in TCSPC mode for accurate lifetime distribution visualization.

Can this method be used on other models apart from animal tissues?

Currently, it is limited to animal models, but there is potential for broader applications.

What are the key technological components involved?

Key components include a photomultiplier tube, LEDs, and a compact imaging module.

What biological phenomena can this method help study?

It can aid in the study of conditions like cancer, stroke, and inflammation related to tissue hypoxia.

How is data processed after imaging?

Data is processed using custom software to analyze phosphorescence decay and generate images.

What is the significance of phosphorescence lifetime imaging?

It provides insights into the dynamics of oxygen levels in biological tissues, which is crucial for various research fields.

This paper describes the use of a new, fast optical imager for the macroscopic photoluminescence lifetime imaging of long decay emitting samples. The integration, image acquisition, and analysis procedures are described, along with the preparation and characterization of the sensor materials for the imaging and the application of the imager in studying biological samples.

Our protocol provides detailed 2D mapping of phosphorescence lifetime and oxygen concentration in macroscopic objects, particularly live animal tissue and whole animals, by means of dedicated probes and sensing materials. This information is important for many areas of research. Our protocol uses an integrated and compact imaging module that operates in TCSPC mode and provides easy and accurate visualization of lifetime and oxygen distribution in complex biological samples, such as life tissue.

Take the Cricket adapter and inspect it from different sides. Remove the front side C-mount adapter to show the Photonis PP0360EF intensifier housed in the Cricket, and then put the C-mount adapter back. Remove the Cricket's front side C-mount adapter and insert the 650 plus/minus 50 nanometer emission filter.

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