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Updated: Sep 24, 2025
Cellular Redox Profiling Using High-content Microscopy
Published on: May 14, 2017
High-Throughput Examination of Therapy-Induced Alterations in Redox Metabolism in Spheroid and Microtumor Models
Mans Broekgaarden1,2, Anne-Laure Bulin1,3, Tayyaba Hasan4
1Wellman Center for Photomedicine, Department of Dermatology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
Abstract:
The capacity of cancer cells to adjust their metabolism to thrive in new environments and in response to treatments has been implicated in the acquisition of treatment resistance. To optimize therapeutic strategies such as photodynamic therapy (PDT)-based combination treatments, methods to characterize the plasticity of cancer metabolism in response to treatments are required. This protocol provides a method for high-throughput and label-free tracking of metabolic redox states in cancer tissues, leveraging the autofluorescent properties of nicotinamide dinucleotide (NAD(P)H) and oxidized flavoprotein adenine dinucleotide (FAD). The methodology is optimized to be applied to 3D spheroid/microtumor/organoid cultures, regardless of the culture type (e.g., adherent or suspension cultures) and morphology. The exploitation of these methods may elucidate mechanisms of metabolic adaptation and perturbations in redox homeostasis, and chart the overall tumor health in both 3D culture models and ex vivo tissues following cancer therapies, such as PDT.
Insights
This study presents a novel method for tracking cancer cell metabolism using autofluorescence. This technique helps understand metabolic adaptation and resistance to therapies like photodynamic therapy (PDT).
Area of Science:
- Biochemistry
- Cancer Biology
- Medical Imaging
Background:
- Cancer cells exhibit metabolic plasticity, contributing to treatment resistance.
- Optimizing therapies like photodynamic therapy (PDT) requires understanding metabolic adaptation.
- Characterizing cancer metabolism in response to treatment is crucial for developing effective strategies.
Purpose of the Study:
- To develop a high-throughput, label-free method for tracking metabolic redox states in cancer tissues.
- To characterize metabolic plasticity in 3D cancer models and ex vivo tissues.
- To elucidate mechanisms of metabolic adaptation and tumor health following cancer therapies.
Main Methods:
- Utilized autofluorescence of nicotinamide dinucleotide (NAD(P)H) and oxidized flavoprotein adenine dinucleotide (FAD).
- Developed a protocol optimized for 3D spheroid, microtumor, and organoid cultures (adherent or suspension).
- Applied label-free tracking of metabolic redox states in cancer tissues.
Main Results:
- The method allows for high-throughput and label-free tracking of metabolic redox states.
- The protocol is versatile, applicable to various 3D culture types and morphologies.
- Enabled elucidation of metabolic adaptation and redox homeostasis perturbations.
Conclusions:
- This methodology provides a tool to study cancer cell metabolic plasticity in response to therapies.
- It aids in understanding tumor health and resistance mechanisms in 3D models and ex vivo tissues.
- Facilitates optimization of therapeutic strategies, including photodynamic therapy (PDT).
