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Updated: May 19, 2026
Longitudinal Morphological and Physiological Monitoring of Three-dimensional Tumor Spheroids Using Optical Coherence Tomography
Published on: February 9, 2019
The use of spectroscopic imaging and mapping techniques in the characterisation and study of DLD-1 cell spheroid
Jenny Z Zhang1, Nicole S Bryce, Rainer Siegele
1School of Chemistry, The University of Sydney, NSW, 2006, Australia.
Abstract:
Determining the chemical and biological compositions of the tumour models used in pharmacological studies is crucial for understanding the interactions between the drug molecules and the tumour micro-environment. Conventional techniques for spheroid characterisation require intensive chemical pre-treatments that result in the removal of unbound metabolites. In this study, the spectroscopic techniques, scanning transmission ion microscopy (STIM), proton-induced X-ray emission (PIXE) mapping, scanning X-ray fluorescence microscopy (SXFM), and Fourier transform infrared (FT-IR) imaging were employed to gain complementary information on the compositions of untreated DLD-l cancer cell spheroids. When used together, these techniques exhibited great potential for providing a comprehensive over-view of the density, biochemistry and elemental compositions within the different regions of the spheroids. STIM density and elemental maps correlated well with cellular density across the spheroid, and showed the accumulation of S, Cu and various lighter elements in the necrotic region. High levels of oxidative stress were evident in the hypoxic region, and different degrees of cellular necrosis as well as high levels of lactate and collagen within the necrotic region were suggested by FT-IR markers. FT-IR imaging was further employed to study the pharmacodynamics of known the cytotoxins, cisplatin and Pt1C3. Cisplatin was observed to induce minimal biochemical changes to the spheroids following 24 hour incubations, whereas Pt1C3 caused severe cellular damage to the spheroid periphery; consistent with their different modes of action.
Insights
This study used advanced spectroscopy to analyze untreated cancer spheroids, revealing detailed chemical and biological compositions. These methods offer a comprehensive view of tumor microenvironments for better drug development.
Area of Science:
- Biophysics
- Chemical Biology
- Cancer Research
Background:
- Characterizing tumor models is vital for drug development.
- Conventional methods remove crucial unbound metabolites.
Purpose of the Study:
- To comprehensively analyze untreated DLD-1 cancer cell spheroids using complementary spectroscopic techniques.
- To investigate the pharmacodynamics of cytotoxins cisplatin and Pt1C3 on cancer spheroids.
Main Methods:
- Scanning transmission ion microscopy (STIM) for density mapping.
- Proton-induced X-ray emission (PIXE) and scanning X-ray fluorescence microscopy (SXFM) for elemental mapping.
- Fourier transform infrared (FT-IR) imaging for biochemical analysis.
Main Results:
- STIM revealed correlations between cellular density and elemental distribution, noting S and Cu accumulation in necrotic regions.
- FT-IR identified oxidative stress in hypoxic regions and indicated necrosis, lactate, and collagen in necrotic areas.
- Pt1C3 induced significant peripheral damage, while cisplatin showed minimal biochemical changes in spheroids.
Conclusions:
- Combined spectroscopic techniques provide a detailed overview of spheroid composition and regional variations.
- These methods are valuable for understanding drug interactions within the tumor microenvironment.
- The study demonstrated differential responses of spheroids to cisplatin and Pt1C3, highlighting their distinct mechanisms of action.
