Development of a 3D oxygen measurement system for cell research
Pöppönen, Pasi (2023)
Pöppönen, Pasi
2023
Bioteknologian ja biolääketieteen tekniikan maisteriohjelma - Master's Programme in Biotechnology and Biomedical Engineering
Lääketieteen ja terveysteknologian tiedekunta - Faculty of Medicine and Health Technology
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Hyväksymispäivämäärä
2023-11-17
Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi:tuni-202310259116
https://urn.fi/URN:NBN:fi:tuni-202310259116
Tiivistelmä
Cell research is transitioning from planar 2D cell cultures towards 3D culture models. 3D cell culture models enable the monitoring of biological processes that are challenging to replicate in planar models, such as cell migration, tumor invasion and angiogenesis. The concentration of O2 within tissues and cell cultures has a crucial role in both physiological and pathological cell processes. The monitoring of O2 concentration within cell cultures is important for sustaining physiologically relevant culture environment as well as for monitoring cell metabolia.
This thesis describes and characterizes a novel measurement system that is intended for pO2 measurements of 3D cell cultures. The system combines a cell-friendly Selective Plane Illumination Microscopy (SPIM) technology with Fluorescence Lifetime Imaging (FLIM) capability. The FLIM feature is used to measure the luminescense lifetime of O2-sensitive photoluminescent sensor particles embedded in 3D cell cultures. The spatially measured luminescense lifetime is affiliated with a specific O2 by using a modified Stern-Volmer model.
The system is capable of imaging pO2 in 3D cell samples ranging from ambient to anoxic O2 conditions. The acquired calibration curve could predict pO2 at plusminus 0.98 kPa accuracy on 95% confidence interval. An optimal set of system parameters for best measurement accuracy were presented as the result of the system characterization. Thesis opens new opportunities for future studies and system development in the field of O2 monitoring applied to cell research.
This thesis describes and characterizes a novel measurement system that is intended for pO2 measurements of 3D cell cultures. The system combines a cell-friendly Selective Plane Illumination Microscopy (SPIM) technology with Fluorescence Lifetime Imaging (FLIM) capability. The FLIM feature is used to measure the luminescense lifetime of O2-sensitive photoluminescent sensor particles embedded in 3D cell cultures. The spatially measured luminescense lifetime is affiliated with a specific O2 by using a modified Stern-Volmer model.
The system is capable of imaging pO2 in 3D cell samples ranging from ambient to anoxic O2 conditions. The acquired calibration curve could predict pO2 at plusminus 0.98 kPa accuracy on 95% confidence interval. An optimal set of system parameters for best measurement accuracy were presented as the result of the system characterization. Thesis opens new opportunities for future studies and system development in the field of O2 monitoring applied to cell research.