Excitation–emission matrix fluorescence spectroscopy for cell viability testing in UV-treated cell culture

Głowacz, Klaudia; Skorupska, Sandra; Grabowska-Jadach, Ilona; Ciosek-Skibińska, Patrycja

doi:10.1039/d1ra09021f

Cited by 8 publications

(17 citation statements)

References 29 publications

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“…Based on the analysis of the presented spectra, one can see that the exposure of A375 and HaCaT cells to oxaliplatin results in the most significant alternations of the fluorescence signals around λ ex ∈ (250 nm, 300 nm), λ em ∈ (300 nm, 450 nm), which might correspond to the changes of the content of such endogenous fluorophores as aromatic amino acids. ,, Moreover, the difference spectra reveal that in the mentioned spectral range, a substantial increase in the fluorescence can be observed, and its decrease at approximately 290 nm/350 nm, which suggests the presence of at least two fluorophores (Figure C,F). No significant changes in the spectral regions most typically attributed to the content of cofactors and vitamins , were observed between control samples and samples of cell cultures affected by the highest concentration of oxaliplatin (Figure A,B and D,E) compared to the previously reported results . On the other hand, based on the difference EEM spectra, it can be seen that fluorescence intensity in the ranges of λ ex ∈ (300 nm, 340 nm), λ em ∈ (360 nm, 460 nm), and λ ex ∈ (340 nm, 430 nm), λ em ∈ (420 nm, 480 nm) slightly decreases under the influence of the highest concentration of oxaliplatin.…”

Section: Resultscontrasting

confidence: 46%

Excitation–Emission Matrix Fluorescence Spectroscopy Coupled with PARAFAC Modeling for Viability Prediction of Cells

et al. 2023

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Cell-based sensors and assays have great potential in bioanalysis, drug discovery screening, and biochemical mechanisms research. The cell viability tests should be fast, safe, reliable, and time- and cost-effective. Although methods stated as “gold standards”, such as MTT, XTT, and LDH assays, usually fulfill these assumptions, they also show some limitations. They can be time-consuming, labor-intensive, and prone to errors and interference. Moreover, they do not enable the observation of the cell viability changes in real-time, continuously, and nondestructively. Therefore, we propose an alternative method of viability testing: native excitation–emission matrix fluorescence spectroscopy coupled with parallel factor analysis (PARAFAC), which is especially advantageous for cell monitoring due to its noninvasiveness and nondestructiveness and because there is no need for labeling and sample preparation. We demonstrate that our approach provides accurate results with even better sensitivity than the standard MTT test. With PARAFAC, it is possible to study the mechanism of the observed cell viability changes, which can be directly linked to increasing/decreasing fluorophores in the cell culture medium. The resulting parameters of the PARAFAC model are also helpful in establishing a reliable regression model for accurate and precise determination of the viability in A375 and HaCaT-adherent cell cultures treated with oxaliplatin.

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Section: Resultscontrasting

confidence: 46%

Excitation–Emission Matrix Fluorescence Spectroscopy Coupled with PARAFAC Modeling for Viability Prediction of Cells

et al. 2023

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“…[72][73][74][75] Therefore, considering the materials, the set of 3D printing approaches should have acceptable cell viability if living cells are printed. For the fabrication procedures, UV crosslinking period, [77][78][79][80] crosslinking/liquefaction temperatures [81] in PBE, and immerging periods in CaCl 2 and sodium citrate baths [71] all have controllable effects on cell viability [82,83] if cell-laden bioinks are utilized, which will be further investigated in the future to alleviate cell damage during fabrication. However, if cells are seeded on the crosslinked patch or brain model, the negative effects of both materials and fabrication procedures can be neglected.…”

Section: Conflict Of Interest Statementmentioning

confidence: 99%

3D printing‐based full‐scale human brain for diverse applications

Hua

Zhang

Raymond

et al. 2023

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Surgery is the most frequent treatment for patients with brain tumors. The construction of full-scale human brain models, which is still challenging to realize via current manufacturing techniques, can effectively train surgeons before brain tumor surgeries. This paper aims to develop a set of three-dimensional (3D) printing approaches to fabricate customized full-scale human brain models for surgery training as well as specialized brain patches for wound healing after surgery. First, a brain patch designed to fit a wound's shape and size can be easily printed in and collected from a stimuli-responsive yield-stress support bath. Then, an inverse 3D printing strategy, called "peeling-boiled-eggs," is proposed to fabricate full-scale human brain models. In this strategy, the contour layer of a brain model is printed using a sacrificial ink to envelop the target brain core within a photocurable yield-stress support bath. After crosslinking the contour layer, the as-printed model can be harvested from the bath to photo crosslink the brain core, which can be eventually released by liquefying the contour layer. Both the brain patch and fullscale human brain model are successfully printed to mimic the scenario of wound healing after removing a brain tumor, validating the effectiveness of the proposed 3D printing approaches. K E Y W O R D S3D printing, brain patch, full-scale human brain models, stimuli-responsive yield-stress bath, surgery training, wound healing | INTRODUCTIONPrimary malignant brain tumors, such as glioblastoma multiforme, [1,2] are the tenth leading cause of death for human beings. In 2020, approximately 85,000 people were diagnosed with brain tumors, and more than 700,000 people had to live with a primary malignant brain tumor in the United States. [3][4][5] Although treatment for brain tumors is affected by various factors, including type, location and size of a tumor, patient's age and health, and so on, surgery is still the most This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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“…Another powerful tool for analyzing viability in bioprocess monitoring is fluorescence spectroscopy since living cells contain multiple endogenous fluorophores which are mainly involved in metabolic activity or cellular growth. Amongst these fluorophores, fluorescent amino acids (phenylalanine, tyrosine and tryptophan), cofactors (FAD, FMN, NADH and NADPH) porphyrins and vitamins (riboflavin) are the most significant ones when it comes to determining the viability of biological samples via fluorescence spectroscopy [ 2 , 45 , 46 ]. Based on the existence or absence and the amount of these substances, fluorescence spectroscopy offers a wide range of information on the cellular state, and thus, has been used for bioprocess monitoring for many years [ 47 , 48 ].…”

Section: Spectroscopy-based Techniques For On-line Monitoring Of Viab...mentioning

confidence: 99%

“…Multivariate data analysis is required to extract relevant information from fluorescence spectra. Since the information on cell viability included in the fluorescence fingerprint is not readily accessible, deconvolution can be accomplished using chemometric methods [ 45 , 49 , 50 ].…”

Section: Spectroscopy-based Techniques For On-line Monitoring Of Viab...mentioning

confidence: 99%

“…The results were compared to MTT test data as a reference method and unfolded partial least squares (UPLS) regression was applied to verify whether the changes in EEMs could be related with cell viability. The proposed method achieved a high determination coefficient of R 2 = 0.986, and thus, provides a promising basis for the development of a spectroscopic soft sensor [ 45 ]. The application of such a spectrofluorometric soft sensor in monitoring the viability (among other parameters) of industrially relevant CHO cells has been investigated in multiple research projects up to a bioreactor volume of 5000 L [ 49 , 54 , 55 , 56 , 57 ].…”

Section: Spectroscopy-based Techniques For On-line Monitoring Of Viab...mentioning

confidence: 99%

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Sensors and Techniques for On-Line Determination of Cell Viability in Bioprocess Monitoring

et al. 2022

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In recent years, the bioprocessing industry has experienced significant growth and is increasingly emerging as an important economic sector. Here, efficient process management and constant control of cellular growth are essential. Good product quality and yield can only be guaranteed with high cell density and high viability. Whereas the on-line measurement of physical and chemical process parameters has been common practice for many years, the on-line determination of viability remains a challenge and few commercial on-line measurement methods have been developed to date for determining viability in industrial bioprocesses. Thus, numerous studies have recently been conducted to develop sensors for on-line viability estimation, especially in the field of optical spectroscopic sensors, which will be the focus of this review. Spectroscopic sensors are versatile, on-line and mostly non-invasive. Especially in combination with bioinformatic data analysis, they offer great potential for industrial application. Known as soft sensors, they usually enable simultaneous estimation of multiple biological variables besides viability to be obtained from the same set of measurement data. However, the majority of the presented sensors are still in the research stage, and only a few are already commercially available.

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Excitation–emission matrix fluorescence spectroscopy for cell viability testing in UV-treated cell culture

Abstract: Excitation-emission matrix fluorescence spectroscopy can be applied for label-free and non-destructive determination of cells viability, which is promising methodology for drug screening, biocompatibility testing, or pharmacodynamic studies.

Cited by 8 publications

References 29 publications

Excitation–Emission Matrix Fluorescence Spectroscopy Coupled with PARAFAC Modeling for Viability Prediction of Cells

Excitation–Emission Matrix Fluorescence Spectroscopy Coupled with PARAFAC Modeling for Viability Prediction of Cells

3D printing‐based full‐scale human brain for diverse applications

Sensors and Techniques for On-Line Determination of Cell Viability in Bioprocess Monitoring

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