scholarly journals Label-free Raman monitoring of extracellular matrix formation in three-dimensional polymeric scaffolds

2013 ◽  
Vol 10 (86) ◽  
pp. 20130464 ◽  
Author(s):  
Aliz Kunstar ◽  
Anne M. Leferink ◽  
Paul I. Okagbare ◽  
Michael D. Morris ◽  
Blake J. Roessler ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Raman Spectroscopy ◽  
Three Dimensional ◽  
Raman Microspectroscopy ◽  
Collagen Content ◽  
Cartilage Tissue ◽  
Fibre Optic ◽  
Label Free ◽  
Tissue Quality ◽  
Non Invasive

Monitoring extracellular matrix (ECM) components is one of the key methods used to determine tissue quality in three-dimensional scaffolds for regenerative medicine and clinical purposes. Raman spectroscopy can be used for non-invasive sensing of cellular and ECM biochemistry. We have investigated the use of conventional (confocal and semiconfocal) Raman microspectroscopy and fibre-optic Raman spectroscopy for in vitro monitoring of ECM formation in three-dimensional poly(ethylene oxide terephthalate)–poly(butylene terephthalate) (PEOT/PBT) scaffolds. Chondrocyte-seeded PEOT/PBT scaffolds were analysed for ECM formation by Raman microspectroscopy, biochemical analysis, histology and scanning electron microscopy. ECM deposition in these scaffolds was successfully detected by biochemical and histological analysis and by label-free non-destructive Raman microspectroscopy. In the spectra collected by the conventional Raman set-ups, the Raman bands at 937 and at 1062 cm −1 which, respectively, correspond to collagen and sulfated glycosaminoglycans could be used as Raman markers for ECM formation in scaffolds. Collagen synthesis was found to be different in single chondrocyte-seeded scaffolds when compared with microaggregate-seeded samples. Normalized band-area ratios for collagen content of single cell-seeded samples gradually decreased during a 21-day culture period, whereas collagen content of the microaggregate-seeded samples significantly increased during this period. Moreover, a fibre-optic Raman set-up allowed for the collection of Raman spectra from multiple pores inside scaffolds in parallel. These fibre-optic measurements could give a representative average of the ECM Raman signal present in tissue-engineered constructs. Results in this study provide proof-of-principle that Raman microspectroscopy is a promising non-invasive tool to monitor ECM production and remodelling in three-dimensional porous cartilage tissue-engineered constructs.

scholarly journals Label-free non-invasive quantitative measurement of lipid contents in individual microalgal cells using refractive index tomography

2017 ◽  
Author(s):  
JaeHwang Jung ◽  
Seong-Joo Hong ◽  
Han-Byeol Kim ◽  
Geon Kim ◽  
Moosung Lee ◽  
...  
Keyword(s):  
Refractive Index ◽  
Lipid Content ◽  
Three Dimensional ◽  
Biofuel Production ◽  
Chemical Extraction ◽  
Label Free ◽  
Non Invasive ◽  
Lipid Contents ◽  
Dry Cell Weight ◽  
Rapid Quantification

Microalgae are promising candidates for biofuel production due to their high lipid content. To facilitate utilization of the microalgae for biofuel, rapid quantification of the lipid contents in microalgae is necessary. However, conventional methods based on the chemical extraction of lipids require a time-consuming destructive extraction process. Here, we demonstrate label-free, non-invasive, rapid quantification of the lipid contents in individual micro-algal cells measuring the three-dimensional refractive index tomograms. We measure three-dimensional refractive index distributions within Nannochloropsis oculata cells and find that lipid droplets are identifiable in tomograms by their high refractive index. In addition, we alter N. oculata under nitrogen deficiency by measuring the volume, lipid weight, and dry cell weight of individual cells. Characterization of individual cells allows correlative analysis between the lipid content and size of individual cells.

Functional Imaging of Matrix Structure of Cryopreserved Engineered Tissues Using Back-Directional Gated Mesoscopic Imaging

2012 ◽  
Author(s):  
Young L. Kim ◽  
Zhengbin Xu ◽  
Altug Ozcelikkale ◽  
Bumsoo Han
Keyword(s):  
Tissue Engineering ◽  
Extracellular Matrix ◽  
Three Dimensional ◽  
Clinical Use ◽  
Matrix Structure ◽  
Non Invasive ◽  
Engineered Tissues ◽  
Cell Tissue ◽  
Matrix Interactions ◽  
Non Destructive

Successful cryopreservation of engineered tissues (ETs) can greatly advance the access and availability of cell/tissue engineering products for clinical use. One of the key challenges in cryopreserving ETs is that the functionality of ETs should be maintained throughout the preservation process. Many of the functionalities are associated with the extracellular matrix (ECM) microstructure, which in turn can be a crucial marker for the post-thaw functionality. Recent studies also reported that the ECM microstructure can be affected by freezing-induced cell-fluid-matrix interactions.1–3 Thus, it is critical to assess three-dimensional (3-D) matrix structure of cryopreserved ETs in a non-destructive, non-invasive, and rapid manner.

Effects of Mechanical Vibration on Matrix Production and Proliferation of Three-Dimensional Cultured Chondrocytes

2008 ◽  
Author(s):  
Yuta Takagi ◽  
Toshihiko Shiraishi ◽  
Shin Morishita ◽  
Ryohei Takeuchi ◽  
Tomoyuki Saito ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Signaling Pathways ◽  
Weight Bearing ◽  
Mechanical Vibration ◽  
Three Dimensional ◽  
Cartilage Tissue ◽  
Vibration Treatment ◽  
Matrix Production

This paper describes the effects of vibration stimulation on chondrocytes in three-dimensional culture in relation to the production of regenerative cartilage tissue, using collagen artificial skin as a carrier and supplementation with hyaluronic acid (used in the conservative treatment of osteoarthritis), and the mechanism of the adaptive response of chondrocytes to mechanical loading. The experimental condition imitates an environment of articular cartilage in vivo that chondrocytes are completely surrounded by the extracellular matrix and receives mechanical stimulation for the weight-bearing mechanics. Chondrocytes were isolated from articular cartilage of porcine metatarsophalangeal joints. Experiments were performed under four different culture conditions: control condition, in which chondrocytes were cultured with atelocollagen gel and collagen artificial skins, and no vibration (HA−Vib−); HA−Vib+, in which chondrocytes were cultured in atelocollagen gel and collagen artificial skins with vibration treatment for 2 weeks; HA+Vib−, in which chondrocytes were cultured in medium containing 0.1% hyaluronic acid; and HA+Vib+, in which chondrocytes were cultured in medium containing 0.1% hyaluronic acid with vibration treatment for 2 weeks. Histologic analysis was conducted at 14 days of culture. The proliferation of chondrocytes was obtained by counting the number of cells with a hemocytometer after 3, 7, 10, and 14 days of culture. The expression of Sox 9 and β-catenin was detected by western blotting analysis. Sox 9 has been reported of involvement in transcription of type IX collagen that binds cartilage-specific type II collagen fibrils. β-catenin plays an important role of signaling pathways of cell proliferation although the relationship between β-catenin and mechanical vibration stimulation has not been clarified yet. The obtained results are as follows. The mechanical vibration enhanced the thickness of extracellular matrix of chondrocytes in histologic section at 14 days of culture and increased the expression of Sox 9. In addition, the mechanical vibration significantly increased the number of chondrocytes after 10 days of culture and promoted the expression of β-catenin. These results show that mechanical vibration promotes the matrix production and proliferation of chondrocytes and that a part of important signaling pathways in relation to mechanical vibration stimulation and proliferation of chondrocytes has been revealed.

scholarly journals Non-contact, Label-free Monitoring of Cells and Extracellular Matrix using Raman Spectroscopy

10.3791/3977 ◽  
2012 ◽  
Author(s):  
Miriam Votteler ◽  
Daniel A. Carvajal Berrio ◽  
Marieke Pudlas ◽  
Heike Walles ◽  
Katja Schenke-Layland
Keyword(s):  
Extracellular Matrix ◽  
Raman Spectroscopy ◽  
Label Free

scholarly journals Raman spectroscopy enables phenotyping and assessment of nutrition values of plants: a review

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
William Z. Payne ◽  
Dmitry Kurouski
Keyword(s):  
Raman Spectroscopy ◽  
Abiotic Stresses ◽  
Molecular Techniques ◽  
Plant Diseases ◽  
Label Free ◽  
Breeding Programs ◽  
Non Invasive ◽  
Nutrition Value ◽  
Non Destructive ◽  
Selection Of

AbstractOur civilization has to enhance food production to feed world’s expected population of 9.7 billion by 2050. These food demands can be met by implementation of innovative technologies in agriculture. This transformative agricultural concept, also known as digital farming, aims to maximize the crop yield without an increase in the field footprint while simultaneously minimizing environmental impact of farming. There is a growing body of evidence that Raman spectroscopy, a non-invasive, non-destructive, and laser-based analytical approach, can be used to: (i) detect plant diseases, (ii) abiotic stresses, and (iii) enable label-free phenotyping and digital selection of plants in breeding programs. In this review, we critically discuss the most recent reports on the use of Raman spectroscopy for confirmatory identification of plant species and their varieties, as well as Raman-based analysis of the nutrition value of seeds. We show that high selectivity and specificity of Raman makes this technique ideal for optical surveillance of fields, which can be used to improve agriculture around the world. We also discuss potential advances in synergetic use of RS and already established imaging and molecular techniques. This combinatorial approach can be used to reduce associated time and cost, as well as enhance the accuracy of diagnostics of biotic and abiotic stresses.

scholarly journals Raman Imaging and Fluorescence Lifetime Imaging Microscopy for Diagnosis of Cancer State and Metabolic Monitoring

Cancers ◽  
2021 ◽  
Vol 13 (22) ◽  
pp. 5682
Author(s):  
Lucas Becker ◽  
Nicole Janssen ◽  
Shannon L. Layland ◽  
Thomas E. Mürdter ◽  
Anne T. Nies ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Fluorescence Lifetime ◽  
Drug Efficacy ◽  
Raman Microspectroscopy ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Non Invasive

Hurdles for effective tumor therapy are delayed detection and limited effectiveness of systemic drug therapies by patient-specific multidrug resistance. Non-invasive bioimaging tools such as fluorescence lifetime imaging microscopy (FLIM) and Raman-microspectroscopy have evolved over the last decade, providing the potential to be translated into clinics for early-stage disease detection, in vitro drug screening, and drug efficacy studies in personalized medicine. Accessing tissue- and cell-specific spectral signatures, Raman microspectroscopy has emerged as a diagnostic tool to identify precancerous lesions, cancer stages, or cell malignancy. In vivo Raman measurements have been enabled by recent technological advances in Raman endoscopy and signal-enhancing setups such as coherent anti-stokes Raman spectroscopy or surface-enhanced Raman spectroscopy. FLIM enables in situ investigations of metabolic processes such as glycolysis, oxidative stress, or mitochondrial activity by using the autofluorescence of co-enzymes NADH and FAD, which are associated with intrinsic proteins as a direct measure of tumor metabolism, cell death stages and drug efficacy. The combination of non-invasive and molecular-sensitive in situ techniques and advanced 3D tumor models such as patient-derived organoids or microtumors allows the recapitulation of tumor physiology and metabolism in vitro and facilitates the screening for patient-individualized drug treatment options.

Surface-enhanced spatially-offset Raman spectroscopy (SESORS) for detection of neurochemicals through the skull at physiologically relevant concentrations

The Analyst ◽  
2020 ◽  
Vol 145 (5) ◽  
pp. 1885-1893 ◽  
Author(s):  
Amber S. Moody ◽  
Taylor D. Payne ◽  
Brian A. Barth ◽  
Bhavya Sharma
Keyword(s):  
Raman Spectroscopy ◽  
Neurological Disease ◽  
Label Free ◽  
Detection Techniques ◽  
Non Invasive ◽  
Surface Enhanced

Detection techniques for neurotransmitters that are rapid, label-free, and non-invasive are needed to move towards earlier diagnosis of neurological disease.

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