Non-invasive monitoring of functional state of articular cartilage tissue with label-free unsupervised hyperspectral imaging (Conference Presentation)

Author(s):  
Saabah B. Mahbub ◽  
Anna Guller ◽  
Martin Gosnell ◽  
Ayad G. Anwer ◽  
Ewa M. Goldys
2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Saabah B. Mahbub ◽  
Anna Guller ◽  
Jared M. Campbell ◽  
Ayad G. Anwer ◽  
Martin E. Gosnell ◽  
...  

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
C Y Tan ◽  
S B Mahbub ◽  
C A Campugan ◽  
J Campbell ◽  
A Habibalahi ◽  
...  

Abstract Study question Can we separate between control and reversine-treated cells within the inner cell mass (ICM) of the mouse preimplantation embryo by using label-free and non-invasive hyperspectral microscopy? Summary answer Hyperspectral microscopy is able to discern between control and reversine-treated cells using cellular autofluorescence in the complete absence of fluorescence tags. What is known already Embryo mosaicism (containing cells that are euploid (46 chromosomes) and aneuploid (deviation from the expected number of chromosomes)) affects up to 17.3% of human blastocyst embryos. Current diagnosis of aneuploidy in the IVF clinic involves a biopsy of trophectoderm (TE) cells or spent media followed by sequencing. In some blastocyst embryos these approaches will fail to diagnose of the proportion of aneuploid cells within the fetal lineage (ICM). Study design, size, duration The impact of aneuploidy on cellular metabolism was assessed by using cellular autofluoresence and hyperspectral microscopy (broad spectral profile). Two models were employed: (i) Primary human fibroblast cells with known karyotypes (4-6 independent replicates, euploid n = 467; aneuploid n = 969) and reversine induced aneuploidy in mouse embryos (5-8 independent replicates, 30-44 cells per group). Both models were subjected to hyperspectral imaging to quantify native cell fluorescence. Participants/materials, setting, methods The human model is comprised of euploid (male and female) and aneuploid (triploid and trisomies: 13, 18, 21, XXX, and XXY) primary human fibroblast cells. For the mouse model, we treated embryos with reversine, a reversible spindle assembly checkpoint inhibitor, during the 4- to 8-cell division. Individual blastomeres were dissociated from control and reversine treated 8-cell embryos. Blastomeres were either imaged directly or used to generate chimeric blastocysts with differing ratios of control:reversine-treated cells. Main results and the role of chance Following unsupervised linear unmixing, the relative abundance of metabolic cofactors was quantified: reduced nicotinamide adenine dinucleotide (NAD(P)H) and flavins with the subsequent calculation of the optical redox ratio (ORR: Flavins/[NAD(P)H + Flavins]). Primary human fibroblast cells displayed an increase in the relative abundance of NAD(P)H with a decrease in flavins, leading to a significant reduction in the ORR for aneuploid cells (P < 0.05). The mouse embryos displayed an identical trend as the human model between control and reversine-treated embryos. Mathematical algorithms were applied and able to distinguish between (i) euploid and aneuploid primary human fibroblast cells, (ii) control and reversine-treated mouse blastomeres and (iii) chimeric blastocysts with differing ratios of control and reversine-treated cells. The accuracy of these separations was supported by receiver operating characteristic curves with areas under the curve. We also showed that hyperspectral imaging of the preimplantation embryo does not impact on embryo developmental competence, pregnancy outcome and offspring health in a mouse model. We believe the role of chance is low as both human somatic cells and mouse embryos showed a consistent shift in cellular metabolism in response to human fibroblast cells that are aneuploid and reversine treated mouse embryos. Limitations, reasons for caution Further validation of our approach could include sequencing of the ICM of individual blastocysts to determine the proportion of aneuploid cells in ICM and correlate this with the metabolic profile obtained through hyperspectral imaging. Wider implications of the findings With hyperspectral imaging able to discriminate between (i) euploid and aneuploid human fibroblast cells and (ii) control and reversine-treated mouse embryos, this could be an accurate, non-invasive and label-free optical imaging approach to assess mosaicism within the ICM of mouse embryos, potentially leading to a new diagnostic tool for embryos. Trial registration number Not applicable


2021 ◽  
Vol 2 (43) ◽  
pp. 54-61
Author(s):  
Dmitriy A. Burynin ◽  
◽  
Aleksandr A. Smirnov

Portable spectroradiometers and hyperspectral cameras are increasingly being used to quickly assess the physiological state of plants. The operation of these devices is based on the registration of reflection or reflection and transmission spectra. (Research purpose) The research purpose is in analyzing the technical means and methods of non-invasive monitoring of the plant state based on the registration of the reflection spectra of leaves. (Materials and methods) The article presents a review of the work on the application of hyperspectral imaging methods. Authors classified and analyzed materials on spectroscopic radiometers and hyperspectral cameras, and outlined the prospects for implementation. Authors applied the methods of a systematic approach to the research problem. (Results and discussion) Hyperspectral imaging methods serve as an effective means of monitoring plants. It is possible to determine the pigment composition of plants, lack of nutrition, and detect biotic stress through hyperspectral imaging. The article presents methods of application of portable spectroradiometers and hyperspectral cameras. With the help of these devices it is possible to carry out measurements with high spectral resolution. The difficulty of accurately detecting the content of pigments in the leaves lies in the mutual overlap of the areas of light absorption by them. The main drawback of spectroradiometers is that they measure only at one point on a single sheet. The article presents the difficulties encountered in interpreting the results obtained by the hyperspectral camera. The background reflectivity of the soil, the geometry of the vegetation cover, and the uneven lighting can make errors in the measurements. (Conclusions) The article presents the disadvantages of the hyperspectral imaging method when using only the reflection spectrum. In order to increase the accuracy of the determination of pigments and stresses of various origins, it is necessary to develop a portable device that combines the methods of recording reflection and fluorescence.


2013 ◽  
Vol 10 (86) ◽  
pp. 20130464 ◽  
Author(s):  
Aliz Kunstar ◽  
Anne M. Leferink ◽  
Paul I. Okagbare ◽  
Michael D. Morris ◽  
Blake J. Roessler ◽  
...  

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.


Photonics ◽  
2021 ◽  
Vol 8 (10) ◽  
pp. 425
Author(s):  
Boris Shurygin ◽  
Olga Chivkunova ◽  
Olga Solovchenko ◽  
Alexei Solovchenko ◽  
Alexey Dorokhov ◽  
...  

We compared two approaches to non-invasive proximal sensing of the early changes in fresh-cut lettuce leaf quality: hyperspectral imaging and imaging of variable chlorophyll fluorescence contained in the leaves. The estimations made by the imaging techniques were confronted with the quality assessments made by traditional biochemical assays (i.e., relative water content and foliar pigment (chlorophyll and carotenoid) composition. The hyperspectral imaging-based approach provided the highest sensitivity to the decline of fresh-cut lettuce leaf quality taking place within 24 h from cutting. Using of the imaging pulse-amplitude modulated PAM chlorophyll fluorometer was complicated by (i) weak correlation of the spatial distribution pattern of the Qy parameter with the actual physiological condition of the plant object and (ii) its high degree of heterogeneity. Accordingly, the imaging PAM-based approach was sensitive only to the manifestations of leaf quality degradation at advanced stages of the process. Sealing the leaves in polyethylene bags slowed down the leaf quality degradation at the initial stages (<three days) but promoted its rate at more advanced stages, likely due to build-up of ethylene in the bags. An approach was developed to the processing of hyperspectral data for non-invasive monitoring of the lettuce leaves with a potential for implementation in greenhouses and packing lines.


RSC Advances ◽  
2016 ◽  
Vol 6 (66) ◽  
pp. 61771-61776 ◽  
Author(s):  
Yu Gao ◽  
Chenjie Xu ◽  
Lianhui Wang

Raman imaging allows the non-invasive and label-free monitoring of the preferred osteogenic differentiation of human mesenchymal stem cells on the polycaprolactone scaffolds.


Biomaterials ◽  
2013 ◽  
Vol 34 (34) ◽  
pp. 8607-8616 ◽  
Author(s):  
Tyler Chang ◽  
Maxwell S. Zimmerley ◽  
Kyle P. Quinn ◽  
Isabelle Lamarre-Jouenne ◽  
David L. Kaplan ◽  
...  

2014 ◽  
Vol 8 (8) ◽  
pp. 638-645 ◽  
Author(s):  
Qifei Li ◽  
Edison Suasnavas ◽  
Lifu Xiao ◽  
Sierra Heywood ◽  
Xiaojun Qi ◽  
...  

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