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1
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Mazarakis N, Vongsvivut J, Bambery KR, Ververis K, Tobin MJ, Royce SG, Samuel CS, Snibson KJ, Licciardi PV, Karagiannis TC. Investigation of molecular mechanisms of experimental compounds in murine models of chronic allergic airways disease using synchrotron Fourier-transform infrared microspectroscopy. Sci Rep 2020; 10:11713. [PMID: 32678217 PMCID: PMC7366655 DOI: 10.1038/s41598-020-68671-2] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/25/2019] [Accepted: 06/29/2020] [Indexed: 12/31/2022] Open
Abstract
The ovalbumin-induced (OVA) chronic allergic airways murine model is a well-established model for investigating pre-clinical therapies for chronic allergic airways diseases, such as asthma. Here, we examined the effects of several experimental compounds with potential anti-asthmatic effects including resveratrol (RV), relaxin (RLN), l-sulforaphane (LSF), valproic acid (VPA), and trichostatin A (TSA) using both a prevention and reversal model of chronic allergic airways disease. We undertook a novel analytical approach using focal plane array (FPA) and synchrotron Fourier-transform infrared (S-FTIR) microspectroscopic techniques to provide new insights into the mechanisms of action of these experimental compounds. Apart from the typical biological effects, S-FTIR microspectroscopy was able to detect changes in nucleic acids and protein acetylation. Further, we validated the reduction in collagen deposition induced by each experimental compound evaluated. Although this has previously been observed with conventional histological methods, the S-FTIR technique has the advantage of allowing identification of the type of collagen present. More generally, our findings highlight the potential utility of S-FTIR and FPA-FTIR imaging techniques in enabling a better mechanistic understanding of novel asthma therapeutics.
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Affiliation(s)
- Nadia Mazarakis
- Epigenomic Medicine Laboratory, Department of Diabetes, Central Clinical School, Monash University, Alfred Centre, 99 Commercial Road, Melbourne, VIC, 3004, Australia.,Faculty of Veterinary and Agricultural Sciences, University of Melbourne, Parkville, VIC, 3010, Australia.,Murdoch Children's Research Institute, Melbourne, VIC, 3004, Australia
| | | | | | - Katherine Ververis
- Epigenomic Medicine Laboratory, Department of Diabetes, Central Clinical School, Monash University, Alfred Centre, 99 Commercial Road, Melbourne, VIC, 3004, Australia
| | - Mark J Tobin
- ANSTO Australian Synchrotron, Clayton, VIC, 3168, Australia
| | - Simon G Royce
- Department of Pharmacology, Monash Biomedicine Discovery Institute, Monash University, Clayton, VIC, 3168, Australia
| | - Chrishan S Samuel
- Department of Pharmacology, Monash Biomedicine Discovery Institute, Monash University, Clayton, VIC, 3168, Australia
| | - Kenneth J Snibson
- Faculty of Veterinary and Agricultural Sciences, University of Melbourne, Parkville, VIC, 3010, Australia
| | - Paul V Licciardi
- Murdoch Children's Research Institute, Melbourne, VIC, 3004, Australia.,Department of Paediatrics, University of Melbourne, Parkville, VIC, 3010, Australia
| | - Tom C Karagiannis
- Epigenomic Medicine Laboratory, Department of Diabetes, Central Clinical School, Monash University, Alfred Centre, 99 Commercial Road, Melbourne, VIC, 3004, Australia. .,Department of Clinical Pathology, University of Melbourne, Parkville, VIC, 3010, Australia.
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Zafari J, Jouni FJ, Ahmadvand A, Abdolmaleki P, Soodi M, Zendehdel R. Investigation of gene expressions in differentiated cell derived bone marrow stem cells during bone morphogenetic protein-4 treatments with Fourier transform infrared spectroscopy. SPECTROCHIMICA ACTA. PART A, MOLECULAR AND BIOMOLECULAR SPECTROSCOPY 2017; 173:695-703. [PMID: 27780130 DOI: 10.1016/j.saa.2016.10.015] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 06/27/2015] [Revised: 09/17/2016] [Accepted: 10/16/2016] [Indexed: 06/06/2023]
Abstract
A model was set up to predict the differentiation patterns based on the data extracted from FTIR spectroscopy. For this reason, bone marrow stem cells (BMSCs) were differentiated to primordial germ cells (PGCs). Changes in cellular macromolecules in the time of 0, 24, 48, 72, and 96h of differentiation, as different steps of the differentiation procedure were investigated by using FTIR spectroscopy. Also, the expression of pluripotency (Oct-4, Nanog and c-Myc) and specific genes (Mvh, Stella and Fragilis) were investigated by real-time PCR. However, the expression of genes in five steps of differentiation was predicted by FTIR spectroscopy. FTIR spectra showed changes in the template of band intensities at different differentiation steps. There are increasing changes in the stepwise differentiation procedure for the ratio area of CH2, which is symmetric to CH2 asymmetric stretching. An ensemble of expert methods, including regression tree (RT), boosting algorithm (BA), and generalized regression neural network (GRNN), was the best method to predict the gene expression by FTIR spectroscopy. In conclusion, the model was able to distinguish the pattern of different steps from cell differentiation by using some useful features extracted from FTIR spectra.
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Affiliation(s)
- Jaber Zafari
- Department of Toxicology and Pharmacology, Faculty of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
| | - Fatemeh Javani Jouni
- Department of Microbiology, Islamic Azad University, Tehran North Branch, Tehran, Iran.
| | - Ali Ahmadvand
- Computer Science and Informatics Department, EMORY University School of Medicine, Atlanta, GA, USA.
| | - Parviz Abdolmaleki
- Department of Microbiology, Islamic Azad University, Tehran North Branch, Tehran, Iran.
| | - Malihe Soodi
- Department of Toxicology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
| | - Rezvan Zendehdel
- Environmental and Occupational Hazards control Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Department of Occupational Hygiene, School of Public Health, Shahid Beheshti University, Tehran, Iran..
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Werkmeister RM, Sapeta S, Schmidl D, Garhöfer G, Schmidinger G, Aranha dos Santos V, Aschinger GC, Baumgartner I, Pircher N, Schwarzhans F, Pantalon A, Dua H, Schmetterer L. Ultrahigh-resolution OCT imaging of the human cornea. BIOMEDICAL OPTICS EXPRESS 2017; 8:1221-1239. [PMID: 28271013 PMCID: PMC5330598 DOI: 10.1364/boe.8.001221] [Citation(s) in RCA: 67] [Impact Index Per Article: 8.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 11/07/2016] [Revised: 12/19/2016] [Accepted: 12/26/2016] [Indexed: 05/03/2023]
Abstract
We present imaging of corneal pathologies using optical coherence tomography (OCT) with high resolution. To this end, an ultrahigh-resolution spectral domain OCT (UHR-OCT) system based on a broad bandwidth Ti:sapphire laser is employed. With a central wavelength of 800 nm, the imaging device allows to acquire OCT data at the central, paracentral and peripheral cornea as well as the limbal region with 1.2 µm x 20 µm (axial x lateral) resolution at a rate of 140 000 A-scans/s. Structures of the anterior segment of the eye, not accessible with commercial OCT systems, are visualized. These include corneal nerves, limbal palisades of Vogt as well as several corneal pathologies. Cases such as keratoconus and Fuchs's endothelial dystrophy as well as infectious changes caused by diseases like Acanthamoeba keratitis and scarring after herpetic keratitis are presented. We also demonstrate the applicability of our system to visualize epithelial erosion and intracorneal foreign body after corneal trauma as well as chemical burns. Finally, results after Descemet's membrane endothelial keratoplasty (DMEK) are imaged. These clinical cases show the potential of UHR-OCT to help in clinical decision-making and follow-up. Our results and experience indicate that UHR-OCT of the cornea is a promising technique for the use in clinical practice, but can also help to gain novel insight in the physiology and pathophysiology of the human cornea.
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Affiliation(s)
- René M. Werkmeister
- Center for Medical Physics and Biomedical Engineering, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
- Christian Doppler Laboratory for Ocular and Dermal Effects of Thiomers, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
| | - Sabina Sapeta
- Center for Medical Physics and Biomedical Engineering, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
- Singapore Eye Research Institute The Academia, 20 College Road, Discovery Tower Level 6, 169856, Singapore
| | - Doreen Schmidl
- Department of Clinical Pharmacology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
| | - Gerhard Garhöfer
- Christian Doppler Laboratory for Ocular and Dermal Effects of Thiomers, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
- Department of Clinical Pharmacology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
| | - Gerald Schmidinger
- Department of Ophthalmology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
| | - Valentin Aranha dos Santos
- Center for Medical Physics and Biomedical Engineering, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
| | - Gerold C. Aschinger
- Center for Medical Physics and Biomedical Engineering, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
| | - Isabella Baumgartner
- Department of Ophthalmology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
| | - Niklas Pircher
- Department of Ophthalmology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
| | - Florian Schwarzhans
- Department of Ophthalmology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
- Center for Medical Statistics, Informatics and Intelligent Systems, Medical University of Vienna, Spitalgasse 23, A-1090 Vienna, Austria
| | - Anca Pantalon
- Department of Ophthalmology, Gr. T. Popa University of Medicine and Pharmacy, Iasi, Sf. Spiridon University Hospital, 16 Universitatii Str, Iasi, 700115, Romania
| | - Harminder Dua
- Academic Section of Ophthalmology, Division of Clinical Neuroscience, Nottingham University Hospitals NHS Trust QMC campus, Derby Road, Nottingham, NG7 2UH, UK
| | - Leopold Schmetterer
- Center for Medical Physics and Biomedical Engineering, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
- Christian Doppler Laboratory for Ocular and Dermal Effects of Thiomers, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
- Singapore Eye Research Institute The Academia, 20 College Road, Discovery Tower Level 6, 169856, Singapore
- Department of Clinical Pharmacology, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna, Austria
- Lee Kong Chian School of Medicine, Nanyang Technological University Novena Campus, 11 Mandalay Road, 308232, Singapore
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Shen HH, Liu GS, Chow SH, Wang JH, He Z, Nguyen C, Lin TW, Bui BV. Intraocular Pressure Induced Retinal Changes Identified Using Synchrotron Infrared Microscopy. PLoS One 2016; 11:e0164035. [PMID: 27711151 PMCID: PMC5053542 DOI: 10.1371/journal.pone.0164035] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/04/2016] [Accepted: 09/19/2016] [Indexed: 11/18/2022] Open
Abstract
Infrared (IR) spectroscopy has been used to quantify chemical and structural characteristics of a wide range of materials including biological tissues. In this study, we examined spatial changes in the chemical characteristics of rat retina in response to intraocular pressure (IOP) elevation using synchrotron infrared microscopy (SIRM), a non-destructive imaging approach. IOP elevation was induced by placing a suture around the eye of anaesthetised rats. Retinal sections were collected onto transparent CaF2 slides 10 days following IOP elevation. Using combined SIRM spectra and chemical mapping approaches it was possible to quantify IOP induced changes in protein conformation and chemical distribution in various layers of the rat retina. We showed that 10 days following IOP elevation there was an increase in lipid and protein levels in the inner nuclear layer (INL) and ganglion cell layer (GCL). IOP elevation also resulted in an increase in nucleic acids in the INL. Analysis of SIRM spectra revealed a shift in amide peaks to lower vibrational frequencies with a more prominent second shoulder, which is consistent with the presence of cell death in specific layers of the retina. These changes were more substantial in the INL and GCL layers compared with those occurring in the outer nuclear layer. These outcomes demonstrate the utility of SIRM to quantify the effect of IOP elevation on specific layers of the retina. Thus SIRM may be a useful tool for the study of localised tissue changes in glaucoma and other eye diseases.
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Affiliation(s)
- Hsin-Hui Shen
- Department of Materials Science and Engineering, Faculty of Engineering, Monash University, Clayton, Victoria, Australia
- Infection and Immunity Program, Biomedicine Discovery Institute and Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia
- * E-mail: (HHS); (BVB)
| | - Guei-Sheung Liu
- Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia
- Ophthalmology, Department of Surgery, University of Melbourne, East Melbourne, Victoria, Australia
| | - Seong Hoong Chow
- Infection and Immunity Program, Biomedicine Discovery Institute and Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia
| | - Jiang-Hui Wang
- Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia
- Ophthalmology, Department of Surgery, University of Melbourne, East Melbourne, Victoria, Australia
| | - Zheng He
- Department of Optometry & Vision Sciences, University of Melbourne, Parkville, Victoria, Australia
| | - Christine Nguyen
- Department of Optometry & Vision Sciences, University of Melbourne, Parkville, Victoria, Australia
| | - Tsung-Wu Lin
- Department of Chemistry, Tunghai University, Taichung City, Taiwan
| | - Bang V. Bui
- Department of Optometry & Vision Sciences, University of Melbourne, Parkville, Victoria, Australia
- * E-mail: (HHS); (BVB)
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5
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Kalmodia S, Parameswaran S, Yang W, Barrow CJ, Krishnakumar S. Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy: An analytical technique to understand therapeutic responses at the molecular level. Sci Rep 2015; 5:16649. [PMID: 26568521 PMCID: PMC4645174 DOI: 10.1038/srep16649] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/21/2015] [Accepted: 10/14/2015] [Indexed: 02/07/2023] Open
Abstract
Rapid monitoring of the response to treatment in cancer patients is essential to predict the outcome of the therapeutic regimen early in the course of the treatment. The conventional methods are laborious, time-consuming, subjective and lack the ability to study different biomolecules and their interactions, simultaneously. Since; mechanisms of cancer and its response to therapy is dependent on molecular interactions and not on single biomolecules, an assay capable of studying molecular interactions as a whole, is preferred. Fourier Transform Infrared (FTIR) spectroscopy has become a popular technique in the field of cancer therapy with an ability to elucidate molecular interactions. The aim of this study, was to explore the utility of the FTIR technique along with multivariate analysis to understand whether the method has the resolution to identify the differences in the mechanism of therapeutic response. Towards achieving the aim, we utilized the mouse xenograft model of retinoblastoma and nanoparticle mediated targeted therapy. The results indicate that the mechanism underlying the response differed between the treated and untreated group which can be elucidated by unique spectral signatures generated by each group. The study establishes the efficiency of non-invasive, label-free and rapid FTIR method in assessing the interactions of nanoparticles with cellular macromolecules towards monitoring the response to cancer therapeutics.
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Affiliation(s)
- Sushma Kalmodia
- Department of Nano biotechnology, Vision Research Foundation, Sankara Nethralaya, 18, College Road, Nungambakkam, Chennai - 600 006, India.,Centre for Chemistry and Biotechnology, Deakin University, Geelong campus, VIC 3216, Australia
| | - Sowmya Parameswaran
- Radheshyam Kanoi Stem Cell laboratory, Vision Research Foundation, Sankara Nethralaya, 18, College Road, Nungambakkam, Chennai - 600 006, India
| | - Wenrong Yang
- Centre for Chemistry and Biotechnology, Deakin University, Geelong campus, VIC 3216, Australia
| | - Colin J Barrow
- Centre for Chemistry and Biotechnology, Deakin University, Geelong campus, VIC 3216, Australia
| | - Subramanian Krishnakumar
- Department of Nano biotechnology, Vision Research Foundation, Sankara Nethralaya, 18, College Road, Nungambakkam, Chennai - 600 006, India
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6
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Yoon JJ, Ismail S, Sherwin T. Limbal stem cells: Central concepts of corneal epithelial homeostasis. World J Stem Cells 2014; 6:391-403. [PMID: 25258661 PMCID: PMC4172668 DOI: 10.4252/wjsc.v6.i4.391] [Citation(s) in RCA: 81] [Impact Index Per Article: 7.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/25/2014] [Revised: 08/20/2014] [Accepted: 09/01/2014] [Indexed: 02/06/2023] Open
Abstract
A strong cohort of evidence exists that supports the localisation of corneal stem cells at the limbus. The distinguishing characteristics of limbal cells as stem cells include slow cycling properties, high proliferative potential when required, clonogenicity, absence of differentiation marker expression coupled with positive expression of progenitor markers, multipotency, centripetal migration, requirement for a distinct niche environment and the ability of transplanted limbal cells to regenerate the entire corneal epithelium. The existence of limbal stem cells supports the prevailing theory of corneal homeostasis, known as the XYZ hypothesis where X represents proliferation and stratification of limbal basal cells, Y centripetal migration of basal cells and Z desquamation of superficial cells. To maintain the mass of cornea, the sum of X and Y must equal Z and very elegant cell tracking experiments provide strong evidence in support of this theory. However, several recent studies have suggested the existence of oligopotent stem cells capable of corneal maintenance outside of the limbus. This review presents a summary of data which led to the current concepts of corneal epithelial homeostasis and discusses areas of controversy surrounding the existence of a secondary stem cell reservoir on the corneal surface
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7
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Patel II, Shearer DA, Fogarty SW, Fullwood NJ, Quaroni L, Martin FL, Weisz J. Infrared microspectroscopy identifies biomolecular changes associated with chronic oxidative stress in mammary epithelium and stroma of breast tissues from healthy young women: implications for latent stages of breast carcinogenesis. Cancer Biol Ther 2013; 15:225-35. [PMID: 24107651 DOI: 10.4161/cbt.26748] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/21/2022] Open
Abstract
Studies of the decades-long latent stages of breast carcinogenesis have been limited to when hyperplastic lesions are already present. Investigations of earlier stages of breast cancer (BC) latency have been stymied by the lack of fiducial biomarkers needed to identify where in histologically normal tissues progression toward a BC might be taking place. Recent evidence suggests that a marker of chronic oxidative stress (OxS), protein adducts of 4-hydroxy-2-nonenal (4HNE), can meet this need. Specifically: (1) 4HNE immunopositive (4HNE+) mammary epithelial (ME) cells were found to be prevalent in normal (reduction mammoplasty) tissues of most women (including many teenagers) studied, representative of those living in the United States' high risk-posing environment and: (2) marked (> 1.5-fold) differences were identified between tissues of healthy young women with many vs. few 4HNE+ ME cells in the relative levels of transcripts for 42 of the 84 OxS-associated genes represented in SABioscience Oxidative-Stress/Oxidative-Defense PCR array. Herein we used synchrotron radiation-based Fourier-transform infrared (SR-FTIR) microspectroscopy to identify molecular changes associated with 4HNE adducts in basal and luminal ME cells in terminal ductal units (TDLU), which are the cells of origin of BC, and associated intralobular and interlobular stroma, known contributors to carcinogenesis. Multivariate analysis-derived wavenumbers differentiated 4HNE+ and 4HNE- cells in each of the anatomical compartments. Specifically, principal component and linear discriminant analyses of mid-infrared spectra obtained from these cells revealed unambiguous, statistically highly significant differences in the "biochemical fingerprint" of 4HNE+ vs. 4HNE- luminal and basal ME cells, as well as between associated intralobular and interlobular stroma. These findings demonstrate further SR-FTIR microspectroscopy's ability to identify molecular changes associated with altered physiological and/or pathophysiological states, in this case with a state of chronic OxS that provides a pro-carcinogenic microenvironment.
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Affiliation(s)
- Imran I Patel
- Center for Biophotonics; Lancaster Environment Centre; Lancaster University; Lancaster, UK
| | - Debra A Shearer
- Department of Obstetrics and Gynecology; College of Medicine; Pennsylvania State University; Hershey, PA USA
| | - Simon W Fogarty
- Division of Biomedical and Life Sciences; Faculty of Health and Medicine; Lancaster University; Lancaster, UK
| | - Nigel J Fullwood
- Division of Biomedical and Life Sciences; Faculty of Health and Medicine; Lancaster University; Lancaster, UK
| | | | - Francis L Martin
- Center for Biophotonics; Lancaster Environment Centre; Lancaster University; Lancaster, UK
| | - Judith Weisz
- Department of Obstetrics and Gynecology; College of Medicine; Pennsylvania State University; Hershey, PA USA; Department of Pathology; College of Medicine; Pennsylvania State University; Hershey, PA USA
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Cao J, Ng ES, McNaughton D, Stanley EG, Elefanty AG, Tobin MJ, Heraud P. The characterisation of pluripotent and multipotent stem cells using Fourier transform infrared microspectroscopy. Int J Mol Sci 2013; 14:17453-76. [PMID: 24065090 PMCID: PMC3794735 DOI: 10.3390/ijms140917453] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/29/2013] [Revised: 07/22/2013] [Accepted: 07/22/2013] [Indexed: 01/08/2023] Open
Abstract
Fourier transform infrared (FTIR) microspectroscopy shows potential as a benign, objective and rapid tool to screen pluripotent and multipotent stem cells for clinical use. It offers a new experimental approach that provides a holistic measurement of macromolecular composition such that a signature representing the internal cellular phenotype is obtained. The use of this technique therefore contributes information that is complementary to that acquired by conventional genetic and immunohistochemical methods.
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Affiliation(s)
- Julie Cao
- Department of Anatomy and Developmental Biology, Faculty of Medicine, Nursing and Health Sciences, Monash University, Wellington Road, Clayton, Victoria 3800, Australia; E-Mails: (J.C.); (E.S.N.); (E.G.S.); (A.G.E.)
- Centre for Biospectroscopy and the School of Chemistry, Monash University, Clayton, Victoria 3800, Australia; E-Mail:
| | - Elizabeth S. Ng
- Department of Anatomy and Developmental Biology, Faculty of Medicine, Nursing and Health Sciences, Monash University, Wellington Road, Clayton, Victoria 3800, Australia; E-Mails: (J.C.); (E.S.N.); (E.G.S.); (A.G.E.)
- Murdoch Childrens Research Institute, the Royal Children’s Hospital, Parkville, Victoria 3052, Australia
| | - Donald McNaughton
- Centre for Biospectroscopy and the School of Chemistry, Monash University, Clayton, Victoria 3800, Australia; E-Mail:
| | - Edouard G. Stanley
- Department of Anatomy and Developmental Biology, Faculty of Medicine, Nursing and Health Sciences, Monash University, Wellington Road, Clayton, Victoria 3800, Australia; E-Mails: (J.C.); (E.S.N.); (E.G.S.); (A.G.E.)
- Murdoch Childrens Research Institute, the Royal Children’s Hospital, Parkville, Victoria 3052, Australia
| | - Andrew G. Elefanty
- Department of Anatomy and Developmental Biology, Faculty of Medicine, Nursing and Health Sciences, Monash University, Wellington Road, Clayton, Victoria 3800, Australia; E-Mails: (J.C.); (E.S.N.); (E.G.S.); (A.G.E.)
- Murdoch Childrens Research Institute, the Royal Children’s Hospital, Parkville, Victoria 3052, Australia
| | - Mark J. Tobin
- Australian Synchrotron, 800 Blackburn Road, Clayton, Victoria 3168, Australia; E-Mail:
| | - Philip Heraud
- Department of Anatomy and Developmental Biology, Faculty of Medicine, Nursing and Health Sciences, Monash University, Wellington Road, Clayton, Victoria 3800, Australia; E-Mails: (J.C.); (E.S.N.); (E.G.S.); (A.G.E.)
- Centre for Biospectroscopy and the School of Chemistry, Monash University, Clayton, Victoria 3800, Australia; E-Mail:
- Author to whom correspondence should be addressed; E-Mail: ; Tel.: +61-3-9905-0765; Fax: +61-3-9905-5613
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Clemens G, Flower KR, Henderson AP, Whiting A, Przyborski SA, Jimenez-Hernandez M, Ball F, Bassan P, Cinque G, Gardner P. The action of all-trans-retinoic acid (ATRA) and synthetic retinoid analogues (EC19 and EC23) on human pluripotent stem cells differentiation investigated using single cell infrared microspectroscopy. MOLECULAR BIOSYSTEMS 2013; 9:677-92. [PMID: 23364809 DOI: 10.1039/c3mb25505k] [Citation(s) in RCA: 26] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/21/2022]
Abstract
All trans-retinoic acid (ATRA) is widely used to direct the differentiation of cultured stem cells. When exposed to the pluripotent human embryonal carcinoma (EC) stem cell line, TERA2.cl.SP12, ATRA induces ectoderm differentiation and the formation of neuronal cell types. We have previously generated synthetic analogues of retinoic acid (EC23 and EC19) which also induce the differentiation of EC cells. Even though EC23 and EC19 have similar chemical structures, they have differing biochemical effects in terms of EC cell differentiation. EC23 induces neuronal differentiation in a manner similar to ATRA, whereas EC19 directs the cells to form epithelial-like derivatives. Previous MALDI-TOF MS analysis examined the response of TERA2.cl.SP12 cells after exposure to ATRA, EC23 and EC19 and further demonstrated the similarly in the effect of ATRA and EC23 activity whilst responses to EC19 were very different. In this study, we show that Fourier Transform Infrared Micro-Spectroscopy (FT-IRMS) coupled with appropriate scatter correction and multivariate analysis can be used as an effective tool to further investigate the differentiation of human pluripotent stem cells and monitor the alternative affects different retinoid compounds have on the induction of differentiation. FT-IRMS detected differences between cell populations as early as 3 days of compound treatment. Populations of cells treated with different retinoid compounds could easily be distinguished from one another during the early stages of cell differentiation. These data demonstrate that FT-IRMS technology can be used as a sensitive screening technique to monitor the status of the stem cell phenotype and progression of differentiation along alternative pathways in response to different compounds.
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Affiliation(s)
- Graeme Clemens
- Manchester Institute of Biotechnology, Manchester University, 131 Princess Street, Manchester, M1 7DN, UK
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10
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Oral cancer diagnostics based on infrared spectral markers and wax physisorption kinetics. Anal Bioanal Chem 2013; 405:1995-2007. [PMID: 23318761 DOI: 10.1007/s00216-012-6625-z] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/05/2012] [Revised: 11/13/2012] [Accepted: 12/03/2012] [Indexed: 10/27/2022]
Abstract
Infrared microspectroscopy is an emerging approach for disease analysis owing to its capability for in situ chemical characterization of pathological processes. Synchrotron-based infrared microspectroscopy (SR-IMS) provides ultra-high spatial resolution for profiling biochemical events associated with disease progression. Spectral alterations were observed in cultured oral cells derived from healthy, precancerous, primary, and metastatic cancers. An innovative wax-physisorption-based kinetic FTIR imaging method for the detection of oral precancer and cancer was demonstrated successfully. The approach is based on determining the residual amount of paraffin wax (C(25)H(52)) or beeswax (C(46)H(92)O(2)) on a sample surface after xylene washing. This amount is used as a signpost of the degree of physisorption that altered during malignant transformation. The results of linear discriminant analysis (LDA) of oral cell lines indicated that the methylene (CH(2)) and methyl group (CH(3)) stretching vibrations in the range of 3,000-2,800 cm(-1) have the highest accuracy rate (89.6 %) to discriminate the healthy keratinocytes (NHOK) from cancer cells. The results of wax-physisorption-based FTIR imaging showed a stronger physisorption with beeswax in oral precancerous and cancer cells as compared with that of NHOK, which showed a strong capability with paraffin wax. The infrared kinetic study of oral cavity tissue showed a consistency in the wax physisorption of the cell lines. On the basis of our findings, these results show the potential use of wax-physisorption-based kinetic FTIR imaging for the early screening of oral cancer lesions and the chemical changes during oral carcinogenesis.
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11
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Cao J, Ng ES, McNaughton D, Stanley EG, Elefanty AG, Tobin MJ, Heraud P. Fourier transform infrared microspectroscopy reveals that tissue culture conditions affect the macromolecular phenotype of human embryonic stem cells. Analyst 2013; 138:4147-60. [DOI: 10.1039/c3an00321c] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/21/2022]
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12
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Fogarty SW, Patel II, Trevisan J, Nakamura T, Hirschmugl CJ, Fullwood NJ, Martin FL. Sub-cellular spectrochemical imaging of isolated human corneal cells employing synchrotron radiation-based Fourier-transform infrared microspectroscopy. Analyst 2013; 138:240-8. [DOI: 10.1039/c2an36197c] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022]
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13
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Sandt C, Féraud O, Oudrhiri N, Bonnet ML, Meunier MC, Valogne Y, Bertrand A, Raphaël M, Griscelli F, Turhan AG, Dumas P, Bennaceur-Griscelli A. Identification of spectral modifications occurring during reprogramming of somatic cells. PLoS One 2012; 7:e30743. [PMID: 22514597 PMCID: PMC3326006 DOI: 10.1371/journal.pone.0030743] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/27/2011] [Accepted: 12/25/2011] [Indexed: 11/18/2022] Open
Abstract
Recent technological advances in cell reprogramming by generation of induced pluripotent stem cells (iPSC) offer major perspectives in disease modelling and future hopes for providing novel stem cells sources in regenerative medicine. However, research on iPSC still requires refining the criteria of the pluripotency stage of these cells and exploration of their equivalent functionality to human embryonic stem cells (ESC). We report here on the use of infrared microspectroscopy to follow the spectral modification of somatic cells during the reprogramming process. We show that induced pluripotent stem cells (iPSC) adopt a chemical composition leading to a spectral signature indistinguishable from that of embryonic stem cells (ESC) and entirely different from that of the original somatic cells. Similarly, this technique allows a distinction to be made between partially and fully reprogrammed cells. We conclude that infrared microspectroscopy signature is a novel methodology to evaluate induced pluripotency and can be added to the tests currently used for this purpose.
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Affiliation(s)
| | - Olivier Féraud
- Inserm UMR 935, “ESTeam Paris Sud,” Stem Cell Core Facility Institut André Lwoff, University Paris Sud 11 Paul Brousse, Villejuif, France
| | - Noufissa Oudrhiri
- Inserm UMR 935, “ESTeam Paris Sud,” Stem Cell Core Facility Institut André Lwoff, University Paris Sud 11 Paul Brousse, Villejuif, France
- AP-HP Laboratory of Hematology, CHU, Bicêtre, France
| | - Marie Laure Bonnet
- Inserm UMR 935, “ESTeam Paris Sud,” Stem Cell Core Facility Institut André Lwoff, University Paris Sud 11 Paul Brousse, Villejuif, France
- Inserm UMR 935, University of Poitiers, Division of Laboratory Hematology and Oncology, CHU Poitiers, Poitiers, France
| | - Marie Claude Meunier
- Inserm UMR 935, “ESTeam Paris Sud,” Stem Cell Core Facility Institut André Lwoff, University Paris Sud 11 Paul Brousse, Villejuif, France
- Inserm UMR 935, University of Poitiers, Division of Laboratory Hematology and Oncology, CHU Poitiers, Poitiers, France
| | - Yannick Valogne
- Inserm UMR 935, “ESTeam Paris Sud,” Stem Cell Core Facility Institut André Lwoff, University Paris Sud 11 Paul Brousse, Villejuif, France
| | - Angelina Bertrand
- Inserm UMR 935, “ESTeam Paris Sud,” Stem Cell Core Facility Institut André Lwoff, University Paris Sud 11 Paul Brousse, Villejuif, France
- Inserm UMR 935, University of Poitiers, Division of Laboratory Hematology and Oncology, CHU Poitiers, Poitiers, France
| | | | - Frank Griscelli
- Inserm UMR 935, “ESTeam Paris Sud,” Stem Cell Core Facility Institut André Lwoff, University Paris Sud 11 Paul Brousse, Villejuif, France
- University Paris Descartes, Paris, France
| | - Ali G. Turhan
- Inserm UMR 935, “ESTeam Paris Sud,” Stem Cell Core Facility Institut André Lwoff, University Paris Sud 11 Paul Brousse, Villejuif, France
- Inserm UMR 935, University of Poitiers, Division of Laboratory Hematology and Oncology, CHU Poitiers, Poitiers, France
- * E-mail:
| | - Paul Dumas
- SOLEIL Synchrotron, Saint Aubin, Gif sur Yvette, France
| | - Annelise Bennaceur-Griscelli
- Inserm UMR 935, “ESTeam Paris Sud,” Stem Cell Core Facility Institut André Lwoff, University Paris Sud 11 Paul Brousse, Villejuif, France
- AP-HP Laboratory of Hematology, CHU, Bicêtre, France
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14
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Effects on antigen-presenting cells of short-term interaction with the human host defence peptide β-defensin 2. Biochem J 2011; 436:537-46. [DOI: 10.1042/bj20101977] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/17/2022]
Abstract
β-Defensins are antimicrobial peptides that exert their host-defence functions at the interface between the host and microbial biota. They display a direct, salt- and medium-sensitive cidal activity, in vitro, against a broad spectrum of bacteria and fungi, and there is increasing evidence that they also play a role in alerting and enhancing cellular components of innate and adaptive immunity. Their interaction with biological membranes plays a central role in both of these types of activities. In the present study, we have investigated the interaction of fluorescently labelled hBD2 (human β-defensin 2) with monocytes, macrophages and iDCs (immature dendritic cells), observing a differential capacity to be rapidly internalized into these cells. Complementary microscopy techniques [TEM (transmission electron microscopy), optical microscopy and IR microspectroscopy] were used to explore the functional and biological implications of these interactions on iDCs. Short-term exposure to the peptide resulted in significant alterations in membrane composition and re-organization of the endomembrane system, with the induction of degranulation. These events may be associated with the antigen-presenting activities or the chemotaxis of iDCs, which appears to occur via both CCR6 (CC chemokine receptor 6)-dependent and -independent mechanisms.
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15
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Ami D, Mereghetti P, Natalello A, Doglia SM, Zanoni M, Redi CA, Monti M. FTIR spectral signatures of mouse antral oocytes: molecular markers of oocyte maturation and developmental competence. BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH 2011; 1813:1220-9. [PMID: 21435359 DOI: 10.1016/j.bbamcr.2011.03.009] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/12/2010] [Revised: 02/15/2011] [Accepted: 03/15/2011] [Indexed: 12/11/2022]
Abstract
Mammalian antral oocytes with a Hoescht-positive DNA ring around the nucleolus (SN) are able to resume meiosis and to fully support the embryonic development, while oocytes with a non-surrounded nucleolus (NSN) cannot. Here, we applied FTIR microspectroscopy to characterize single SN and NSN mouse oocytes in order to try to elucidate some aspects of the mechanisms behind the different chromatin organization that impairs the full development of NSN oocyte-derived embryos. To this aim, oocytes were measured at three different stages of their maturation: just after isolation and classification as SN and NSN oocytes (time 0); after 10h of in vitro maturation, i.e. at the completion of the metaphase I (time 1); and after 20h of in vitro maturation, i.e. at the completion of the metaphase II (time 2). Significant spectral differences in the lipid (3050-2800cm(-1)) and protein (1700-1600cm(-1)) absorption regions were found between the two types of oocytes and among the different stages of maturation within the same oocyte type. Moreover, dramatic changes in nucleic acid content, concerning mainly the extent of transcription and polyadenylation, were detected in particular between 1000 and 800cm(-1). The use of the multivariate principal component-linear discriminant analysis (PCA-LDA) enabled us to identify the maturation stage in which the separation between the two types of oocytes took place, finding as the most discriminating wavenumbers those associated to transcriptional activity and polyadenylation, in agreement with the visual analysis of the spectral data.
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Affiliation(s)
- Diletta Ami
- Fondazione IRCCS Policlinico San Matteo, V.le C. Golgi 19, Pavia, Italy
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16
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Ma A, Zhao B, Bentley AJ, Brahma A, MacNeil S, Martin FL, Rimmer S, Fullwood NJ. Corneal epithelialisation on surface-modified hydrogel implants: artificial cornea. JOURNAL OF MATERIALS SCIENCE. MATERIALS IN MEDICINE 2011; 22:663-670. [PMID: 21287242 DOI: 10.1007/s10856-011-4244-4] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 07/19/2010] [Accepted: 01/17/2011] [Indexed: 05/30/2023]
Abstract
The objective was to investigate corneal re-epithelialisation of surface-modified polymethacrylate hydrogel implants in order to evaluate them as potential materials for an artificial cornea. Polymethacrylate hydrogels were modified with amines and then coated with different extracellular matrix proteins (collagen I, IV, laminin and fibronectin). The modified hydrogels were surgically implanted into bovine corneas maintained in a 3-D culture system for 5 days. The epithelial growth across the implant surface was evaluated using fluorescent, light and electron microscopy. Full epithelialisation was achieved on 1,4-diaminobutane-modified hydrogels after coating with collagen IV. Hydrogels modified with 1,4-diaminobutane but without further coating only showed partial re-epithelialisation. Hydrogels modified with other amines (1,2-diaminoethane or 1,3-diaminopropane) showed only partial re-epithelialisation; further coating with extracellular matrix proteins improved epithelialisation of these surfaces but did not result in complete re-epithelialisation. Evaluation of the corneas implanted with the 1,4-diaminobutane-modified hydrogels coated with collagen IV showed that the artificial corneas remain clear, integrate well and become covered by a healthy stratified epithelium. In conclusion the 1,4-diaminobutane surface-modified hydrogel coated with collagen IV supported the growth of a stable stratified epithelium. With further refinement this hydrogel has the potential to be used clinically for an artificial cornea.
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Affiliation(s)
- Aihua Ma
- School of Health and Medicine, Biomedical and Life Sciences, Lancaster University, Lancaster, UK
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17
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Lu R, Bian F, Zhang X, Qi H, Chuang EY, Pflugfelder SC, Li DQ. The β-catenin/Tcf4/survivin signaling maintains a less differentiated phenotype and high proliferative capacity of human corneal epithelial progenitor cells. Int J Biochem Cell Biol 2011; 43:751-9. [PMID: 21292023 DOI: 10.1016/j.biocel.2011.01.018] [Citation(s) in RCA: 32] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/01/2010] [Revised: 01/18/2011] [Accepted: 01/24/2011] [Indexed: 01/07/2023]
Abstract
It is clear that the microenvironment or niche plays an important role in determining the fate of stem cells: being stem cells or differentiated. However, the intrinsic pathways controlling the fate of adult stem cells in different niches are largely unknown. This study was to explore the role of β-catenin/Tcf4/survivin signaling in determining the fate of human corneal epithelial stem cells in different media. We observed that the low calcium serum-free media, especially CnT-20, promoted proliferative capacity, colony forming efficiency and stem cell-like phenotype of human corneal epithelial cells (HCECs) when compared with the cells cultured in a high calcium serum-containing medium SHEM. Three key factors in Wnt signaling, β-catenin, Tcf4 and survivin, were found to be expressed higher by HCECs grown in CnT-20 than those cultured in SHEM, as evaluated by real-time PCR, Western blotting and immunostaining. Transfection of siRNA-Tcf4 at 10-50nM knocked down Tcf4, and also significantly suppressed its down stream molecule survivin at both mRNA and protein levels in HCECs. Furthermore, Tcf4 silencing significantly suppressed the proliferative capacity of HCECs, measured by WST-1 assay, compared with the control groups, untreated or transfected with non-coding sequence siRNA-fluorescein. These findings demonstrate that low calcium serum free media promote ex vivo expansion of corneal epithelial progenitor cells that retain a less differentiated phenotype and high proliferative capacity via β-catenin/Tcf4/survivin signaling, a novel intrinsic pathway. This study may have high impact and clinic implication on the expansion of corneal epithelial stem cells in regenerative medicine, especially for ocular surface reconstruction.
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Affiliation(s)
- Rong Lu
- Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX, USA
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18
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Pijanka JK, Kumar D, Dale T, Yousef I, Parkes G, Untereiner V, Yang Y, Dumas P, Collins D, Manfait M, Sockalingum GD, Forsyth NR, Sulé-Suso J. Vibrational spectroscopy differentiates between multipotent and pluripotent stem cells. Analyst 2010; 135:3126-32. [PMID: 20953512 DOI: 10.1039/c0an00525h] [Citation(s) in RCA: 49] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022]
Abstract
Over the last few years, there has been an increased interest in the study of stem cells in biomedicine for therapeutic use and as a source for healing diseased or injured organs/tissues. More recently, vibrational spectroscopy has been applied to study stem cell differentiation. In this study, we have used both synchrotron based FTIR and Raman microspectroscopies to assess possible differences between human pluripotent (embryonic) and multipotent (adult mesenchymal) stem cells, and how O(2) concentration in cell culture could affect the spectral signatures of these cells. Our work shows that infrared spectroscopy of embryonic (pluripotent) and adult mesenchymal (multipotent) stem cells have different spectral signatures based on the amount of lipids in their cytoplasm (confirmed with cytological staining). Furthermore, O(2) concentration in cell culture causes changes in both the FTIR and Raman spectra of embryonic stem cells. These results show that embryonic stem cells might be more sensitive to O(2) concentration when compared to mesenchymal stem cells. While vibrational spectroscopy could therefore be of potential use in identifying different populations of stem cells further work is required to better understand these differences.
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Affiliation(s)
- Jacek Klaudiusz Pijanka
- Institute for Science and Technology in Medicine, Guy Hilton Research Centre, Keele University, Stoke on Trent, UK
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19
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Miller LM, Dumas P. From structure to cellular mechanism with infrared microspectroscopy. Curr Opin Struct Biol 2010; 20:649-56. [PMID: 20739176 DOI: 10.1016/j.sbi.2010.07.007] [Citation(s) in RCA: 83] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/31/2010] [Revised: 07/23/2010] [Accepted: 07/23/2010] [Indexed: 10/19/2022]
Abstract
Current efforts in structural biology aim to integrate structural information within the context of cellular organization and function. X-rays and infrared radiation stand at opposite ends of the electromagnetic spectrum and act as complementary probes for achieving this goal. Intense and bright beams are produced by synchrotron radiation, and are efficiently used in the wavelength domain extending from hard X-rays to the far-infrared (or THz) regime. While X-ray crystallography provides exquisite details on atomic structure, Fourier transform infrared microspectroscopy (FTIRM) is emerging as a spectroscopic probe and imaging tool for correlating molecular structure to biochemical dynamics and function. In this manuscript, the role of synchrotron FTIRM in bridging the gap towards 'functional biology' is discussed based upon recent achievements, with a critical assessment of the contributions to biological and biomedical research.
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Affiliation(s)
- Lisa M Miller
- National Synchrotron Light Source, Brookhaven National Laboratory, Upton, NY, USA.
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20
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Zelig U, Dror Z, Iskovich S, Zwielly A, Ben-Harush M, Nathan I, Mordechai S, Kapelushnik J. Biochemical analysis and quantification of hematopoietic stem cells by infrared spectroscopy. JOURNAL OF BIOMEDICAL OPTICS 2010; 15:037008. [PMID: 20615037 DOI: 10.1117/1.3442728] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/29/2023]
Abstract
Identification of hematopoietic stem cells (HSCs) in different stages of maturation is one of the major issues in stem cell research and bone marrow (BM) transplantation. Each stage of maturation of HSCs is characterized by a series of distinct glycoproteins present on the cell plasma membrane surface, named a cluster of differentiation (CD). Currently, complicated and expensive procedures based on CD expression are needed for identification and isolation of HSCs. This method is under dispute, since the correct markers' composition is not strictly clear, thus there is need for a better method for stem cell characterization. In the present study, Fourier transform infrared (FTIR) spectroscopy is employed as a novel optical method for identification and characterization of HSCs based on their entire biochemical features. FTIR spectral analysis of isolated mice HSCs reveals several spectral markers related to lipids, nucleic acids, and carbohydrates, which distinguish HSCs from BM cells. The unique "open" conformation of HSC DNA as identified by FTIR is exploited for HSCs quantification in the BM. The proposed method of FTIR spectroscopy for HSC identification and quantification can contribute to stem cell research and BM transplantation.
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Affiliation(s)
- Udi Zelig
- Ben-Gurion University of the Negev, Department of Biomedical Engineering, Beer-Sheva 84105 Israel
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21
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22
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Optical spectroscopy for noninvasive monitoring of stem cell differentiation. J Biomed Biotechnol 2010; 2010:101864. [PMID: 20182537 PMCID: PMC2825551 DOI: 10.1155/2010/101864] [Citation(s) in RCA: 60] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/25/2009] [Revised: 10/13/2009] [Accepted: 11/11/2009] [Indexed: 12/15/2022] Open
Abstract
There is a requirement for a noninvasive technique to monitor stem cell differentiation. Several candidates based on optical spectroscopy are discussed in this review: Fourier transform infrared (FTIR) spectroscopy, Raman spectroscopy, and coherent anti-Stokes Raman scattering (CARS) microscopy. These techniques are briefly described, and the ability of each to distinguish undifferentiated from differentiated cells is discussed. FTIR spectroscopy has demonstrated its ability to distinguish between stem cells and their derivatives. Raman spectroscopy shows a clear reduction in DNA and RNA concentrations during embryonic stem cell differentiation (agreeing with the well-known reduction in the nucleus to cytoplasm ratio) and also shows clear increases in mineral content during differentiation of mesenchymal stem cells. CARS microscopy can map these DNA, RNA, and mineral concentrations at high speed, and Mutliplex CARS spectroscopy/microscopy is highlighted as the technique with most promise for future applications.
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23
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Trevisan J, Angelov PP, Patel II, Najand GM, Cheung KT, Llabjani V, Pollock HM, Bruce SW, Pant K, Carmichael PL, Scott AD, Martin FL. Syrian hamster embryo (SHE) assay (pH 6.7) coupled with infrared spectroscopy and chemometrics towards toxicological assessment. Analyst 2010; 135:3266-72. [DOI: 10.1039/c0an00586j] [Citation(s) in RCA: 45] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/07/2023]
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Chan JW, Lieu DK. Label-free biochemical characterization of stem cells using vibrational spectroscopy. JOURNAL OF BIOPHOTONICS 2009; 2:656-668. [PMID: 19653219 DOI: 10.1002/jbio.200910041] [Citation(s) in RCA: 63] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/27/2023]
Abstract
Raman and infrared (IR) spectroscopy are two complementary vibrational spectroscopic techniques that have experienced a tremendous growth in their use in biological and biomedical research. This is, in large part, due to their unique capability of providing label-free intrinsic chemical information of living biological samples at tissue, cellular, or sub-cellular resolutions. This article reviews recent developments in applying these techniques for the characterization of stem cells. A discussion of the potential for these methods to address some of the major challenges in stem cell research is presented, as well as the technological and scientific advancements that are needed to progress the knowledge in the field.
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Affiliation(s)
- James W Chan
- NSF Center for Biophotonics Science and Technology, University of California, Davis, 2700 Stockton Blvd Suite 1400, Sacramento, CA 95817, USA.
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25
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Heraud P, Tobin MJ. The emergence of biospectroscopy in stem cell research. Stem Cell Res 2009; 3:12-4. [DOI: 10.1016/j.scr.2009.04.002] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/18/2009] [Accepted: 04/25/2009] [Indexed: 10/20/2022] Open
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26
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27
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Walsh MJ, Bruce SW, Pant K, Carmichael PL, Scott AD, Martin FL. Discrimination of a transformation phenotype in Syrian golden hamster embryo (SHE) cells using ATR-FTIR spectroscopy. Toxicology 2009; 258:33-8. [DOI: 10.1016/j.tox.2009.01.003] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/04/2008] [Accepted: 01/05/2009] [Indexed: 11/17/2022]
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28
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Walsh MJ, Hammiche A, Fellous TG, Nicholson JM, Cotte M, Susini J, Fullwood NJ, Martin-Hirsch PL, Alison MR, Martin FL. Tracking the cell hierarchy in the human intestine using biochemical signatures derived by mid-infrared microspectroscopy. Stem Cell Res 2009; 3:15-27. [PMID: 19393589 DOI: 10.1016/j.scr.2009.02.003] [Citation(s) in RCA: 47] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/16/2008] [Revised: 01/31/2009] [Accepted: 02/08/2009] [Indexed: 02/01/2023] Open
Abstract
Markers of gastrointestinal (GI) stem cells remain elusive. We employed synchrotron Fourier-transform infrared (FTIR) microspectroscopy to derive mid-infrared (IR) spectra along the length of human GI crypts. Tissue sections (10-μm thick) were floated onto BaF2 windows and image maps were acquired of small intestine and large bowel crypts in transmission mode with an aperture of ≤10 μm×10 μm. Counting upwards in a step-size (≤10 μm) fashion from the crypt base, IR spectra were extracted from the image maps and each spectrum corresponding to a particular location was identified. Spectra were analyzed using principal component analysis plus linear discriminant analysis. Compared to putative crypt base columnar/Paneth cells, those assigned as label-retaining cells were chemically more similar to putative large bowel stem cells and, the small intestine transit-amplifying cells were closest to large bowel transit-amplifying cells; interestingly, the base of small intestine crypts was the most chemically-distinct. This study suggests that in the complex cell lineage of human GI crypts, chemical similarities as revealed by FTIR microspectroscopy between regions putatively assigned as stem cell, transit-amplifying and terminally-differentiated facilitates identification of cell function.
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Affiliation(s)
- Michael J Walsh
- Centre for Biophotonics, Lancaster Environment Centre, Lancaster University, Bailrigg, Lancaster, UK
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29
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Kelly JG, Singh MN, Stringfellow HF, Walsh MJ, Nicholson JM, Bahrami F, Ashton KM, Pitt MA, Martin-Hirsch PL, Martin FL. Derivation of a subtype-specific biochemical signature of endometrial carcinoma using synchrotron-based Fourier-transform infrared microspectroscopy. Cancer Lett 2009; 274:208-17. [DOI: 10.1016/j.canlet.2008.09.018] [Citation(s) in RCA: 28] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/19/2008] [Revised: 07/19/2008] [Accepted: 09/10/2008] [Indexed: 11/16/2022]
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30
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Walsh MJ, Singh MN, Stringfellow HF, Pollock HM, Hammiche A, Grude O, Fullwood NJ, Pitt MA, Martin-Hirsch PL, Martin FL. FTIR Microspectroscopy Coupled with Two-Class Discrimination Segregates Markers Responsible for Inter- and Intra-Category Variance in Exfoliative Cervical Cytology. Biomark Insights 2008; 3:179-189. [PMID: 18677422 PMCID: PMC2493409 DOI: 10.4137/bmi.s592] [Citation(s) in RCA: 39] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/02/2022] Open
Abstract
Infrared (IR) absorbance of cellular biomolecules generates a vibrational spectrum, which can be exploited as a “biochemical fingerprint” of a particular cell type. Biomolecules absorb in the mid-IR (2–20 μm) and Fourier-transform infrared (FTIR) microspectroscopy applied to discriminate different cell types (exfoliative cervical cytology collected into buffered fixative solution) was evaluated. This consisted of cervical cytology free of atypia (i.e. normal; n = 60), specimens categorised as containing low-grade changes (i.e. CIN1 or LSIL; n = 60) and a further cohort designated as high-grade (CIN2/3 or HSIL; n = 60). IR spectral analysis was coupled with principal component analysis (PCA), with or without subsequent linear discriminant analysis (LDA), to determine if normal versus low-grade versus high-grade exfoliative cytology could be segregated. With increasing severity of atypia, decreases in absorbance intensity were observable throughout the 1,500 cm−1 to 1,100 cm−1 spectral region; this included proteins (1,460 cm−1), glycoproteins (1,380 cm−1), amide III (1,260 cm−1), asymmetric (νas) PO2− (1,225 cm−1) and carbohydrates (1,155 cm−1). In contrast, symmetric (νs) PO2− (1,080 cm−1) appeared to have an elevated intensity in high-grade cytology. Inter-category variance was associated with protein and DNA conformational changes whereas glycogen status strongly influenced intra-category. Multivariate data reduction of IR spectra using PCA with LDA maximises inter-category variance whilst reducing the influence of intra-class variation towards an objective approach to class cervical cytology based on a biochemical profile.
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Affiliation(s)
- Michael J Walsh
- Biomedical Sciences Unit, Department of Biological Sciences, Lancaster University, Lancaster, U.K
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31
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Ami D, Neri T, Natalello A, Mereghetti P, Doglia SM, Zanoni M, Zuccotti M, Garagna S, Redi CA. Embryonic stem cell differentiation studied by FT-IR spectroscopy. BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH 2008; 1783:98-106. [DOI: 10.1016/j.bbamcr.2007.08.003] [Citation(s) in RCA: 81] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/11/2007] [Revised: 08/01/2007] [Accepted: 08/02/2007] [Indexed: 11/28/2022]
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32
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Walsh MJ, Fellous TG, Hammiche A, Lin WR, Fullwood NJ, Grude O, Bahrami F, Nicholson JM, Cotte M, Susini J, Pollock HM, Brittan M, Martin-Hirsch PL, Alison MR, Martin FL. Fourier transform infrared microspectroscopy identifies symmetric PO(2)(-) modifications as a marker of the putative stem cell region of human intestinal crypts. Stem Cells 2008; 26:108-118. [PMID: 17901405 DOI: 10.1634/stemcells.2007-0196] [Citation(s) in RCA: 93] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/17/2023]
Abstract
Complex biomolecules absorb in the mid-infrared (lambda = 2-20 microm), giving vibrational spectra associated with structure and function. We used Fourier transform infrared (FTIR) microspectroscopy to "fingerprint" locations along the length of human small and large intestinal crypts. Paraffin-embedded slices of normal human gut were sectioned (10 microm thick) and mounted to facilitate infrared (IR) spectral analyses. IR spectra were collected using globar (15 microm x 15 microm aperture) FTIR microspectroscopy in reflection mode, synchrotron (
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Affiliation(s)
- Michael J Walsh
- Biomedical Sciences Unit, Department of Biological Sciences, Lancaster University, Bailrigg, Lancaster LA1 4YQ, UK
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GRUDE OLAUG, HAMMICHE AZZEDINE, POLLOCK HUBERT, BENTLEY ADAMJ, WALSH MICHAELJ, MARTIN FRANCISL, FULLWOOD NIGELJ. Near-field photothermal microspectroscopy for adult stem-cell identification and characterization. J Microsc 2007; 228:366-72. [DOI: 10.1111/j.1365-2818.2007.01853.x] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/28/2022]
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Remington SG, Meyer RA. Lens stem cells may reside outside the lens capsule: an hypothesis. Theor Biol Med Model 2007; 4:22. [PMID: 17559656 PMCID: PMC1914343 DOI: 10.1186/1742-4682-4-22] [Citation(s) in RCA: 14] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/18/2006] [Accepted: 06/08/2007] [Indexed: 01/21/2023] Open
Abstract
In this paper, we consider the ocular lens in the context of contemporary developments in biological ideas. We attempt to reconcile lens biology with stem cell concepts and a dearth of lens tumors. Historically, the lens has been viewed as a closed system, in which cells at the periphery of the lens epithelium differentiate into fiber cells. Theoretical considerations led us to question whether the intracapsular lens is indeed self-contained. Since stem cells generate tumors and the lens does not naturally develop tumors, we reasoned that lens stem cells may not be present within the capsule. We hypothesize that lens stem cells reside outside the lens capsule, in the nearby ciliary body. Our ideas challenge the existing lens biology paradigm. We begin our discussion with lens background information, in order to describe our lens stem cell hypothesis in the context of published data. Then we present the ciliary body as a possible source for lens stem cells, and conclude by comparing the ocular lens with the corneal epithelium.
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Affiliation(s)
- Susann G Remington
- Ophthalmology Research, HealthPartners Medical Group and Research Foundation, Regions Hospital, 640 Jackson Street, St. Paul, MN 55101, USA
| | - Rita A Meyer
- Department of Biomedical Sciences, Creighton University, Criss I, Room 217, 2500 California Plaza, Omaha, NE 68178, USA
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Walsh MJ, German MJ, Singh M, Pollock HM, Hammiche A, Kyrgiou M, Stringfellow HF, Paraskevaidis E, Martin-Hirsch PL, Martin FL. IR microspectroscopy: potential applications in cervical cancer screening. Cancer Lett 2007; 246:1-11. [PMID: 16713674 DOI: 10.1016/j.canlet.2006.03.019] [Citation(s) in RCA: 103] [Impact Index Per Article: 5.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/15/2006] [Revised: 03/14/2006] [Accepted: 03/16/2006] [Indexed: 01/11/2023]
Abstract
Screening exfoliative cytology for early dysplastic cells reduces incidence and mortality from squamous carcinoma of the cervix. In the developed world, screening programmes have adopted a 3-5 years recall system. In its absence, cervical cancer would be the second most common female cancer in these regions; instead, it is currently eleventh. However, there exist a number of limitations to the smear test even given the removal of contaminants using liquid-based cytology. It is prohibitively expensive, labour-intensive and subject to inaccuracies that give rise to significant numbers of false negatives. There remains a need for novel approaches to allow efficient and objective interrogation of exfoliative cytology. Methods that variously exploit infrared (IR) microspectroscopy are one possibility. Using IR microspectroscopy, an integrated 'biochemical-cell fingerprint' of the lipid, protein and carbohydrate composition of a biomolecular entity may be derived in the form of a spectrum via vibrational transitions of individual chemical bonds. Powerful statistical approaches (e.g. principal component analysis) now facilitate the interrogation of large amounts of spectroscopic data to allow the extraction of what may be small but extremely significant biomarker differences between disease-free and pre-malignant or malignant samples. An increasing wealth of literature points to the ability of IR microspectroscopy to allow the segregation of cells based on their disease status. We review the current evidence supporting its diagnostic potential in cancer biology.
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Affiliation(s)
- Michael J Walsh
- Biomedical Sciences Unit, Department of Biological Sciences, Lancaster University, Lancaster, UK
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Hammiche A, Walsh MJ, Pollock HM, Martin-Hirsch PL, Martin FL. Non-contact micro-cantilevers detect photothermally induced vibrations that can segregate different categories of exfoliative cervical cytology. ACTA ACUST UNITED AC 2007; 70:675-7. [PMID: 17320188 DOI: 10.1016/j.jbbm.2007.01.011] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/24/2006] [Accepted: 01/21/2007] [Indexed: 11/29/2022]
Abstract
We implemented a non-contact photo-thermo-mechanical recording method whereby a silicon nitride atomic force microscopy cantilever is placed several micrometer above the surface of samples. Samples were illuminated with infrared (IR) radiation after which, cantilever mechanical vibrations were optically sensed. Following spectrometric acquisition and Fourier transformation, true IR absorption spectra were obtained. With multivariate analysis, segregation between different categories of exfoliative cervical cytology was obtained. This approach points towards the implementation of a novel near-field system that allows IR spectral analysis without probe contamination.
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Dumas P, Sockalingum GD, Sulé-Suso J. Adding synchrotron radiation to infrared microspectroscopy: what's new in biomedical applications? Trends Biotechnol 2007; 25:40-4. [PMID: 17116340 DOI: 10.1016/j.tibtech.2006.11.002] [Citation(s) in RCA: 96] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/27/2006] [Revised: 10/06/2006] [Accepted: 11/07/2006] [Indexed: 10/23/2022]
Abstract
Infrared spectroscopy and microscopy have heralded a period of rapid advances in tissue and cellular characterization during the past decade. However, vibrational spectroscopy is still an analytical tool that is neither familiar nor understood in the medical environment. For many years this field has been mainly driven by physicists and chemists, who are, undoubtedly, at the forefront of tremendous technical developments in technology, detection and data treatment. Although the theory of infrared (IR) spectroscopy is thoroughly worked out, the scientific ground of vibrational spectroscopy is now undergoing a real boost, with the application of this analytical technique in biology and biomedicine.
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Affiliation(s)
- Paul Dumas
- SOLEIL Synchrotron, L'orme des Merisiers, BP48, F-91191 Gif sur Yvette Cédex, France.
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Wood BR, Bambery KR, Evans CJ, Quinn MA, McNaughton D. A three-dimensional multivariate image processing technique for the analysis of FTIR spectroscopic images of multiple tissue sections. BMC Med Imaging 2006; 6:12. [PMID: 17014733 PMCID: PMC1592472 DOI: 10.1186/1471-2342-6-12] [Citation(s) in RCA: 43] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/11/2006] [Accepted: 10/03/2006] [Indexed: 11/16/2022] Open
Abstract
Background Three-dimensional (3D) multivariate Fourier Transform Infrared (FTIR) image maps of tissue sections are presented. A villoglandular adenocarcinoma from a cervical biopsy with a number of interesting anatomical features was used as a model system to demonstrate the efficacy of the technique. Methods Four FTIR images recorded using a focal plane array detector of adjacent tissue sections were stitched together using a MATLAB® routine and placed in a single data matrix for multivariate analysis using Cytospec™. Unsupervised Hierarchical Cluster Analysis (UHCA) was performed simultaneously on all 4 sections and 4 clusters plotted. The four UHCA maps were then stacked together and interpolated with a box function using SCIRun software. Results The resultant 3D-images can be rotated in three-dimensions, sliced and made semi-transparent to view the internal structure of the tissue block. A number of anatomical and histopathological features including connective tissue, red blood cells, inflammatory exudate and glandular cells could be identified in the cluster maps and correlated with Hematoxylin & Eosin stained sections. The mean extracted spectra from individual clusters provide macromolecular information on tissue components. Conclusion 3D-multivariate imaging provides a new avenue to study the shape and penetration of important anatomical and histopathological features based on the underlying macromolecular chemistry and therefore has clear potential in biology and medicine.
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Affiliation(s)
- Bayden R Wood
- Centre for Biospectroscopy and School of Chemistry, Monash University, 3800 Victoria, Australia
| | - Keith R Bambery
- Centre for Biospectroscopy and School of Chemistry, Monash University, 3800 Victoria, Australia
| | - Corey J Evans
- Department of Chemistry, University of Leicester, Leicester, LE1 7RH, UK
| | - Michael A Quinn
- Department of Obstetrics and Gynaecology, Royal Women's Hospital, Grattan St. Parkville, 3052, Victoria, Australia Sciences, Monash University, 3800 Victoria, Australia
| | - Don McNaughton
- Centre for Biospectroscopy and School of Chemistry, Monash University, 3800 Victoria, Australia
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