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High-Throughput 3D Cell Invasion Chip Enables Accurate Cancer Metastatic Assays

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 2014年第43期136卷 15257-15262页

作者： Zhang, Yuanqing Zhou, Ledu Qin, Lidong Houston Methodist Res Inst Dept Nanomed Houston TX 77030 USA Cornell Univ Weill Med Coll Dept Cell & Dev Biol New York NY 10065 USA Univ Texas MD Anderson Canc Ctr Dept Mol & Cellular Oncol Houston TX 77030 USA Cent S Univ Xiangya Hosp Dept Gen Surg Changsha 410008 Hunan Peoples R China

Chemotaxis is the phenomenon by which the migration and invasion of cells is directed in response to an extracellular chemical gradient. Chemotaxis of tumor cells and tumor-associated inflammatory and stromal cells is mediated by chemokines, chemokine receptors, growth factors, and growth factor receptors. Current techniques used to study chemotactic driven cell invasion and metastasis utilize two-dimensional migration assays involving imaging and analyzing tumor cells on glass slides or plastic surfaces, which requires large numbers of cells and often lacks real-time monitoring of vertical cell movement and systematically controlled chemotactic gradients, leading to contradictory results compared to those from clinical investigations and animal models. We addressed such challenges by developing a high-throughput microdevice with 4000 ultraminiaturized wells to monitor real-time, three-dimensional cell invasion over a wide range of cell densities and also screen drugs that inhibit cell invasion and potentially prevent metastatic malignancy. Additionally, this microdevice generates opposing gradients for two types of cells on the same chip, which builds a controlled system with sequentially changing components to study environmental effects from basal and immune cells.

关键词： Cell Count Cell Count Chemotaxis Collagen/chemistry Humans Macrophages/pathology MCF-7 Cells Neoplasm Invasiveness Neoplasm Metastasis Neoplasms/pathology Single-Cell analysis tissue array analysis/methods

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Advanced Substrate Fabrication for Cell Microarrays

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BIOMACROMOLECULES 2009年第3期10卷 573-579页

作者： Hook, Andrew L. Thissen, Helmut Voelcker, Nicolas H. Flinders Univ S Australia Sch Chem Phys & Earth Sci Bedford Pk SA 5042 Australia CSIRO Mol & Hlth Technol Clayton Vic 3168 Australia

The fabrication and characterization of chemical patterns using a technique that can be readily integrated with methods currently used for the formation of microarrays is presented. A high density poly(ethylene glycol) coating was deposited on glass slides as a background exhibiting low cell attachment properties. Phenylazide modified polymers were then printed on this background. UV irradiation of these polymer arrays resulted in the cross-linking of the polymer spots and their covalent attachment to the surface. Cell attachment was shown to follow the resultant surface chemistry pattern. Furthermore, the use of a robotic contact printer enabled the facile deposition of DNA microarrays on top of and aligned with the polymer microarrays. A transfected cell microarray was generated in this way, demonstrating not only the ability of this platform to limit cell attachment to specific regions, but the suitability for chip-based functional genomics, in particular, and high density cell assays in general.

关键词： Azides/chemistry Cell Proliferation Coated Materials, Biocompatible/chemical synthesis Coated Materials, Biocompatible/chemistry Glass/chemistry Humans Macromolecular Substances/chemical synthesis Macromolecular Substances/chemistry Particle Size Polyethylene Glycols/chemistry Surface Properties tissue array analysis/methods Tumor Cells, Cultured Ultraviolet Rays Ultraviolet Rays

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Matrix-Assisted Laser Desorption Ionization Imaging Mass Spectrometry: In Situ Molecular Mapping

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BIOCHEMISTRY 2013年第22期52卷 3818-3828页

作者： Angel, Peggi M. Caprioli, Richard M. Vanderbilt Univ Med Ctr Mass Spectrometry Res Ctr Nashville TN 37232 USA Vanderbilt Univ Med Ctr Dept Biochem Nashville TN 37232 USA Vanderbilt Univ Med Ctr Dept Biochem Med Pharmacol & Chem Nashville TN 37232 USA

Matrix-assisted laser desorption ionization imaging mass spectrometry (IMS) is a relatively new imaging modality that allows Mapping of a wide range of biomolecules within a thin tissue section. The technology uses a laser beam to directly desorb and ionize molecules from discrete locations on the tissue that are subsequently recorded in a mass spectrometer. IMS is distinguished by the ability to directly measure molecules in situ ranging from small, metabolites to proteins, reporting hundreds to thousands of expression patterns from a single imaging experiment. This article reviews recent advances in IMS technology, applications, and experimental strategies that allow it to significantly aid in the discovery and understanding of molecular processes in biological and clinical samples.

关键词： Animals Breast Neoplasms/chemistry Embryo, Mammalian/chemistry Fixatives Lung/chemistry Mice Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/instrumentation Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods tissue array analysis/methods

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tissue microarray construction from bone marrow biopsies

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BIOTECHNIQUES 2005年第6期39卷 822-+页

作者： Obermann, EC Marienhagen, J Stoehr, R Wuensch, PH Hofstaedter, F Univ Regensburg Inst Pathol D-93053 Regensburg Germany Clin Ctr Nuremberg Nurnberg Germany

No abstract available

关键词： Biopsy Biopsy Bone Marrow/pathology Humans tissue array analysis/methods

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Surface Engineering for Long-Term Culturing of Mesenchymal Stem Cell Microarrays

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BIOMACROMOLECULES 2013年第8期14卷 2675-2683页

作者： Ghaemi, Soraya Rasi Harding, Frances Delalat, Bahman Vasani, Roshan Voelcker, Nicolas H. Univ S Australia Mawson Inst Adelaide SA 5095 Australia

The cell microarray format can recreate a multitude of cell microenvironments on a single chip using only minimal amounts of reagent. In this study, we describe surface modifications to passivate cell microarrays, aiming to adapt the platform to the study of stem cell behavior over long-term culture periods. Functionalization of glass slides with (3-glycidyloxypropyl) trimethoxysilane enabled covalent anchoring of extracellular matrix proteins on microscale spots printed by a robotic contact printer. Subsequently, the surface was passivated by bovine serum albumin (BSA) or poly(ethylene glycol)bisamine (A-PEG) with molecular weights of 3000, 6000, and 10 000 Da. Cloud-point conditions for A-PEG grafting were attained that were compatible with protein deposition. Passivation strategies were assessed by culturing mesenchymal stem cells on the microarray platform. While both BSA and A-PEG passivation initially blocked cell adhesion between the printed spots, only A-PEG grafting was able to maintain cell pattern integrity over the entire culture period of 3 weeks.

关键词： Animals Cell Adhesion Cell Culture Techniques Cell Culture Techniques Cell Differentiation Cell Survival Extracellular Matrix Proteins/chemistry Humans Immobilized Proteins/chemistry Jurkat Cells Mesenchymal Stromal Cells/physiology Polyethylene Glycols/chemistry Rats Rats, Wistar Sulfates/chemistry Surface Properties tissue array analysis/methods

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Sensitive and high resolution subcutaneous fluorescence in vivo imaging using upconversion nanoparticles and microarrays

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ANALYST 2013年第13期138卷 3711-3718页

作者： Li, Xin Li, Zhuoqi Gan, Wupeng Wang, Tongzhou Zhao, Songmin Lu, Ying Cheng, Jing Huang, Guoliang Tsinghua Univ Sch Med Dept Biomed Engn Beijing 100084 Peoples R China Natl Engn Res Ctr Beijing Biochip Technol Beijing 102206 Peoples R China

A sensitive and high resolution small animal in vivo imaging system using upconversion nanoparticles (UNPs) and microarrays was developed. The fluorescence tomography using UNPs could achieve higher precision than that using ordinary fluorophores, which was theoretically explained by the finite element method (FEM). Given the autofluorescence-insensitive property of UNPs, a high subcutaneous detection sensitivity of 0.93 x 10(-4) wt% could be achieved with a UNP volume of similar to 10 mu L in tissue phantoms. Furthermore, UNP fluorophore microarrays (25, 50 and 100 mm arrays) embedded under mouse skin were prepared for subcutaneous in vivo detection. An optical clearing method was applied to enhance the skin transparency and improve the spatial resolution. The results demonstrated that the optimized system could achieve a spatial resolution of 50 mm for in vivo detection of subcutaneous UNP microarrays. Taken together, we conclude that the proposed system and UNP microarrays could achieve sensitive, high resolution subcutaneous in vivo detection, and have great potential for high throughput detection of tumors and other diseases.

关键词： Animals Mice Mice, Inbred BALB C Molecular Imaging/methods Nanoparticles/chemistry Skin/cytology Spectrometry, Fluorescence Subcutaneous tissue/metabolism tissue array analysis/methods

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tissue analysis by imaging MS

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BIOanalysis 2012年第1期4卷 95-112页

作者： Oppenheimer, Stacey R. Drexler, Dieter M. Pfizer Global Res & Dev World Wide Med Chem Groton CT 06340 USA Bristol Myers Squibb Co Res & Dev Discovery Analyt Sci Wallingford CT 06492 USA

Imaging MS (IMS) is generating tremendous interest in scientific communities because of its unparalleled capabilities to provide chemical analysis of intact tissue. Advances in analytical chemistry and MS are providing new insights into chemical and biological processes. This review will discuss various IMS platforms and their applications in biomedical and pharmaceutical research.

关键词： Animals Humans Image Processing, Computer-Assisted/methods Mass Spectrometry/methods tissue array analysis/methods tissue Distribution

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Label-free detection of surface markers on stem cells by oblique-incidence reflectivity difference microscopy

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BIOTECHNIQUES 2011年第6期50卷 381-+页

作者： Lo, Kai-Yin Sun, Yung-Shin Landry, James P. Zhu, Xiangdong Deng, Wenbin Univ Calif Davis Dept Cell Biol & Human Anat Inst Pediat Regenerat Med Sch Med Davis CA 95616 USA Univ Calif Davis Dept Phys Davis CA 95616 USA Natl Taiwan Univ Dept Agr Chem Taipei 10764 Taiwan

Conventional fluorescence microscopy is routinely used to detect cell surface markers through fluorophore-conjugated antibodies. However, fluorophore-conjugation of antibodies alters binding properties such as strength and specificity of the antibody in often uncharacterized ways. Here we present a method using an oblique-incidence reflectivity difference (OI-RD) microscope for label-free, real-time detection of cell surface markers, and apply it to analysis of stage-specific embryonic antigen 1 (SSEA1) on stem cells. Mouse stem cells express SSEA1 on their surfaces, and the level of SSEA1 decreases when the cells start to differentiate. In this study, we immobilized mouse stem cells and non stem cells (control) on a glass surface as a microarray and reacted the cell microarray with unlabeled SSEA1 antibodies. By monitoring the reaction with an OI-RD microscope in real time, we confirmed that the SSEA1 antibodies bind only to the surface of the stem cells and not to the surface of non stem cells. From the binding curves, we determined the equilibrium dissociation constant (K(d)) of the antibody with the SSEA1 markers on the stem cell surface. Thus, the OI-RD microscope can be used to detect binding affinities between cell surface markers and unlabeled antibodies bound to the cells;this information could be useful for determination of stem cell stages.

关键词： Animals Antibodies/metabolism Antigens, CD15/chemistry Antigens, CD15/metabolism Biological Markers/chemistry Cell Differentiation Cells, Immobilized/cytology Cells, Immobilized/metabolism Cytological Techniques Cytological Techniques Mice Microscopy/methods Microscopy, Fluorescence Protein Binding Stem Cells/chemistry Stem Cells/cytology Stem Cells/metabolism tissue array analysis/methods

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2011 White Paper on Recent Issues in Bioanalysis and Regulatory Findings from Audits and Inspections

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BIOanalysis 2011年第18期3卷 2081-2096页

作者： Garofolo, Fabio Rocci, Mario L., Jr. Dumont, Isabelle Martinez, Suzanne Lowes, Steve Woolf, Eric van Amsterdam, Peter Bansal, Surendra Gomes Barra, Ariadna Cristina Bauer, Ronald Booth, Brian P. Carrasco-Triguero, Montserrat DeSilva, Binodh Dunn, John Gallicano, Keith Gouty, Dominique Ho, Stacy Hucker, Richard Jemal, Mohammed Katori, Noriko Le Blaye, Olivier Lee, Jean Li, Wenkui Michael, Steve Nehls, Corey Nicholson, Robert Ormsby, Eric Tang, Daniel Viswanathan, C. T. Weiner, Russell Young, Graeme Algorithme Pharma Inc Montreal PQ H7V 4B3 Canada ICON Dev Solut Whitesboro NY USA Advion BioServ Inc Ithaca NY USA Merck Res Labs West Point PA USA Abbott Healthcare Prod BV Weesp Netherlands Hoffmann La Roche Nutley NJ USA ANVISA Brasilia DF Brazil AGES PharmMed Vienna Austria US FDA CDER Silver Spring MD USA Genentech Inc San Francisco CA 94080 USA Bristol Myers Squibb Co Princeton NJ USA GlaxoSmithKline Res & Dev Ltd Res Triangle Pk NC USA Watson Labs Corona CA USA Intertek San Diego CA USA Genzyme Corp Waltham MA USA Pfizer Sandwich Kent England Japan MHLW NIHS Tokyo Japan AFSSAPS St Denis France Amgen Inc Thousand Oaks CA 91320 USA Novartis Inst Biomed Res E Hanover NJ USA Covance Madison WI USA PPD Middleton WI USA PPD Richmond VA USA Hlth Canada TPD Ottawa ON Canada Frontage Labs Shanghai Peoples R China Merck Res Labs Rahway NJ USA GlaxoSmithKline Res & Dev Ltd Ware Herts England

The 5th Workshop on Recent Issues in Bioanalysis (WRIB) was organized by the Calibration and Validation Group as a 2-day full immersion workshop for pharmaceutical companies, CROs and regulatory agencies to discuss, review, share perspectives, provide potential solutions and agree upon a consistent approach to recent issues in the bioanalysis of both small and large molecules. High quality, better compliance to regulations and scientific excellence are the foundation of this workshop. As in the previous editions of this significant event, recommendations were made and a consensus was reached among panelists and attendees, including industry leaders and regulatory experts representing the global bioanalytical community, on many 'hot' topics in bioanalysis. This 2011 White Paper is based on the conclusions from this workshop, and aims to provide a practical reference guide on those topics.

关键词： Calibration Chromatography, High Pressure Liquid/methods Chromatography, High Pressure Liquid/standards Dried Blood Spot Testing/methods Dried Blood Spot Testing/standards Drug Industry Government Regulation Guidelines as Topic Humans Mass Spectrometry/instrumentation Mass Spectrometry/methods Mass Spectrometry/standards Pharmaceutical Preparations/analysis Technology Transfer tissue array analysis/methods tissue array analysis/standards Validation Studies as Topic

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TTF-1 expression in primary ovarian epithelial neoplasia

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HISTOPATHOLOGY 2006年第6期48卷 764-765页

作者： Graham, AD Williams, ARW Salter, DM Royal Infirm Dept Histopathol Edinburgh Midlothian Scotland

A letter to the editor is presented about thyroid transcription factor-1 in primary ovarian epithelial neoplasia.

关键词： Adenocarcinoma, Clear Cell/metabolism Adenocarcinoma, Clear Cell/pathology Carcinoma, Endometrioid/metabolism Carcinoma, Endometrioid/pathology Cystadenocarcinoma, Mucinous/metabolism Cystadenocarcinoma, Mucinous/pathology Cystadenocarcinoma, Serous/metabolism Cystadenocarcinoma, Serous/pathology Epithelial Cells/chemistry Epithelial Cells/pathology Female Humans Immunohistochemistry Immunohistochemistry Neoplasm Staging Neoplasm Staging Nuclear Proteins/biosynthesis Ovarian Neoplasms/metabolism Ovarian Neoplasms/pathology tissue array analysis/methods Transcription Factors/biosynthesis

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