The Molecular Phenotyping core provides instrumentation, training, and services for in vitro cell phenotyping. Specifically, we provide facilities and training for automated nucleic acids purification, microarray analysis, quantitative PCR, and automated immunofluorescence cell marker analysis and screening. In addition to maintaining facilities essential for the many NIH-funded investigators working at MMCRI, planned developments such as resources for analysis of next generation sequencing data and laboratory automation for high density microplate formats will ensure that the core keeps in step with investigators'needs. An efficient workflow for the analysis of gene expression from RNA preparation through transcriptome analysis, and validation of candidate genes has been established. The core facility provides equipment, training, and supplies for all steps in the process with the exception of microarray processing, which is provided by Vermont Genetics Network in a regional collaborative agreement. Both manual and automated epifluorescence microscopy is also provided by the facility. Currently we provide live cell time-lapse microscopy, and microscopy based screening in 6- to 96-well formats, and additional applications such as cell cycle analysis are in development. The core provides image analysis workstations running Metamorph and Image J, and training in specific applications of these software packages. Quantitative image analysis of tissue immunostaining is being provided in collaboration with the Histopathology Core (Lindner). Future developments of our transcriptome analysis capabilities will be in the following areas: i) Transition from 96- to 384-well format. Essential equipment including 384 well real-time PCR instrument and liquid handling station is available in the facility, and users are currently piloting the new platform, ii) Providing an analysis platform for Next Generation Sequencing data. In the cellular analysis area we will be focusing on increasing the number of analyses we can provide users of our automated microscopy platform. Furthermore, we will be transitioning to a 384 well format for microscopy-based screening applications. Longer term core development will be based on feedback from the user base, and input from the internal and external advisory committees of the COBRE.

Public Health Relevance

State of the art resources for transcriptome analysis and cellular marker analysis are essential to nationally competitive biomedical research infrastructure. The Molecular Phenotyping core has developed these resources at MMCRI, and continues to add new resources in these areas on the basis of the needs of MMCRI investigators.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Center Core Grants (P30)
Project #
1P30GM106391-01
Application #
8543893
Study Section
Special Emphasis Panel (ZGM1-TWD-C (C3))
Project Start
Project End
Budget Start
2013-09-01
Budget End
2014-05-31
Support Year
1
Fiscal Year
2013
Total Cost
$70,432
Indirect Cost
$24,548
Name
Maine Medical Center
Department
Type
DUNS #
071732663
City
Portland
State
ME
Country
United States
Zip Code
04102
Ames, Jacquelyn J; Contois, Liangru; Caron, Jennifer M et al. (2016) Identification of an Endogenously Generated Cryptic Collagen Epitope (XL313) That May Selectively Regulate Angiogenesis by an Integrin Yes-associated Protein (YAP) Mechano-transduction Pathway. J Biol Chem 291:2731-50
Liaw, Lucy; Prudovsky, Igor; Koza, Robert A et al. (2016) Lipid Profiling of In Vitro Cell Models of Adipogenic Differentiation: Relationships With Mouse Adipose Tissues. J Cell Biochem 117:2182-93
Dadwal, Ushashi; Falank, Carolyne; Fairfield, Heather et al. (2016) Tissue-engineered 3D cancer-in-bone modeling: silk and PUR protocols. Bonekey Rep 5:842
Anunciado-Koza, Rea P; Higgins, David C; Koza, Robert A (2016) Adipose tissue Mest and Sfrp5 are concomitant with variations of adiposity among inbred mouse strains fed a non-obesogenic diet. Biochimie 124:134-40
Lecka-Czernik, Beata; Rosen, Clifford J (2016) Skeletal integration of energy homeostasis: Translational implications. Bone 82:35-41
Krebs, Luke T; Norton, Christine R; Gridley, Thomas (2016) Notch signal reception is required in vascular smooth muscle cells for ductus arteriosus closure. Genesis 54:86-90
Young, K; Krebs, L T; Tweedie, E et al. (2016) Endoglin is required in Pax3-derived cells for embryonic blood vessel formation. Dev Biol 409:95-105
Calabrese, Gina; Mesner, Larry D; Foley, Patricia L et al. (2016) Network Analysis Implicates Alpha-Synuclein (Snca) in the Regulation of Ovariectomy-Induced Bone Loss. Sci Rep 6:29475
Martinez, M Elena; Karaczyn, Aldona; Stohn, J Patrizia et al. (2016) The Type 3 Deiodinase Is a Critical Determinant of Appropriate Thyroid Hormone Action in the Developing Testis. Endocrinology 157:1276-88
Anunciado-Koza, Rea P; Manuel, Justin; Koza, Robert A (2016) Molecular correlates of fat mass expansion in C57BL/6J mice after short-term exposure to dietary fat. Ann N Y Acad Sci 1363:50-8

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