Abstract

OVERVIEW The Leica DM-RXA is a fully computer-controlled light microscope. The DAR is collaborating with Dr. Mueller's lab to design and test a software interface which will allow 3D time-lapse (4D) studies to be conducted using this microscope. The bulk of this project will involve Mr. Thomas writing, testing and debugging computer code as well as doing real-world testing of data collection using the Leica DM-RXA microscope itself. Dr. Fleischmann will then be involved in testing and debugging the software as well as in pursuing real-world biological applications of the software in his lab in Switzerland. This project is divided into two parts. STAGE 1: We intend to develog both a C-based stand-alone and an NIH-Image dependent software interface to allow the DM-RXA microscope to function in the collection of basic 4-D data sets using high-resolution Nomarski DIC optics. This includes the orchestration of illumination shutters, stage motors and digitizing video fi-amegrabbers from a Macintosh PPC computer as outlined in Thomas et al., (1996) Science, 273, 603-607. Easy-to-use and intuitive graphic interfaces should allow this software to be uncomplicated and permit a low learning curve for researchers hoping to become involved in 4D research. The software should also be made available free of charge via the internet to researchers around the world. NOTE This phase of the project has been completed successfully. STAGE 2: This phase of the project will investigate the possibility of gathering simultaneous (or nearly simultaneous) DIC and fluorescent 4D data sets from the same sample. BIOLOGICAL RESEARCH PLANS: The biological research plans for each phase of this project are as follows: Stage I - determination of the lineage of newly isolated mutant embryos. Stage 2: Expression studies with strains carrying GFP reporter constructs. Our main focus lies on the ceh-13 gene, but it could be used for any gene. First we will determine the wild type expression pattern. Then we will analyze mutants (which we think differ from the wild VyW expression pattern) and strains carrying deletions in the promoter region of the GFP reporter construct.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
3P41RR000570-28
Application #
6278469
Study Section
Project Start
1998-07-01
Project End
2000-06-30
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
28
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
University of Wisconsin Madison
Department
Type
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715

  Publications

Malecki, Marek; Putzer, Emily; Sabo, Chelsea et al. (2014) Directed cardiomyogenesis of autologous human induced pluripotent stem cells recruited to infarcted myocardium with bioengineered antibodies. Mol Cell Ther 2:
Malecki, Marek (2014) 'Above all, do no harm': safeguarding pluripotent stem cell therapy against iatrogenic tumorigenesis. Stem Cell Res Ther 5:73
Mavroudi, Maria; Zarogoulidis, Paul; Porpodis, Konstantinos et al. (2014) Stem cells' guided gene therapy of cancer: New frontier in personalized and targeted therapy. J Cancer Res Ther (Manch) 2:22-33
Malecki, Marek; LaVanne, Christine; Alhambra, Dominique et al. (2013) Safeguarding Stem Cell-Based Regenerative Therapy against Iatrogenic Cancerogenesis: Transgenic Expression of DNASE1, DNASE1L3, DNASE2, DFFB Controlled By POLA1 Promoter in Proliferating and Directed Differentiation Resisting Human Autologous Pluripotent J Stem Cell Res Ther Suppl 9:
Malecki, Marek; Tombokan, Xenia; Anderson, Mark et al. (2013) TRA-1-60(+), SSEA-4(+), POU5F1(+), SOX2(+), NANOG(+) Clones of Pluripotent Stem Cells in the Embryonal Carcinomas of the Testes. J Stem Cell Res Ther 3:
Malecki, Marek (2013) Improved targeting and enhanced retention of the human, autologous, fibroblast-derived, induced, pluripotent stem cells to the sarcomeres of the infarcted myocardium with the aid of the bioengineered, heterospecific, tetravalent antibodies. J Stem Cell Res Ther 3:
Malecki, Marek; Dahlke, Jessica; Haig, Melissa et al. (2013) Eradication of Human Ovarian Cancer Cells by Transgenic Expression of Recombinant DNASE1, DNASE1L3, DNASE2, and DFFB Controlled by EGFR Promoter: Novel Strategy for Targeted Therapy of Cancer. J Genet Syndr Gene Ther 4:152
Zarogoulidis, Paul; Darwiche, Kaid; Sakkas, Antonios et al. (2013) Suicide Gene Therapy for Cancer - Current Strategies. J Genet Syndr Gene Ther 4:
Malecki, Marek; Sabo, Chelsea; Putzer, Emily et al. (2013) Recruitment and retention of human autologous CD34+ CD117+ CD133+ bone marrow stem cells to infarcted myocardium followed by directed vasculogenesis: Novel strategy for cardiac regeneration. Mol Cell Ther 1:
Malecki, Marek; Malecki, Bianca (2012) Nuclear routing networks span between nuclear pore complexes and genomic DNA to guide nucleoplasmic trafficking of biomolecules. J Fertili In Vitro 2:

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