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Fred Hutchinson Cancer Research Center - FHCRC

5 ARTICLES PUBLISHED IN JoVE

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Biology

Generating Chimeric Zebrafish Embryos by Transplantation
Hilary A. Kemp 1, Amanda Carmany-Rampey 1, Cecilia Moens 1
1HHMI and Division of Basic Sciences, Fred Hutchinson Cancer Research Center - FHCRC

A step-by-step guide to generating targeted chimeric zebrafish embryos by transplantation at the blastula or gastrula stage.

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Biology

A High-Throughput Method For Zebrafish Sperm Cryopreservation and In Vitro Fertilization
Bruce W. Draper 1, Cecilia B. Moens 2
1Molecular and Cellular Biology, University of California, Davis, 2Division of Basic Science, Fred Hutchinson Cancer Research Center - FHCRC

This is a high-throughput sperm cryopreservation protocol for zebrafish. Sperm cryopreserved using this protocol has an average of 25% fertility in subsequent vitro fertilization and is stable over many years.

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Biology

Making Gynogenetic Diploid Zebrafish by Early Pressure
Charline Walker 1, Greg S. Walsh 2, Cecilia Moens 2
1Institute of Neuroscience, University of Oregon, 2Division of Basic Science, Fred Hutchinson Cancer Research Center - FHCRC

This is a method for generating gynogenetic diploid zebrafish embryos (embryos whose only genetic contribution comes from the mother) by blocking the second meiotic division immediately after fertilization with ultraviolet light-inactivated sperm. EP embryos are not fully homozygous due to recombination during the first meiotic division, however they are homozygous at all loci that have not been separated from their centromere by recombination.

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Biology

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
Lei Zhao *1, Jeffrey R. Whiteaker *1, Matthew E. Pope 2, Eric Kuhn 3, Angela Jackson 4, N. Leigh Anderson 5, Terry W. Pearson 2, Steven A. Carr 3, Amanda G. Paulovich 1
1Clinical Research Division, Fred Hutchinson Cancer Research Center - FHCRC, 2Department of Biochemistry and Microbiology, University of Victoria, 3Broad Institute of MIT and Harvard, 4Genome BC Proteomics Centre, University of Victoria, 5Plasma Proteome Institute

Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA) couples affinity enrichment of peptides with stable isotope dilution mass spectrometry (MRM-MS) to provide quantitative measurement of peptides as surrogates for their respective proteins. Here we describe the protocol using magnetic particles in a partially automated format.

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Behavior

Brain Infarct Segmentation and Registration on MRI or CT for Lesion-symptom Mapping
J. Matthijs Biesbroek 1, Hugo J. Kuijf 2, Nick A. Weaver 1, Lei Zhao 3, Marco Duering 4, , Geert Jan Biessels 1
1Department of Neurology and Neurosurgery, UMC Utrecht Brain Center, University Medical Center Utrecht, Utrecht University, 2Image Sciences Institute, University Medical Center Utrecht, 3BrainNow Research Institute, 4Institute for Stroke and Dementia Research, University Hospital, LMU Munich

Provided here is a practical tutorial for an open-access, standardized image processing pipeline for the purpose of lesion-symptom mapping. A step-by-step walkthrough is provided for each processing step, from manual infarct segmentation on CT/MRI to subsequent registration to standard space, along with practical recommendations and illustrations with exemplary cases.

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