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3 ARTICLES PUBLISHED IN JoVE

JoVE Journal

Identifying DNA Mutations in Purified Hematopoietic Stem/Progenitor Cells

Ziming Cheng¹, Ting Zhou^1,2, Azhar Merchant¹, Thomas J. Prihoda³, Brian L. Wickes^4,5, Guogang Xu², Christi A. Walter^2,5, Vivienne I. Rebel^1,2,5

¹Greehey Children's Cancer Research Institute, UT Health Science Center at San Antonio, ²Department of Cellular and Structural Biology, UT Health Science Center at San Antonio, ³Department of Pathology, UT Health Science Center at San Antonio, ⁴Department of Microbiology, UT Health Science Center at San Antonio, ⁵Cancer Therapy and Research Center, UT Health Science Center at San Antonio

Here we describe an in vivo mutagenesis assay for small numbers of purified hematopoietic cells using the LacI transgenic mouse model. The LacI gene can be isolated to determine the frequency, location, and type of DNA mutants spontaneously arisen or after exposure to genotoxins.

JoVE Core

Quality-Controlled Sputum Analysis by Flow Cytometry

Marcia Grayson^*1, Shao-Chiang Lai^*1, Lydia H. Bederka¹, Patricia Araujo¹, Jamila Sanchez¹, Xavier T. Reveles¹, Vivienne I. Rebel^1,2, Jennifer Rebeles¹

¹bioAffinity Technologies, ²Department of Cell Systems & Anatomy, The University of Texas Health Science Center at San Antonio

This protocol describes an efficient method for dissociating sputum into a single cell suspension and the subsequent characterization of cellular subsets on standard flow cytometric platforms.

Chemistry

Porphyrin-Modified Beads for Use as Compensation Controls in Flow Cytometry

William Bauta¹, Marcia Grayson¹, Rossella Titone¹, Jennifer Rebeles¹, Vivienne I. Rebel¹

¹bioAffinity Technologies

The protocol describes how porphyrin-based compensation beads for flow cytometry are prepared by the reaction of amine-functionalized polystyrene beads with the porphyrin TCPP and the amide coupling reagent EDC. A filtration procedure is used to reduce the particulate byproducts.