Access

You are not currently logged in.

Access your personal account or get JSTOR access through your library or other institution:

login

Log in to your personal account or through your institution.

If You Use a Screen Reader

This content is available through Read Online (Free) program, which relies on page scans. Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.

Characterization and Efficacy of PKH26 as a Probe to Study the Replication History of the Human Hematopoietic KG1a Progenitor Cell Line

Gyun Min Lee, Stephen S. Fong, Duk Jae Oh, Karl Francis and Bernhard O. Palsson
In Vitro Cellular & Developmental Biology. Animal
Vol. 38, No. 2 (Feb., 2002), pp. 90-96
Stable URL: http://www.jstor.org/stable/4295312
Page Count: 7
  • Read Online (Free)
  • Subscribe ($19.50)
  • Cite this Item
Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.
Characterization and Efficacy of PKH26 as a Probe to Study the Replication History of the Human Hematopoietic KG1a Progenitor Cell Line
Preview not available

Abstract

The PKH26 dye can, in principle, be used for the study of asymmetric cell divisions (ASDs). A requirement for the identification of ASDs based on fluorescence intensity is that the PKH26 dye is distributed equally between daughter cells at each division, but this has not been demonstrated at a single-cell level. The efficacy of PKH26 as a probe for the study of ASDs was examined using the human hematopoietic KG1a cell. An automated time-lapse fluorescent microscope system was used to determine changes in cell size and fluorescence intensity during culture, and track cell divisions. The images of daughter cells were analyzed using the Isee™ software to determine the distribution of PKH26 dye between daughter cells. Ratios of cell size, mean fluorescence intensity, and total fluorescence intensity were calculated by dividing the values for one daughter cell by the value of the other daughter cell. The ratios for cell size, mean intensity, and total intensity were 1.13 ± 0.12, 1.08 ± 0.07, and 1.15 ± 0.14 (mean ± SD), respectively. Thus, PKH26 is not distributed equally to both daughter cells upon cell division. However, the replication history of individual KG1a cells can be reliably deduced for up to three divisions based solely on the mean and total fluorescence intensity of the PKH26 dye, using PKH26 concentrations below the chemical and phototoxic limits (2 µM).

Page Thumbnails

  • Thumbnail: Page 
90
    90
  • Thumbnail: Page 
91
    91
  • Thumbnail: Page 
92
    92
  • Thumbnail: Page 
93
    93
  • Thumbnail: Page 
94
    94
  • Thumbnail: Page 
95
    95
  • Thumbnail: Page 
96
    96