Journal article
Visualizing lens epithelial cell proliferation in whole lenses
Molecular vision, Vol.16(138-39), pp.1253-1259
07/09/2010
PMCID: PMC2903465
PMID: 20664699
Abstract
To develop a means to image cells in S-phase of the cell cycle while preserving the anatomic relationships within the lens. Mice were injected with the thymidine analog, EdU. Whole lenses were removed, fixed and permeabilized. Cells that had incorporated EdU into their DNA were chemically labeled using fluorescent azides and "click" chemistry. Double labeling was performed with antibodies to other antigens, like phospho-histoneH3, a marker of mitotic cells. The position of labeled cells and lens anatomy was viewed using a simple device to position and flatten the lens. The nuclei of cells in S-phase of the cell cycle were intensely stained without the use of antibodies. Stained cells were readily localized with reference anatomic landmarks, like the transition zone. Whole lenses could be assayed by rotating the lens on the microscope stage. Double-labeling permitted the co-localization of markers in cycling cells. EdU labeling of whole lenses provides a simple, rapid and sensitive means to analyze lens epithelial cell proliferation in the anatomic context of the whole lens.
Details
- Title: Subtitle
- Visualizing lens epithelial cell proliferation in whole lenses
- Creators
- Luke A Wiley - Department of Ophthalmology and Visual Sciences, Washington University, Saint Louis, MO 63110, USA. wileyl@vision.wustl.eduYing-Bo ShuiDavid C Beebe
- Resource Type
- Journal article
- Publication Details
- Molecular vision, Vol.16(138-39), pp.1253-1259
- Publisher
- United States
- PMID
- 20664699
- PMCID
- PMC2903465
- ISSN
- 1090-0535
- eISSN
- 1090-0535
- Grant note
- T32 HL007873 / NHLBI NIH HHS P30 EY002687 / NEI NIH HHS R01 EY004853 / NEI NIH HHS EY04853 / NEI NIH HHS EY002687 / NEI NIH HHS
- Language
- English
- Date published
- 07/09/2010
- Academic Unit
- Ophthalmology and Visual Sciences
- Record Identifier
- 9983980386102771
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