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Methodological aspects of flow cytometric analysis of DNA polyploidy in human heart tissue

โœ Scribed by H. W. Vliegen; A. M. Vossepoel; A. Laarse; F. Eulderink; C. J. Cornelisse


Book ID
104783545
Publisher
Springer
Year
1986
Tongue
English
Weight
643 KB
Volume
84
Category
Article
ISSN
1432-119X

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โœฆ Synopsis


The purpose of the study was to investigate the possibilities of flow cytometry (FCM) for the analysis of DNA polyploidy in human heart tissue. Suspensions of single nuclei were prepared with the detergenttrypsin procedure and stained with propidium iodide. A mathematical correction procedure was developed to correct for background and clumping. For diploid model populations of chicken and trout red blood cells this correction reduced artifactual fractions in the FCM DNA profile to less than 0.5 %, indicating that interference by background and clumping was almost completely overcome by this correction procedure. FCM DNA profiles were obtained from 12 hypertrophic and 7 normal human adult hearts. Clear differences were found between the DNA profiles from the normal and the hypertrophic hearts, the latter showing a higher degree of polyploidization. From the corrected DNA profiles, six different polyploidization parameters were computed, all of which showed a significant correlation with at least three out of four different parameters for heart hypertrophy. FCM appears to be a reliable method for the measurement of polyploidization in heart tissue, provided background and clumping are corrected for.


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