title: Visualization of postmortem chondrocyte damage by vital staining and confocal laser scanning 3D microscopy.
abstract: The present study was designed to investigate whether the combination of vital dyes [calcein acetomethyl ester and ethidium homodimer (LIVE/DEAD Viability/Cytoxicity Kit)] together with confocal laser scanning 3D microscopy was a suitable process to detect postmortem chondrocyte damage, and whether this process could be used to establish postmortem interval. Human knee cartilage from 13 autopsies (postmortem interval from 1 day to 2.5 months) was incubated with the two dyes. The chondrocytes revealed intense staining according to their vitality. For those cases that were stored mainly at 4 degrees C there was a vitality of approximately 88 to 96% within the first 4.5 days, which decreased to 58% after 6 days and to 9% after 1.5 months. After 2 days and 14 days at summer temperatures there were 70% and 8% vital chondrocytes respectively. Three of the 13 cases showed that altered body and storage conditions limited the efficacy of the method. Initial data suggested a time and temperature dependent increase in cell breakdown. Under stable cooling conditions the use of vital dyes and confocal laser scanning 3D microscopy to measure chondrocyte loss may be a valuable tool for estimating the postmortem interval.
authors.affiliation: Institute of Legal Medicine, University of Giessen, Germany.
authors.forename: Gabriele E
authors.initials: GE
authors.lastname: Lasczkowski
authors.forename: Thomas
authors.initials: T
authors.lastname: Aigner
authors.forename: Ulrike
authors.initials: U
authors.lastname: Gamerdinger
authors.forename: Gunther
authors.initials: G
authors.lastname: Weiler
authors.forename: Hansjürgen
authors.initials: H
authors.lastname: Bratzke
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002356:Cartilage / Q000166:cytology / Q000473:pathology*; D019902:Chondrocytes / Q000473:pathology*; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D008297:Male; D018613:Microscopy, Confocal; D008875:Middle Aged; D011180:Postmortem Changes
chemical_list: D005456:Fluorescent Dyes