title: Characterization of eight different tetracyclines: advances in fluorescence bone labeling.
abstract: Polychrome sequential labeling with fluorochromes is a standard technique for the investigation of bone formation and regeneration processes in vivo. However, for human application, only tetracycline and its derivates are approved as fluorochromes. Therefore, the aim of this study was to determine the fluorescence characteristics of the different tetracycline derivates to assess the feasibility of sequential in vivo bone labeling using distinguishable fluorochromes. Eight different tetracycline derivates were injected subcutaneously into growing rats as a single dose or sequentially in different combinations. After preparation of resin-embedded undecalcified bone sections, the fluorescence properties of the tetracycline derivates in bone were analyzed using conventional fluorescence microscopy, spectral image analysis and confocal laser scanning microscopy. Each tetracycline derivate exhibited a characteristic fluorescence spectrum, but the differences between them were small. Chlortetracycline could be discriminated reliably from all other derivates and could therefore be combined with any other tetracycline derivate for reliably distinguishable double labeling. Tetracycline itself exhibited the brightest fluorescence of all the investigated derivates. Interestingly, in conventional microscopy the same tetracycline derivative can appear in different colours to the human eye, even if spectral analysis confirmed identical emission peaks. In conclusion, the data suggest that fluorescence double labeling of bone is feasible using appropriate tetracycline derivates in combination with spectral imaging modalities.
authors.affiliation: Department of Oral and Maxillofacial Surgery, University of Munich, Munich, Germany. christoph.pautke@gmx.net
authors.forename: Christoph
authors.initials: C
authors.lastname: Pautke
authors.forename: Stephan
authors.initials: S
authors.lastname: Vogt
authors.forename: Kilian
authors.initials: K
authors.lastname: Kreutzer
authors.forename: Cornelia
authors.initials: C
authors.lastname: Haczek
authors.forename: Gabriele
authors.initials: G
authors.lastname: Wexel
authors.forename: Andreas
authors.initials: A
authors.lastname: Kolk
authors.forename: Andreas B
authors.initials: AB
authors.lastname: Imhoff
authors.forename: Horst
authors.initials: H
authors.lastname: Zitzelsberger
authors.forename: Stefan
authors.initials: S
authors.lastname: Milz
authors.forename: Thomas
authors.initials: T
authors.lastname: Tischer
mesh_terms: D000818:Animals; D001842:Bone and Bones / Q000378:metabolism*; D005240:Feasibility Studies; D005456:Fluorescent Dyes; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008297:Male; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D051381:Rats; D017208:Rats, Wistar; D013754:Tetracyclines
chemical_list: D005456:Fluorescent Dyes; D013754:Tetracyclines
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