Characterization of eight different tetracyclines: advances in fluorescence bone labeling
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Characterization of eight different tetracyclines: advances in fluorescence bone labeling. / Pautke, Christoph; Vogt, Stephan; Kreutzer, Kilian; Haczek, Cornelia; Wexel, Gabriele; Kolk, Andreas; Imhoff, Andreas B; Zitzelsberger, Horst; Milz, Stefan; Tischer, Thomas.
In: J ANAT, Vol. 217, No. 1, 07.2010, p. 76-82.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Characterization of eight different tetracyclines: advances in fluorescence bone labeling
AU - Pautke, Christoph
AU - Vogt, Stephan
AU - Kreutzer, Kilian
AU - Haczek, Cornelia
AU - Wexel, Gabriele
AU - Kolk, Andreas
AU - Imhoff, Andreas B
AU - Zitzelsberger, Horst
AU - Milz, Stefan
AU - Tischer, Thomas
PY - 2010/7
Y1 - 2010/7
N2 - 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.
AB - 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.
KW - Animals
KW - Bone and Bones
KW - Feasibility Studies
KW - Fluorescent Dyes
KW - Image Processing, Computer-Assisted
KW - Male
KW - Microscopy, Confocal
KW - Microscopy, Fluorescence
KW - Rats
KW - Rats, Wistar
KW - Tetracyclines
U2 - 10.1111/j.1469-7580.2010.01237.x
DO - 10.1111/j.1469-7580.2010.01237.x
M3 - SCORING: Journal article
C2 - 20456523
VL - 217
SP - 76
EP - 82
JO - J ANAT
JF - J ANAT
SN - 0021-8782
IS - 1
ER -