Titre du document / Document title

Polychrome labeling of bone with seven different fluorochromes : Enhancing fluorochrome discrimination by spectral image analysis

Auteur(s) / Author(s)

PAUTKE Christoph ⁽¹⁾ ; VOGT Stephan ⁽²⁾ ; TISCHER Thomas ⁽²⁾ ; WEXEL Gabriele ⁽³⁾ ; DEPPE Herbert ⁽¹⁾ ; MILZ Stefan ⁽⁴⁾ ; SCHIEKER Matthias ⁽⁵⁾ ; KOLK Andreas ⁽¹⁾ ;

Affiliation(s) du ou des auteurs / Author(s) Affiliation(s)

⁽¹⁾ Department of Oral and Cranio-Maxillofacial Surgery, Klinikum rechts der Isar, Technical University of Munich (TUM), Ismaninger Str. 22, 81675 Munich, ALLEMAGNE
⁽²⁾ Department of Orthopedic Sports Medicine, TUM, Ismaninger Str. 22, 81675 Munich, ALLEMAGNE
⁽³⁾ Institut für Experimentelle Onkologie und Therapieforschung, TUM, Ismaninger Str. 22, 81675 Munich, ALLEMAGNE
⁽⁴⁾ AO Research Institute, Clavadelerstrasse, 7270 Davos, SUISSE
⁽⁵⁾ Department of Surgery, Ludwig-Maximilians-University (LMU) Munich, Nussbaumstr. 20, 80336 Munich, ALLEMAGNE

Résumé / Abstract

Bone formation and remodeling in vivo can be assessed by polychrome labeling using calcium-binding fluorescent dyes. The number of fluorochromes, however, limits this technique due to the fact that with increasing number, fluorescent spectra inevitably overlap, which makes discrimination more difficult. In order to enhance discrimination, we performed spectral image analysis. Non-critical size defects of the femur of male Wistar rats served as a model for bone formation. Eight different fluorochromes (calcein blue, xylenol orange, calcein, alizarine complexone, doxycycline, rolitetracycline, hematoporphyrin, and BAPTA) were administered sequentially subcutaneously every third day starting at day 4 after surgery. Following, bone specimen were embedded in methylmethacrylate and analyzed by spectral image acquisition using a Sagnac type interferometer (ASI, Israel). Seven of the eight applied fluorochromes could be resolved using spectral image examination. With BAPTA, we present a new fluorochrome suitable for bone labeling. Due to the superior sensitivity of the spectral image acquisition, the thickness of the bone sections could be reduced so that 5 μm thick sections could be analyzed. Spectral decomposition and subsequent linear unmixing allows depiction of each individual fluorochrome without interference of any other, enabling a reliable and superior morphometric analysis of labeled regions.

Revue / Journal Title

Bone   ISSN 8756-3282 

Source / Source

2005, vol. 37, n^o4, pp. 441-445 [5 page(s) (article)] (17 ref.)

Langue / Language

Anglais

Editeur / Publisher

Elsevier, New York, NY, ETATS-UNIS  (1985) (Revue)

Mots-clés anglais / English Keywords

Tetracycline derivatives ; Inorganic element ; Asia ; Treatment ; Antibiotic ; Antibacterial agent ; Israel ; Surgery ; Hematoporphyrin ; Femur ; Image analysis ; Spectral analysis ; Bone ; Rolitetracycline ; Doxycycline ; Calcium ;

Mots-clés français / French Keywords

Tétracycline dérivé ; Elément minéral ; Asie ; Traitement ; Antibiotique ; Antibactérien ; Israël ; Chirurgie ; Hématoporphyrine ; Fémur ; Analyse image ; Analyse spectrale ; Os ; Rolitétracycline ; Doxycycline ; Calcium ;

Mots-clés espagnols / Spanish Keywords

Tetraciclina derivado ; Elemento inorgánico ; Asia ; Tratamiento ; Antibiótico ; Antibacteriano ; Israel ; Cirugía ; Hematoporfirina ; Fémur ; Análisis imagen ; Análisis espectral ; Hueso ; Rolitetraciclina ; Doxiciclina ; Calcio ;

Mots-clés d'auteur / Author Keywords

Polychrome labeling ; Fluorochromes ; BAPTA ;

Localisation / Location

INIST-CNRS, Cote INIST : 19041, 35400013165411.0020