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Doube, Michael on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Tetracycline-to-bone-augmentation-tp7580337p7580340.html
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Dear Javier,
> We want to evaluate the bone augmentation in rabbit tibia using
> Tetracycline or similar.
I think you are referring to the very old technique of in-vivo labelling
of mineral apposition in calcified tissues, right? In which case you
might consider other labels such as alizarin and calcein (green and
blue). Calcein green in particular gives a stable bright green
fluorescence and works well with fluorescein filters and excitation with
a 488nm laser. Main issue is the yellowish autofluorescence from the
bone matrix and cells themselves. But, that, and the backscattered
reflected light can be used to your advantage to give tissue contrast
without further processing. See e.g. (shameless self-promotion warning):
Doube, M., Firth, E. C. & Boyde, A. Variations in articular calcified
cartilage by site and exercise in the 18-month-old equine distal
metacarpal condyle. Osteoarth Cart 15, 1283–1292 (2007).
http://dx.doi.org/10.1016/j.joca.2007.04.003And the originals:
1.FROST, H. M. Measurement of human bone formation by means of
tetracycline labelling. Can J Biochem Physiol 41, 31–42 (1963).
2.FROST, H. M. Tetracycline labelling of bone and the zone of
demarcation of osteoid seams. Can J Biochem Physiol 40, 485–489 (1962).
There is lots more literature on labelling protocols, see e.g.
Erben, R. G. & Glösmann, M. Histomorphometry in rodents. Methods Mol.
Biol. 816, 279–303 (2012).
http://dx.doi.org/10.1007/978-1-61779-415-5_19Michael
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