I would add that, as a general (non-specific) stain, TB works much better at
alkaline conditions (8.0 or even higher). Altough buffers can be used, we
generally make it in plain water, adjust pH, and it remains stable for a long
time (don't forget to seriously RINSE the pH electrode after adjustment;
people don't find it funny to get their solutions blue after you've been
there...). Regarding concentrations we have found very useful to have 3
concentrations around (0.01%, 0.1%, and 1.0%) to cope with the different
thickness and affinity of the material we're working with. While 1.0% is more
usefull to plastic embeded semi-thin sections (0.2-10 um) the two others cover
the range from 1 tp 30 um. Reaction hapens alomst imedaitely but having an hot
plate around helps reproducibility.
Hope it helps
Jose
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o o
o Professor Jose' A. Feijo' o
o o
o Dep. Biologia Vegetal, Fac.Ciencias Univ. Lisboa o
o Campo Grande, Ed.C2, P-1700 LISBOA, PORTUGAL o
o o
o tel. +351.1.7500069, fax +351.1.7500048 o
o e.mail: jose.feijo at fc.ul.pt o
o URL: www.fc.ul.pt/departs/biologia_vegetal/ejf.html o
o o
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