We have studied physiological parameters in a living cell using fluorescence lifetime imaging of endogenous chromophores. In this study, pH dependence of the fluorescence lifetime of flavin adenine dinucleotide (FAD), that is a significant cofactor exhibiting autofluorescence, has been investigated in buffer solution and in cells. The fluorescence lifetime of FAD remained unchanged with pH 5 to 9 in solution. However, the fluorescence lifetime in HeLa cells was found to decrease with increasing intracellular pH, suggesting that pH in a single cell can be estimated from the fluorescence lifetime imaging of FAD without adding exogenous fluorescent probes.
Own experiences with the identification of mice and rats using ear tags, ear punching, toe clipping (for newborns only) and ear tattooing are given and compared with methods presented in literature.By far, ear tattooing by the newly developed pincers is the most suitable method for marking adult rodents for life. Tattooing is easy to apply, easy to read and not harmful to the animals. For newborn pups on day 4 p. p., the s, c, injection of India ink into the palm of paw is recommended.
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