Measurement of intracellular concentration of fluorescently-labeled targets in living cells

Estimations of intracellular concentrations of fluorescently-labeled molecules within living cells are very important for guidance of biological experiments and interpretation of their results. Here we propose a simple and universal approach for such estimations. The approach is based upon common knowledge that the dye fluorescence is directly proportional to its quantum yield and the number of its molecules and that a coefficient of proportionality is determined by spectral properties of the dye and optical equipment used to record fluorescent signals. If two fluorescent dyes are present in the same volume, then a ratio of their concentrations is equal to a ratio of their fluorescence multiplied by some dye- and equipment-dependent coefficient. Thus, if the coefficient and concentration of one dye is known then the concentration of another dye can be determined. Here we have demonstrated how to calculate this coefficient (called a ratio factor) and how to use it for concentration measurements of fluorescently tagged molecules. As an example of how this approach can be used, we estimated a concentration of exogenously expressed neuronal Ca2+ sensor protein, hippocalcin, tagged by a fluorescent protein in a dendritic tree of rat hippocampal neurons loaded via a patch pipette with Alexa Fluor dye of known concentration. The new approach should allow performing a fast, inexpensive and reliable quantitative analysis of fluorescently-labeled targets in different parts of living cells

The super fixed target beauty facility at the SSC

The rationale for pursuing beauty physics at the SSC in a fixed target configuration is described. The increased beauty production cross section at the SSC, combined with high interaction rate capability of the proposed detector, results in 1010-11 produced BB events per year. The long decay length of the B hadrons (all equal to10 cm) allows direct observation of B decays in the high resolution silicon microstrip vertex detector. To optimize the operation of the proposed beauty spectrometer and the SSC, parasitic extraction of attendant or artificially generated large amplitude protons using crystal channeling is proposed and explored. The large sample of fully reconstructed B events allows detailed studies various CP violating decays with requisite statistics to confront the standard model. The CP physics potentials of the proposed experiment is evaluated and compared with alternative approaches, such as asymmetric e+e- B Factories and specialized hadron colliders. © 1992