Rapid automated multichannel microspectrofluorometry. A new method for studies on the cell-to-cell transfer of molecules.

摘要: Abstract A new microchemical method is described to study the transfer of molecules between neighboring cells: rapid multichannel microfluorometry. The cell-to-cell rates metabolites or fluorescent tracers (microinjected by a microelectrophoretic piezoelectric technique) are followed at pace with in situ velocities such processes via multisite topographic monitoring fluorescence injected cell and neighbors (continuously before, during after microinjection). Furthermore, sensitivity allows kinetic using (e.g., glucose-6-P) which not themselves, but elicit associated changes redox state endogenous fluorochromes, i.e. NAD(P) ⇆ NAD(P)H transients. Since chemicals known proceed various cells tissues intercellular junctions, above technique was applied coupled system (e.g. Chironomus salivary gland) an uncoupled L cells). fluorescein observable kinetically glands, liver, Chang liver conjunctiva cultures. Equilibration dye neighbor completed within few hundred msec sec, corresponding half total flux per unit concentration difference ( g f ) ~2–10 × 10 4 μm 3 /min. Such practically basent rare occurrence cells. In NCTC 8739 cells, glucose-6-P seemed occur more frequently than that fluorescein, suggesting possibility (or catabolite) may move through non-junctional regions membrane. Thus microfluorometry makes possible tracer metabolite transfer, without need for multiple implantation electrodes detail obtainable other techniques.