Masking background fluorescence and luminescence in the...

Chemistry: analytical and immunological testing – Optical result

Reexamination Certificate

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C436S056000, C436S063000, C436S166000, C436S151000, C436S169000, C436S170000, C436S172000, C436S517000, C436S518000, C436S524000, C436S528000, C436S531000, C436S534000, C436S535000, C436S800000, C436S805000, C436S823000, C435S029000, C435S034000, C356S317000, C356S318000, C356S417000, C250S458100, C250S459100, C250S461200

Reexamination Certificate

active

07615376

ABSTRACT:
In a process for the quantitative optical analysis of fluorescently labelled biological cells5, a cell layer on a transparent support at the bottom2of a reaction vessel1is in contact with a solution3containing the fluorescent dye4. The sensitivity of analytical detection can be considerably improved if to the fluorescent dye4already present in addition a masking dye9, which absorbs the excitation light6for the fluorescent dye4and/or its emission light7, is added to the solution3and/or if a separating layer10permeable to the solution and absorbing and/or reflecting the excitation light6or the emission light7is applied to the cell layer at the bottom2. This process can also be used for improving the sensitivity in the quantitative optical analysis of a luminescent biological cell layer. The separating layer10must in this case be composed such that it has a high power of reflection for the luminescent light11. Analogously, these process principles can also be used in receptor studies for the masking of the interfering background radiation in the quantitative optical analysis of fluorescently or luminescently labelled reaction components. In this case, a receptor layer12at the bottom2of a reaction vessel1is in contact with a solution (supernatant3) in which a fluorescent or luminescent ligand13is dissolved. The sensitivity and accuracy of the analytical detection can be considerably improved here if a masking dye9which absorbs the excitation light6for the fluorescent dye and/or its emission light or (in the case of luminescent ligands) the luminescent light is added to the supernatant3. Instead of the masking dye in the solution3or optionally as an additional measure, a separating layer10permeable to the solution3and absorbing and/or reflecting the excitation light6and/or the emission light or the luminescent light can be applied to the cell or receptor layer12at the bottom2.

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