AI Article Synopsis
- The study measures how the fluorescence yield of stained micro beads changes with different laser beam intensities, crucial for flow cytometry calibration.
- A rate equation model helps estimate the number of fluorescent molecules per micro bead, using specific dye properties obtained through a flow cytometric sorting technique.
- Initial results demonstrate the effectiveness of this method by showing fluorescence saturation and single molecule bleaching observed under high sensitivity microscopy.
Article Abstract
We have determined the fluorescence yield of stained micro beads, used for calibration purposes in flow cytometry, as function of the irradiance of the exciting laser beam. A rate equation model has been applied to derive the number of fluorescence molecules carried by each micro bead. To derive in situ photo-physical properties of the specific dye, required for the rate equation model, we discuss an approach based on flow cytometric sorting of micro beads, which have passed two laser beams with properly chosen different irradiances, and subsequent observation of single molecule bleaching employing high sensitivity microscopy. The feasibility of our approach is demonstrated presenting first results concerning saturation of fluorescence of beads in flow and single molecule bleaching by high sensitivity microscopy.
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| http://dx.doi.org/10.1007/s10895-005-2635-y | DOI Listing |
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