Fluorescence instability of propidium iodide-labelled microspheres in (para)formaldehyde
| Autor: | Walter Kaffenberger, Edgar L. Rohr |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Polymers
Biophysics Formaldehyde Conjugated system Fluorescence Pathology and Forensic Medicine Flow cytometry chemistry.chemical_compound Endocrinology Leukocytes medicine Humans Sample preparation Propidium iodide Paraformaldehyde Chromatography medicine.diagnostic_test Cell Biology Hematology Flow Cytometry Microspheres Phenotype chemistry Quantitative analysis (chemistry) Propidium |
| Zdroj: | Cytometry. 14:584-586 |
| ISSN: | 1097-0320 0196-4763 |
| DOI: | 10.1002/cyto.990140520 |
| Popis: | An instability of the fluorescence of propidium iodide (PI)-labelled microspheres was observed when the beads were used as an internal fluorescence standard in cell samples suspended in paraformaldehyde (pFA) for flow cytometric phenotyping. Flow cytometry of fluorochrome-labelled microspheres as well as spectrofluorometry of PI-fluorochrome solutions in PBS-buffered 1% pFA or formalin (FA) revealed strong increases of PI fluorescence starting immediately after the addition to the fixatives. We propose a chemical reaction which leads to two additional conjugated double bonds in the modified fluorochrome causing the increased fluorescence emission. Therefore, PI-labelled microspheres should not be applied as internal fluorescence references in aldehyde-containing cell suspensions for flow cytometric analysis, except when the time interval between the addition of beads and the measurement can be kept constant or if it lasts longer than 1 h. |
| Databáze: | OpenAIRE |
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