Early detection of apoptosis by staining of acid-treated apoptotic cells with FITC-labeled lectin from Narcissus pseudonarcissus.
BACKGROUND
Exposure of anionic phospholipids and modified carbohydrates are main parts of the apoptotic death program. Cells undergoing apoptosis can be identified by various methods, detecting surface changes or modifications of their organelles, respectively. We describe a method for the detection of early apoptosis by staining of cells with fluorescein isothiocyanate (FITC)-labeled lectin from Narcissus pseudonarcissus (NPn).


METHODS
Apoptosis in cells or in cell lines was induced by various stimuli. To detect apoptosis the cells were stained with FITC-labeled lectin of NPn. After a short-term acid treatment they were analyzed by flow cytometry.


RESULTS
The instability of the cytoplasmic membrane against acid and the binding of NPn were very early features of apoptotic cell death. The NPn lectin staining procedure detected apoptosis with high sensitivity. The staining was stable for at least 12 h.


CONCLUSIONS
The method described in this study is suitable for the detection of the very early phases of apoptosis. The NPn lectin staining after short-term acid treatment can, therefore, be added to the list of reliable tools for the research of cell death.
D000143:Acids / Q000494:pharmacology*; D017304:Annexin A5 / Q000737:chemistry; D017209:Apoptosis / Q000187:drug effects* / Q000528:radiation effects; D015415:Biomarkers / Q000737:chemistry; D045744:Cell Line, Tumor; D005434:Flow Cytometry / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D006801:Humans; D019169:Jurkat Cells; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000528:radiation effects; D031427:Narcissus / Q000737:chemistry; D037121:Plant Lectins / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*; D013997:Time Factors; D020298:U937 Cells; D014466:Ultraviolet Rays
D000143:Acids; D017304:Annexin A5; D015415:Biomarkers; D037121:Plant Lectins; D016650:Fluorescein-5-isothiocyanate