Abstract
Fluorochrome-labelled bacteria were tested for their rosette-forming properties with human lymphocytes in suspension. Acridine orange stained human buffy coat cells or isolated mononuclear cells are rosetted with tetramethyl-rhodamine-isothiocyanate (TRITC)-labelled bacterial strains alone (mono-bacterial rosetting test) or simultaneously with a TRITC-labelled strain and a mutant or taxonomically different strain, labelled with fluorescein-isothiocyanate (double bacterial rosetting test [D-BRT]). Suspensions are centrifuged, washed, finally counterstained with ethidium bromide and examined by fluorescence microscopy. The bacterial binding properties of B cells may be studied by using anti-Ig pretreated mononuclear cells and TRITC bacteria (anti-Ig/BRT). In this study the methodology for bacterial staining, mono-, double- and anti-Ig/BRT is given. Estimation of rosette-forming cells is very accurate, easy and quick due to the bright fluorescence of the bacterial 'beads'. Furthermore, broad applicability of the bacterial rosetting phenomenon to study lymphocyte heterogeneity is gained with the fluorescent assay system.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
