Instrumentation and Analyses for Multi-Angle Light Scatter Measurements in Flow Cytometry

MULTIPARAMETER FLOW CYTOMETRIC INSTRUMENTATION HAS BEEN DESIGNED AND CONSTRUCTED IN COLLABORATION BETWEEN JRC-ISPRA AND THE DEPARTMENT OF PHYSICS AND APPLIED PHYSICS, UNIVERSITY OF STRATHCLYDE TO ALLOW THE EXTENSION OF SINGLE CELL DIFFRACTOMETRY TO FLOW SYSTEMS. THE SYSTEM MEASURES UP TO 16 PARAMETERS SIMULTANEOUSLY, OF WHICH 2 MAY BE FLUORESCENCE EMISSION AND THE REMAINDER LIGHT SCATTER IN UP TO 16 ANGULAR WINDOWS IN THE FORWARD AND 90 DEGREES DIRECTIONS. THE INSTRUMENTATION UTILISES PHOTODIODE ARRAYS WHICH ARE SO MOUNTED AS TO ALLOW USER SELECTION OF THE ANGULAR ACCEPTANCE WINDOWS OF THE DETECTORS. THIS FACILITATES THE DISTIN- CTION OF ALGAL GENERA ON THE BASIS OF MORPHOLOGICAL CHARACTERISTICS, THUS WIDENING THE FIELD OF POSSIBLE ECONOLOGICAL APPLICATIONS OF FLOW CYTOMETRY. LABORATORY EXPERIMENTS HAVE BEEN PERFORMED IN ORDER TO TEST THE DESCRIMINA- TORY ABILITY OF THE TECHNIQUE USING BOTH MARINE AND FRESHWATER ALGAE AND ALSO TO ASSESS ITS VALUE FOR APPLICATION TO MULTI-SPECIES BIOASSAYS. IN ADDITION, METHODS OF DATA ANALYSIS ARE UNDER DEVELOPMENT WHICH SHOULD BE APPLICABLE IN OTHER FIELDS OF MULTIPARAMETER FLOW CYTOMETRY. TO DATE DISCRIMINANT ANALYSIS TECHNIQUES HAVE BEEN DEVELOPED WHICH ALLOW DISTIN- CTION AMONG CERTAIN CLASSES OF ALGAE, BUT FURTHER DEVELOPMENT IS NECESSARY.

PREMAZZI Guido;  BUONACCORSI Giovanni;  CUNNINGHAM A.;  FORREST J.;  MAAS Peter; 

1995-03-15

JRC5023

https://publications.jrc.ec.europa.eu/repository/handle/JRC5023,