Simple adaptations to extend the range of flow cytometry five orders of magnitude for the DNA analysis of uni- and multicellular systems
Procedures and instrumentation are described to extend the capability of a cytometry system to record samples that exhibit a wide range of fluorescence such as multicellular systems. The method employs a log amplifier in combination with a set of neutral density filters that reduces the incident light reaching the photomultiplier tube. With any given filter, signals within an intensity range of 200-fold can be measured; different filters can be used to obtain an extended overall range. Polystyrene fluorescent microspheres and a variety of mithramycin stained biological samples ranging from yeast cells to Paramecium were processed by the system. The relative DNA content of individual multicellular embryos was determined for a heterogeneous population of embryonic stages isolated from the nematoda, Caenorhahditis elegans. As part of the evaluation of the procedure, the practical upper limit of range extension was determined. The most intense fluorescent signal was produced when untreated pecan pollen stained with ethidium bromide fluoresced with a factor (8.4 +- 1.3) x 10/sup 4/ more than ethidium bromide stained E. coli cells.
- Research Organization:
- Univ. of Houston, TX
- OSTI ID:
- 5987346
- Journal Information:
- J. Histochem. Cytochem.; (United States), Vol. 29:6
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
37 INORGANIC
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
CELL FLOW SYSTEMS
CALIBRATION
DNA
QUANTITATIVE CHEMICAL ANALYSIS
ANIMAL CELLS
COMPARATIVE EVALUATIONS
CYTOLOGICAL TECHNIQUES
DENSITY
EMBRYOS
MEASURING METHODS
MECHANICAL FILTERS
POLLEN
SAMPLING
CHEMICAL ANALYSIS
FILTERS
GAMETES
GERM CELLS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PHYSICAL PROPERTIES
550300* - Cytology
400102 - Chemical & Spectral Procedures