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Analysis of plating technique for viability and drug‐sensitivity determinations using Rosa cells
Author(s) -
Strauss André,
King Patrick J.
Publication year - 1981
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1981.tb00889.x
Subject(s) - plating (geology) , plating efficiency , chromatography , materials science , agar , phase (matter) , suspension (topology) , suspension culture , layer (electronics) , homogeneous , chemistry , cell culture , in vitro , composite material , biology , biochemistry , mathematics , bacteria , paleontology , genetics , organic chemistry , combinatorics , homotopy , pure mathematics
Rosa Paul's Scarlet’cell suspension cultures were used as a test system for working out a method of viability and drug‐sensitivity determination based on plating efficiency. High plating efficiencies (80–95%) were obtained on a simple synthetic medium when aggregates of a mean size of c . 100 cells/unit from exponential phase cultures were plated at a density of 1500 units/plate in the middle layer (5 ml) of three layers of the agar‐solidified medium (total = 30 ml). This 3‐layer plating technique produces homogeneous colony growth and simplifies the microscopical evaluation of plating efficiencies. The reduction of plating efficiencies seen when the smaller aggregates of stationary phase cultures were plated was mainly due to low cell density and could be overcome by enriching the medium with various supplements. Reconstitution experiments using mixtures of inactivated and non‐inactivated aggregates demonstrated that plating efficiency can be taken as a goodmeasure of viability. The described plating technique was found to be more sensitive and reliable compared to two other methods for determining p ‐fluorophenylalanine‐sensitivity of Rosa cells.