title: Development of a microbial high-throughput screening instrument based on elastic light scatter patterns.
abstract: A microbial high-throughput screening (HTS) system was developed that enabled high-speed combinatorial studies directly on bacterial colonies. The system consists of a forward scatterometer for elastic light scatter (ELS) detection, a plate transporter for sample handling, and a robotic incubator for automatic incubation. To minimize the ELS pattern-capturing time, a new calibration plate and correction algorithms were both designed, which dramatically reduced correction steps during acquisition of the circularly symmetric ELS patterns. Integration of three different control software programs was implemented, and the performance of the system was demonstrated with single-species detection for library generation and with time-resolved measurement for understanding ELS colony growth correlation, using Escherichia coli and Listeria. An in-house colony-tracking module enabled researchers to easily understand the time-dependent variation of the ELS from identical colony, which enabled further analysis in other biochemical experiments. The microbial HTS system provided an average scan time of 4.9 s per colony and the capability of automatically collecting more than 4000 ELS patterns within a 7-h time span.
authors.affiliation: School of Mechanical Engineering, Purdue University, West Lafayette, Indiana 47907, USA.
authors.forename: Euiwon
authors.initials: E
authors.lastname: Bae
authors.forename: Valery
authors.initials: V
authors.lastname: Patsekin
authors.forename: Bartek
authors.initials: B
authors.lastname: Rajwa
authors.forename: Arun K
authors.initials: AK
authors.lastname: Bhunia
authors.forename: Cheryl
authors.initials: C
authors.lastname: Holdman
authors.forename: V Jo
authors.initials: VJ
authors.lastname: Davisson
authors.forename: E Daniel
authors.initials: ED
authors.lastname: Hirleman
authors.forename: J Paul
authors.initials: JP
authors.lastname: Robinson
mesh_terms: D002138:Calibration; D046508:Culture Techniques; D004548:Elasticity; D004926:Escherichia coli / Q000254:growth & development / Q000302:isolation & purification*; D008027:Light; D008087:Listeria / Q000254:growth & development / Q000302:isolation & purification*; D012542:Scattering, Radiation; D012984:Software; D013997:Time Factors