title: High speed classification of individual bacterial cells using a model-based light scatter system and multivariate statistics.
abstract: We describe a model-based instrument design combined with a statistical classification approach for the development and realization of high speed cell classification systems based on light scatter. In our work, angular light scatter from cells of four bacterial species of interest, Bacillus subtilis, Escherichia coli, Listeria innocua, and Enterococcus faecalis, was modeled using the discrete dipole approximation. We then optimized a scattering detector array design subject to some hardware constraints, configured the instrument, and gathered experimental data from the relevant bacterial cells. Using these models and experiments, it is shown that optimization using a nominal bacteria model (i.e., using a representative size and refractive index) is insufficient for classification of most bacteria in realistic applications. Hence the computational predictions were constituted in the form of scattering-data-vector distributions that accounted for expected variability in the physical properties between individual bacteria within the four species. After the detectors were optimized using the numerical results, they were used to measure scatter from both the known control samples and unknown bacterial cells. A multivariate statistical method based on a support vector machine (SVM) was used to classify the bacteria species based on light scatter signatures. In our final instrument, we realized correct classification of B. subtilis in the presence of E. coli,L. innocua, and E. faecalis using SVM at 99.1%, 99.6%, and 98.5%, respectively, in the optimal detector array configuration. For comparison, the corresponding values for another set of angles were only 69.9%, 71.7%, and 70.2% using SVM, and more importantly, this improved performance is consistent with classification predictions.
authors.affiliation: School of Mechanical Engineering, Purdue University, West Lafayette, IN 47907, USA. mvenkata@purdue.edu
authors.forename: Murugesan
authors.initials: M
authors.lastname: Venkatapathi
authors.forename: Bartek
authors.initials: B
authors.lastname: Rajwa
authors.forename: Kathy
authors.initials: K
authors.lastname: Ragheb
authors.forename: Padmapriya P
authors.initials: PP
authors.lastname: Banada
authors.forename: Todd
authors.initials: T
authors.lastname: Lary
authors.forename: J Paul
authors.initials: JP
authors.lastname: Robinson
authors.forename: E Daniel
authors.initials: ED
authors.lastname: Hirleman
mesh_terms: D001185:Artificial Intelligence; D001412:Bacillus subtilis / Q000145:classification* / Q000166:cytology / Q000302:isolation & purification; D015373:Bacterial Typing Techniques / Q000379:methods*; D001699:Biometry; D013293:Enterococcus faecalis / Q000145:classification* / Q000166:cytology / Q000302:isolation & purification; D004867:Equipment Design; D004926:Escherichia coli / Q000145:classification* / Q000166:cytology / Q000302:isolation & purification; D005434:Flow Cytometry; D008027:Light; D008087:Listeria / Q000145:classification* / Q000166:cytology / Q000302:isolation & purification; D008954:Models, Biological; D015999:Multivariate Analysis; D010363:Pattern Recognition, Automated / Q000379:methods; D012031:Refractometry; D012542:Scattering, Radiation