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Elastic light scatter

Elastic Light Scatter for Bacterial Recognition

Description of the technology

The majority of tools for microbial recognition and classification (e.g., ELISA, PCR) are based on physiological or genetic properties of microorganisms. However, there is an enormous interest in devising label-free and reagentless methods that would operate utilizing the biophysical signatures of microbial samples without the need for labeling and reporting biochemistry. Elastic light scattering (ELS) - one of the most fundamental optical processes whereby electromagnetic waves are forced to deviate from a straight trajectory by non-uniformities in the medium that they traverse - can be employed to provide an inexpensive, yet remarkably accurate implementation of the label-free phenotypic classification concept. The Purdue's ELS-based system provides noninvasive, label-free identification and classification of bacterial colonies in real-time. The speed of operation surpasses those offered by any existing phenotypic or metabolic classification platform.

In a semi-automated ELS reader, Petri-dishes containing bacterial colonies are placed in the plate holder, and a colony map of the plate is immediately acquired. Subsequently, a laser beam sequentially interrogates each preselected colony and generates unique scatter signature at a speed of about one colony per second (U.S. patent US7465560B2).

The acquired patterns are decomposed using unique proprietary feature extraction methodology (U.S. patent US8787633B2) and compared to the models in the library for identification utilizing machine learning techniques. Multiple studies demonstrated the ability to detect and classify pathogens from inoculated or naturally contaminated food samples validating the ELS application to biosecurity and biosurveillance.

The research use and testing of the ELS instruments at USDA-ARS laboratories resulted in generating ELS signature libraries for serovars of Salmonella and strains of Shiga-toxin producing E. coli, as well as Bacillus, Staphylococcus, Klebsiella spp., Listeria spp., and many other organisms.

Demonstration of the technology

Video presentations of various ELS platform prototypes in action

General information on ELS platform

ELS automation - https://www.youtube.com/watch?v=hE9ropMZH-E

Prototype built by Hettich GmbH - https://www.youtube.com/watch?v=7zONHoRCnFI

Label free microbial detection

Selected peer-reviewed scientific reports

Ahmed, W., Bayraktar, B., Bhunia, A., Hirleman, D., Robinson, J., Rajwa, B., 2012. Classification of bacterial contamination using image processing and distributed computing. IEEE J Biomed Health Inform 17, 232–239. https://doi.org/10.1109/TITB.2012.2222654

Akova, F., Dundar, M., Davisson, V.J., Hirleman, E.D., Bhunia, A.K., Robinson, J.P., Rajwa, B., 2010. A machine-learning approach to detecting unknown bacterial serovars. Statistical Analysis and Data Mining 3, 289–301. https://doi.org/10.1002/sam.10085

Bae, E., Kim, H., Rajwa, B., Thomas, J.G., Robinson, J.P., 2016. Current status and future prospects of using advanced computer-based methods to study bacterial colony morphology. Expert Rev. 14, 207-218. Anti-infect Ther. https://doi.org/10.1586/14787210.2016.1122524

Bae, E., Aroonnual, A., Bhunia, A.K., Hirleman, E.D., 2010a. On the sensitivity of forward scattering patterns from bacterial colonies to media composition. J. Biophoton. https://doi.org/10.1002/jbio.201000051

Bae, E., Aroonnual, A., Bhunia, A.K., Robinson, J.P., Hirleman, E.D., 2009. System automation for a bacterial colony detection and identification instrument via forward scattering. Meas. Sci. Technol. 20, 015802. https://doi.org/10.1088/0957-0233/20/1/015802

Bae, E., Bai, N., Aroonnual, A., Robinson, J.P., Bhunia, A.K., Hirleman, E.D., 2010b. Modeling light propagation through bacterial colonies and its correlation with forward scattering patterns. J. Biomed. Opt. 15, 045001. https://doi.org/10.1117/1.3463003

Bae, E., Banada, P.P., Huff, K., Bhunia, A.K., Robinson, J.P., Hirleman, E.D., 2007. Biophysical modeling of forward scattering from bacterial colonies using scalar diffraction theory. Appl Opt 46, 3639–3648. https://doi.org/10.1364/AO.46.003639

Bae, E., Banada, P.P., Huff, K., Bhunia, A.K., Robinson, J.P., Hirleman, E.D., 2006. Analysis of time-resolved scattering from macroscale bacterial colonies. J Biomed Opt 13, 014010. https://doi.org/10.1117/1.2830655

Bae, E., Ying D., Kramer, D., Patsekin, V., Rajwa, B., Holdman, C., Sturgis, J., Davison, V. J., Robinson, J.P., 2012. Portable bacterial identification system based on elastic light scatter patterns. J Bio. Eng. 6, 014010. https://doi.org/10.1186/1754-1611-6-12

Bae, E., Patsekin, V., Rajwa, B., Bhunia, A.K., Holdman, C., Davisson, V.J., Hirleman, E.D., Robinson, J.P., 2012. Development of a microbial high-throughput screening instrument based on elastic light scatter patterns. Rev Sci Instrum 83, 044304. https://doi.org/10.1063/1.3697853

Banada, P.P., Guo, S., Bayraktar, B., Bae, E., Rajwa, B., Robinson, J.P., Hirleman, E.D., Bhunia, A.K., 2007. Optical forward-scattering for detection of Listeria monocytogenes and other Listeria species. Biosensors and Bioelectronics 22, 1664–1671. https://doi.org/10.1016/j.bios.2006.07.028

Banada, P.P., Huff, K., Bae, E., Rajwa, B., Aroonnual, A., Bayraktar, B., Adil, A., Robinson, J.P., Hirleman, E.D., Bhunia, A.K., 2009. Label-free detection of multiple bacterial pathogens using light-scattering sensor. Biosens Bioelectron 24, 1685–1692. https://doi.org/10.1016/j.bios.2008.08.053

Bayraktar, B., Banada, P.P., Hirleman, E.D., Bhunia, A.K., Robinson, J.P., Rajwa, B., 2006. Feature extraction from light-scatter patterns of Listeria colonies for identification and classification. J Biomed Opt 11, 34006. https://doi.org/10.1117/1.2203987

Bhunia, A.K., Bae, E., Rajwa, B., Robinson, J.P., Hirleman, E.D., 2012. Utilization of Optical Forward Scatter Image Biological Database: Foodborne Pathogen Colony Differentiation and Detection. In Omics, Microbial Modeling and Technologies for Foodborne Pathogens, Edited by: Xianghe Yan, Vijay K. Juneja, Pina M. Fratamico and James L. Smith, Destech Publ, Lancaster, PA, pp. 553–578.

Dundar, M.M., Hirleman, E.D., Bhunia, A.K., Robinson, J.P., Rajwa, B., 2009. Learning with a non-exhaustive training dataset: a case study: detection of bacteria cultures using optical-scattering technology, in: Proceedings of the 15th ACM SIGKDD International Conference on Knowledge Discovery and Data Mining. ACM, pp. 279–288. https://dl.acm.org/citation.cfm?id=1557055

He, Y., Reed, S., Bhunia, A.K., Gehring, A., Nguyen, L.-H., Irwin, P.L., 2015. Rapid identification and classification of Campylobacter spp. using laser optical scattering technology. Food Microbiol. 47, 28–35. https://doi.org/10.1016/j.fm.2014.11.004

Huff, K., Aroonnual, A., Littlejohn, A.E.F., Rajwa, B., Bae, E., Banada, P.P., Patsekin, V., Hirleman, E.D., Robinson, J.P., Richards, G.P., Bhunia, A.K., 2012. Light-scattering sensor for real-time identification of Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae colonies on solid agar plate. Microb Biotechnol 5, 607–620. https://doi.org/10.1111/j.1751-7915.2012.00349.x

Kim, H., Rajwa, B., Bhunia, A.K., Robinson, J.P., Bae, E., 2016. Development of a multispectral light-scatter sensor for bacterial colonies. J. Biophoton https://doi.org/10.1002/jbio.201500338

Kim, H., Singh, A.K., Bhunia, A.K., Bae, E., 2014. Laser-induced speckle scatter patterns in Bacillus colonies. Front. Microbiol. 5, 537. https://doi.org/10.3389/fmicb.2014.00537

Kim, H., Doh, I.-J., Sturgis, J., Bhunia, A.K., Robinson, J.P., Bae, E., 2016. Reflected scatteromery for noninvasive interrogation of bacterial colonies. J. Biomed. Opt. 107004. http://dx.doi.org/10.1117/1.JBO.21.10.107004

Kim, H., Doh, I.-J., Bhunia, A.K., King, Galen B., Bae, E., 2015. Development of an integrated optical analyzer for characterization of growth dynamics of bacterial colonies. J. Biophoton. 6, 929-937. http://dx.doi.org/10.1002/jbio.201200224

Kim, H., Bai N., Bhunia, A.K., Hirleman, E.D., Bae, E., 2013. Scalar diffraction modeling of multispectral forward scatter patterns from bacterial colonies. Opt. Exp. 8545-8554. https://doi.org/10.1364/OE.23.008545

Rajwa, B., Dundar, M.M., Akova, F., Bettasso, A., Patsekin, V., Dan Hirleman, E., Bhunia, A.K., Robinson, J.P., 2010. Discovering the unknown: Detection of emerging pathogens using a label-free light-scattering system. Cytometry A 77A, 1103–1112. https://doi.org/10.1002/cyto.a.20978

Rajwa, B., Dundar, M.M., Akova, F., Patsekin, V., Bae, E., Tang, Y., Dietz, J.E., Hirleman, E.D., Robinson, J.P., Bhunia, A.K., 2011. Digital microbiology: detection and classification of unknown bacterial pathogens using a label-free laser light scatter-sensing system 80290C–80290C. https://doi.org/10.1117/12.884541

Singh, A.K., Bettasso, A.M., Bae, E., Rajwa, B., Dundar, M.M., Forster, M.D., Liu, L., Barrett, B., Lovchik, J., Robinson, J.P., Hirleman, E.D., Bhunia, A.K., 2014. Laser Optical Sensor, a Label-Free On-Plate Salmonella enterica Colony Detection Tool. mBio 5, e01019-13. https://doi.org/10.1128/mBio.01019-13

Singh, A.K., Sun, X., Bai, X., Kim, H., Abdalhaseib, M.U., Bae, E., Bhunia, A.K., 2015. Label-free, non-invasive light scattering sensor for rapid screening of Bacillus colonies. J. Microbiol. Methods 109, 56–66. https://doi.org/10.1016/j.mimet.2014.12.012

Tang, Y., Kim, H., Singh, A.K., Aroonnual, A., Bae, E., Rajwa, B., Fratamico, P.M., Bhunia, A.K., 2014. Light Scattering Sensor for Direct Identification of Colonies of Escherichia coli Serogroups O26, O45, O103, O111, O121, O145 and O157. PLoS ONE 9, e105272. https://doi.org/10.1371/journal.pone.0105272

Issued patents

  • US7465560B2 - System and method for rapid detection and characterization of bacterial colonies using forward light scattering

  • US8787633B2 - System and method of organism identification

Provisional patents

  • US Patent Application number 15/515,499 - Organism identification

Page updated on 2022-01-17 12:52:49 -0500