Dr. Shashaanka P. Ashili Profile

Dr. Shashaanka P. Ashili

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Optical Physicist and Multidisciplinary Engineer

SPIE Journal Paper | 5 April 2012 Open Access

Method for physiologic phenotype characterization at the single-cell level in non-interacting and interacting cells

Laimonas Kelbauskas, Shashanka Ashili, Jeff Houkal, Dean Smith, Aida Mohammadreza, Kristen Lee, Jessica Forrester, Ashok Kumar, Cody Youngbull, Yanqing Tian, Mark Holl, Roger Johnson, Deirdre Meldrum, Yasser Anis, Thomas Paulson

JBO, Vol. 17, Issue 03, 037008, (April 2012) https://doi.org/10.1117/12.10.1117/1.JBO.17.3.037008

KEYWORDS: Oxygen, Optical character recognition, Sensors, Optical sensors, Microscopes, Calibration, Silica, Luminescence, Data analysis, Sensor calibration

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Proceedings Article | 15 February 2011 Paper

Automated platform for multiparameter stimulus response studies of metabolic activity at the single-cell level

Shashanka Ashili, Laimonas Kelbauskas, Jeff Houkal, Dean Smith, Yanqing Tian, Cody Youngbull, Haixin Zhu, Yasser Anis, Michael Hupp, Kristen Lee, Ashok Kumar, Juan Vela, Andrew Shabilla, Roger Johnson, Mark Holl, Deirdre Meldrum

Proceedings Volume 7929, 79290S (2011) https://doi.org/10.1117/12.875438

KEYWORDS: Sensors, Oxygen, Microfluidics, Control systems, Interfaces, Glasses, Confocal microscopy, Human-machine interfaces, Data acquisition, Motion controllers

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We have developed a fully automated platform for multiparameter characterization of physiological response of individual and small numbers of interacting cells. The platform allows for minimally invasive monitoring of cell phenotypes while administering a variety of physiological insults and stimuli by means of precisely controlled microfluidic subsystems. It features the capability to integrate a variety of sensitive intra- and extra-cellular fluorescent probes for monitoring minute intra- and extra-cellular physiological changes. The platform allows for performance of other, post- measurement analyses of individual cells such as transcriptomics. Our method is based on the measurement of extracellular metabolite concentrations in hermetically sealed ~200-pL microchambers, each containing a single cell or a small number of cells. The major components of the system are a) a confocal laser scan head to excite and detect with single photon sensitivity the emitted photons from sensors; b) a microfluidic cassette to confine and incubate individual cells, providing for dynamic application of external stimuli, and c) an integration module consisting of software and hardware for automated cassette manipulation, environmental control and data collection. The custom-built confocal scan head allows for fluorescence intensity detection with high sensitivity and spatial confinement of the excitation light to individual pixels of the sensor area, thus minimizing any phototoxic effects. The platform is designed to permit incorporation of multiple optical sensors for simultaneous detection of various metabolites of interest. The modular detector structure allows for several imaging modalities, including high resolution intracellular probe imaging and extracellular sensor readout. The integrated system allows for simulation of physiologically relevant microenvironmental stimuli and simultaneous measurement of the elicited phenotypes. We present details of system design, system characterization and metabolic response analysis of individual eukaryotic cells.

Proceedings Article | 11 February 2011 Paper

A novel method for multiparameter physiological phenotype characterization at the single-cell level

Laimonas Kelbauskas, Shashanka Ashili, Jeff Houkal, Dean Smith, Aida Mohammadreza, Kristen Lee, Ashok Kumar, Yasser Anis, Tom Paulson, Cody Youngbull, Yanqing Tian, Roger Johnson, Mark Holl, Deirdre Meldrum

Proceedings Volume 7902, 79021Q (2011) https://doi.org/10.1117/12.875483

KEYWORDS: Oxygen, Optical character recognition, Sensors, Optical sensors, Microscopes, Cancer, Microfluidics, Calibration, Data acquisition, Cell mechanics

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