We introduce a unique microfluidic-based approach for the high-throughput non-destructive assaying of cells without the need for specific labels or reagents. Based on measurement of both static and dynamic cell mechanical properties using applied optical forces, we will apply this technique (known as """"""""optical stretching"""""""") in a high-speed high-throughput manner. To date, optical stretching has been used only on small cell numbers;however, high- intensity, microscale laser sources and the integration of these within dynamic microfluidic systems has enabled our proposed approach. In this, fully integrated optical-based sensors and mechanical stretchers will be used to identify and, upon demand, isolate single cells. Once identified, such targeted cells can then be transported on-chip to culture chambers within the device or for dispensing into standard bio-laboratory instrumentation for off-chip analysis. Though there is broad need, our proposed technology will be tested and developed using malaria parasite infected red blood cells as the target cell. This work will be done in collaboration with the Laboratory of Malaria and Vector Research at the NIAID.
Our aims i nclude:
Aim 1 : Mechanical Property Detection and Interpretation. We will employ optical manipulation methods integrated within microfluidic systems for label-free, non-destructive cell mechanical property measurement. Modeling approaches will be developed for both interpretation of applied force/deformation experimental data and for device design. Here, malaria-infected red blood cells will provide a good model target since cell stiffness changes dramatically during parasite development. Demonstrating greatly simplified device designs and associated ease-of-use, we will install an instrument in an active NIH laboratory.
Aim 2 : Optical Manipulation for Cell Identification and Isolation. We will integrate optical methods within microfluidic systems for single cell detection and manipulation. Here, methods for both on-chip cell isolation and off-chip isolation will be developed and used to improve our installed NIH protototype.
Aim 3 : High Throughput Mechanical Testing. To achieve high-throughputs, modified microfluidic and faster detection techniques will be required. In this phase, the coupling of hydrodynamic and optical forces will be explored to improve device performance. In addition, time-varying optical forces will be employed to identify optimal signal response and dynamic physical properties.

Public Health Relevance

We propose to develop new methods based on physical property measurement for the high-throughput analysis of cells. Such techniques that avoid the need for labels can be not only simpler and less expensive, they can be less harmful to the cell for applications where cell viability post-assaying is desirable.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI079347-01A1
Application #
7736282
Study Section
Instrumentation and Systems Development Study Section (ISD)
Program Officer
Wali, Tonu M
Project Start
2009-08-15
Project End
2013-07-31
Budget Start
2009-08-15
Budget End
2010-07-31
Support Year
1
Fiscal Year
2009
Total Cost
Indirect Cost
Name
Colorado School of Mines
Department
Engineering (All Types)
Type
Schools of Arts and Sciences
DUNS #
010628170
City
Golden
State
CO
Country
United States
Zip Code
80401
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Roth, Kevin B; Eggleton, Charles D; Neeves, Keith B et al. (2013) Measuring cell mechanics by optical alignment compression cytometry. Lab Chip 13:1571-7
Sawetzki, Tobias; Eggleton, Charles D; Desai, Sanjay A et al. (2013) Viscoelasticity as a biomarker for high-throughput flow cytometry. Biophys J 105:2281-8
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Sraj, Ihab; Szatmary, Alex C; Desai, Sanjay A et al. (2012) Erythrocyte deformation in high-throughput optical stretchers. Phys Rev E Stat Nonlin Soft Matter Phys 85:041923
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Sraj, Ihab; Szatmary, Alex C; Marr, David W M et al. (2010) Dynamic ray tracing for modeling optical cell manipulation. Opt Express 18:16702-14