Development Of An Acoustofluidic Fluorescence Activated Cell Sorter (facs)
Open Access
- Author:
- Nawaz, Ahmad Ahsan
- Graduate Program:
- Engineering Science
- Degree:
- Doctor of Philosophy
- Document Type:
- Dissertation
- Date of Defense:
- December 18, 2014
- Committee Members:
- Jun Huang, Dissertation Advisor/Co-Advisor
Jun Huang, Committee Chair/Co-Chair
Corina Stefania Drapaca, Committee Member
Bruce Gluckman, Committee Member
Bernhard R Tittmann, Committee Member
Siyang Zheng, Committee Member - Keywords:
- Microfluidic flow cytometry
3D focusing
cell sorter
hydrodynamic
aerosol free sorting. - Abstract:
- This dissertation details the development of a miniature, low cost, and high-throughput fluorescence-activated cell sorter (FACS). This miniature FACS device contains the essential functionalities that exist in bulky, expensive and complex commercially available FACS machines, while being significantly more compact, inexpensive, and biosafe. The first stage of the dissertation involves development and optimization of a “microfluidic drifting” cell/particle focusing unit that can achieve 3D alignment of cells/particle flowing through the device with submicron precision. The focused file of flowing particles analyzed by illuminating a laser light and collecting scattering signal from the particles demonstrated a 2.37% coefficient of variation (CV). Results as good as commercial flow cytometer were obtained when analyzing HEK-293 cells. Moreover, well separated eight populations were discerned when subjected to stringent 8-peak rainbow calibration test indicating high detection sensitivity of the cytometry device. Next stage entails expanding the microfluidic flow cytometry device from single parameter to a multi-parametric, optical fiber integrated flow cytometry device. Immunological analysis of human blood and CD4+ counting delineated capabilities of the device to achieve similar results to large bench-top commercial flow cytometer. Following step was involved the development of an acoustofluidic fluorescence activated cell sorting (FACS) device that simultaneously performs high-throughput, high-resolution biological studies and standing surface acoustic wave (SSAW) based cell sorting, integrated onto a single chip. This FACS device utilizes the “microfluidic drifting” phenomenon to pre-focus cells/particles followed by SSAWs to sort cells of desired biological characteristics. At a throughput of ~ 1200 cells/s, HeLa cells were sorted viably at a purity of 92.3 ± 3.39 %. The final part of the dissertation sheds light upon future directions and shows some preliminary results elucidating the capability of the acoustic FACS as a hematology analyzer, its potential to analyze induced human sputum and analyze and sort large organism sorting such as c. elegans. The development our acoustofluidic FACS is a leap towards realization of a high-resolution, low-cost miniature FACS. Through this dissertation, great efforts have been extended towards achieving this objective.