ORCID
https://orcid.org/0000-0001-5879-2344
Date of Award
Winter 2025
Language
English
Embargo Period
1-17-2025
Document Type
Master's Thesis
Degree Name
Master of Science (MS)
College/School/Department
Department of Chemistry
Program
Chemistry
First Advisor
Jun Wang
Committee Members
Alan Chen, Michael Yeung
Keywords
photolithography, microbeads, Janus, MIST, cytokine, microchip
Subject Categories
Chemistry
Abstract
To better advance our knowledge of cellular behavior, development of an organism, and disease mechanisms understanding cellular heterogeneity in the functional proteome is essential. Bulk measurements across a cell population often fail to capture the variations which exist between individual cells, however single cell proteomic technology has increasingly demonstrated the characterization of cellular heterogeneity. A single cell functional proteomics assay needs to be both high-throughput and multiplexed to effectively identify biomarkers and provide a quantitative correlation to better understand the interrelationships within complex protein networks. Janus particles have the ability to be engineered to have good compatibility and stability in a complex biological environment. Functionalization or surface modification of a microparticle allows for a precise targeted detection of cytokines. The dual nature of a Janus particle allows for both surface interaction and detection in a cellular environment. In this first project a Janus particle is fabricated to demonstrate the asymmetric patterning and evaluate its capabilities in detecting cytokines in HL-60 cells. Fluorescent microscopy imaging confirms the Janus particle design and the ability to attach to a cell membrane, however the experimental yield is low for an adequate quantitative analysis with cytokines. In the second project a Multiplex In Situ Tagging (MIST) array is designed and the fluorescence ELISA sandwich technique is used to quantify the release of cytokines (P53, Total COX-2, TNF RI, Phospho-GSK-3, Cleaved Caspace-3, and Phospho-CREB) in THP1 cells. On an assembled microchip a multiplex DNA encoded microbead array with microwells has a high sensitivity to measure cytokine expression. The MIST array demonstrated shows the ability for a multiplex cytokine detection, however sensitivity demonstrated in this experiment would need to be optimized for a better understanding of the cellular heterogeneity of the cytokines expressed in the single cells.
License
This work is licensed under the University at Albany Standard Author Agreement.
Recommended Citation
George, Justin, "Fabrication of Functional Microparticles for Biological Analysis" (2025). Electronic Theses & Dissertations (2024 - present). 92.
https://scholarsarchive.library.albany.edu/etd/92
