Date of Award
8-2024
Document Type
Thesis
Degree Name
Master of Science (MS)
Department
Chemical Engineering
Committee Chair/Advisor
Dr. Marc Birtwistle
Committee Member
Dr. Mark Blenner
Committee Member
Dr. David Karig
Committee Member
Dr. Adam Melvin
Abstract
Cell barcodes are capable of being used to answer many different biological questions. They have been used to track the lineage of cells to identifying the function of a gene. While there are multiple different methods to creating cell barcodes, they are limited in their scalability and application. In a previous publication we propose and computationally prove an optical single-cell barcoding method that bridges fast and scalable readouts with the benefits of genetic encoding. In this approach fluorescent proteins (fps) are combined to form fluorescent barcodes that can then be analyzed using a spectral flow cytometer. Here, we test the experimental viability of this barcoding approach at a small scale. We construct ~150 barcodes in a pooled format and then verify the makeup of the pool to validate the construction method. Spectral flow cytometry is then used to determine whether individual fluorescent proteins can be identified within single-cells, finding that most fluorescent proteins are identifiable. The fp identification results also help to verify the use of spectral flow cytometry as an analysis method capable of analyzing the barcoding approach. Current experiments are being performed to assess barcode identification within single-cells. Through these experiments we aim to show that this barcoding method is able to pair fast and scalable readouts through the use of spectral flow cytometry with genetic encoding.
Recommended Citation
Pritko, Daniel, "Genetically-Encoded Optical Barcodes for Single-Cell Analysis" (2024). All Theses. 4348.
https://open.clemson.edu/all_theses/4348