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.

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