Disclaimer
Due to confidentiality agreements and out of respect for previous employers I have not gone into much detail about the in-lab specifics of my past work but would be more than happy to elaborate as much as needed with respect to my previous employers in an interview setting.
Flow Cytometry
Flow was introduced to me on the Attune NxT (invitrogen) which is a 4 laser 14 channel cytometer that I used for many in vitro and in vivo experiments with a range of cell types. I focused primarily on human PBMCs I isolated from Leukopaks and further differentiated using staining or negative magnetic isolation via STEM CELL kits. My experiments consisted of isolation and stimulation of primary immune cells of potential drug candidates. Specific experiments looked into T-cell differentiation, phosphorylation, and cytokine secretion intracellularly via protein inhibition in the golgi apparatus and in supernatant of cell cultures.
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I completed my data analysis using FlowJo and visualized this data on GraphPad Prism. All of my experiments were compiled into lab reports hosted on the company's ELN server for review by colleagues.
Cell Culture
I have worked with adherent, and suspension cell cultures ranging in volumes of 96 well plates to 50L bioreactors. To monitor the progression of these cultures I have used sample analysis machines and mass flow controllers to ensure success in the lab. I have utilized aseptic techniques to conduct growth, transduction, transfection, and lysis of cultures for DNA, and protein production purposes. My most proud work with cell culture would be an independent shake flask study where I successfully introduced a new material to the process and measured the effects it had on the process. The effects were quantified using data analysis on cell statistics such as viability, density, and changes in the nutrient and metabolites from the culture.
Protein Assay Process Development
In order to determine how much protein was in a sample, I worked with co-workers to develop a new protocol that would work best for our lab. This involved compiling protocols from published documents and combining them into our own that would successfully detect the amount of protein using the limited supplies of a startup company. Validation of this assay was done using a commercially verified assay to ensure accuracy. During this process, I was able to learn how to use a plate reader and I developed an automatic data analysis Exel Sheet that would pull values directly from the machine output into an organized and user-friendly report.
My previous lab experience has given me the opportunity to work with the following analysis instruments for daily sampling of suspension bioreactors. I completed a data bridging study with the NOVA BIO-FLEX (1 & 2) and the VI-Cell with data analysis in excel using culture samples and standards to ensure accurate data output.
