Presenter: Emily Marie Larkin
Faculty Sponsor: Lillian Fritz-Laylin
School: UMass Amherst
Research Area: Biological Organisms
Session: Poster Session 6, 4:15 PM - 5:00 PM, Auditorium, A60
ABSTRACT
In the past 20 years, media coverage of "brain-eating" amoeba cases has increased. Naegleria fowleri, the “brain-eating” amoeba, causes primary amoebic meningoencephalitis (PAM), a rare but fatal disease characterized by the rapid proliferation of the amoeba in the brain. Despite a 97% fatality rate, there are currently few effective therapies to treat PAM due to a lack of effective treatment methods and lack of timely diagnosis. Gaining a better understanding of fundamental cell biological processes in Naegleria, such as cell division and cell motility, could identify potential avenues for new drug targets and could highlight new methods for identifying PAM. Most current research has utilized genomics and fixed cell imaging to investigate cellular processes in Naegleria. However, there are limitations to the data that can be acquired from fixed cell imaging. Developing a protocol using microscopy and live cell dyes can provide more information on dynamic processes. At present, there is insufficient understanding of optimal conditions for live cell imaging in Naegleria. To fill this gap in knowledge, we conducted experiments testing illumination conditions, different cell dyes, and at different dye concentrations to determine the cellular viabilities of non-pathogenic Naegleria gruberi using multiple measurement approaches. We found that the nucleic acid dye, Hoechst 33342, labeled the nucleus and had the highest percent viability compared to other nucleic acid dyes. We determined that a 1:2,000 Hoechst 33342 dye solution effectively distinguishes cellular division in an amoeba population. Utilizing the developed protocol, we can begin to learn more about Naegleria mitosis which could be used to identify targets for treatment against PAM.
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