Detection of Leishmania Parasites via Flow Cytometry
Document Type
Oral Presentation
Campus where you would like to present
SURC 140
Start Date
21-5-2015
End Date
21-5-2015
Keywords
Leishmania, Flow Cytometry, Macrophage Infection
Abstract
Leishmania species cause a spectrum of diseases collectively referred to as the Leishmaniases. These unicellular parasites generate 1.3 million new cases of infection worldwide as well as cause an approximated 20,000 to 30,000 deaths annually. Current techniques for studying Leishmaniasis in laboratory animals require laborious serial plating of dilution series, incubations of samples for weeks, followed by microscopic screening of each individual well in 96 well plates. Through the generous funding of both the W. M. Keck Foundation and the Murdock Fund, Biological Sciences recently obtained a flow cytometer. This study aimed at assessing if flow cytometry could differentiate cells infected with Leishmania parasites from uninfected cells. Two species of the parasite, L. infantum and L. major, were stained with an antibody conjugated to a florescent fluorochrome, using both flow cytometry and photomicroscopy to detect the parasites. Fluorescent staining of parasites illustrated a shift in florescence intensity using flow cytometry and easily visualized parasites using microscopy. However, a modified protocol for intracellular staining of the murine macrophage cell line, J774A.1, did not reveal a separation between the infected cells and uninfected cells using flow cytometry. The parasites within cells were also undetectable using fluorescent microscopy. Although the hypothesis that there would be a detectable segregation between infected and uninfected cells was not met, this work will continue to attempt to develop what would be a time saving tool in Leishmania research.
Recommended Citation
Wenger, Analiese, "Detection of Leishmania Parasites via Flow Cytometry" (2015). Symposium Of University Research and Creative Expression (SOURCE). 98.
https://digitalcommons.cwu.edu/source/2015/oralpresentations/98
Department/Program
Biological Sciences
Additional Mentoring Department
Biological Sciences
Additional Mentoring Department
Biological Sciences
Detection of Leishmania Parasites via Flow Cytometry
SURC 140
Leishmania species cause a spectrum of diseases collectively referred to as the Leishmaniases. These unicellular parasites generate 1.3 million new cases of infection worldwide as well as cause an approximated 20,000 to 30,000 deaths annually. Current techniques for studying Leishmaniasis in laboratory animals require laborious serial plating of dilution series, incubations of samples for weeks, followed by microscopic screening of each individual well in 96 well plates. Through the generous funding of both the W. M. Keck Foundation and the Murdock Fund, Biological Sciences recently obtained a flow cytometer. This study aimed at assessing if flow cytometry could differentiate cells infected with Leishmania parasites from uninfected cells. Two species of the parasite, L. infantum and L. major, were stained with an antibody conjugated to a florescent fluorochrome, using both flow cytometry and photomicroscopy to detect the parasites. Fluorescent staining of parasites illustrated a shift in florescence intensity using flow cytometry and easily visualized parasites using microscopy. However, a modified protocol for intracellular staining of the murine macrophage cell line, J774A.1, did not reveal a separation between the infected cells and uninfected cells using flow cytometry. The parasites within cells were also undetectable using fluorescent microscopy. Although the hypothesis that there would be a detectable segregation between infected and uninfected cells was not met, this work will continue to attempt to develop what would be a time saving tool in Leishmania research.
Faculty Mentor(s)
Gabrielle Stryker, Blaise Dondji