UNIVERSITY PARK, Pa – , at the Huck Institutes of the Life Sciences on the University Park campus, is equipped with flow cytometers and cell sorters that enable researchers to examine suspended cells within the size range of the submicron to 50 micron. One micron is one thousandth of a millimeter.
“With flow cytometry, we can understand the cellular phenotype and a number of biochemical functions on a single cell basis in high throughput manner,” explained Rajeswaran Mani, director of the facility and assistant research professor. “We can get more information from the cell such as identifying the molecules, usually antigens, that are on the surface of or within the cells. Traditionally, this has been applied to hematology and cell biology work. Now we are seeing more applications for plant biology, studying things like grasses and soil bacteria.”
Mani came to Penn State in September 2022 from the Levine Cancer Institute in North Carolina. His primary research interests are blood cancers and immune cells. During Mani’s graduate and postdoctoral research at the Ohio State University, he became interested in further using cytometry applications in his own research and exploring how it could be useful to the research of others.
Several types of techniques are utilized at Penn State’s Flow Cytometry Facility to analyze various parts and processes of cells, according to Mani. Some of these techniques include, immunophenotyping, cell cycle/DNA ploidy analysis, apoptosis assays, imaging flow cytometry, and cell sorting.
Immunophenotyping is used to identify the antigens expressed by immune cells. Cell cycle/DNA ploidy analysis is used to calculate the percentage of cell populations in the different stages of that cell’s cycle and the DNA content. Apoptosis, or cell death, assays analyze the proportion of cells in the various phases of the cell death process. Imaging flow cytometry, a combination of microscopy and flow cytometry, is used to image and analyze the characteristics of cells in rapid fashion to derive robust statistics on the image parameters, including information about shape, size, location and texture, among others.
Finally, the facility also does cell sorting, in which targeted cells (or nuclei) are identified and then separated according to their phenotype; this technique is used for establishing a pure population from a sample containing different types of cells.
The instrumentation currently available at the facility includes the BD Biosciences LSRFortessa SORP, which Mani refers as the workhorse of the facility. Fortessa has more detection channels and is equipped with High Throughput Sampler which can handle samples in plate mode, according to Mani. The facility is also home to a Beckman Coulter MoFlo Astrios EQ Cell Sorter, housed in a bio-safety cabinet to handle BSL-2 materials. This flow sorter can also deposit “single-cell” into a collection well for single cell cloning applications.
Mani, who came to Penn State with an interest in supporting the next generation of scientists, said he is passionate about furthering the reach of the Flow Cytometry Facility to support more research, collaborations, and to increase the resources available for immunobiology.
“With our unique set up at the Huck, we are poised to support multi-disciplinary research happening across the colleges,” Mani added.
The services of the facility are available to all Penn State departments and to researchers outside of the University. The facilities’ faculty and staff, including research technologist, Desa Rae Abrams, are available to help researchers design their experiments and protocols.
“Anything that has to do with cells in suspension phase can be analyzed with flow cytometry,” Mani said.
