The fast changing field of flow cytometry has required the flow core to replace old instrumentation and up-grade the newer instruments that we have in order to meet the requirements of our CCR investigators. The legacy MoFlo cell sorter was transfered to NEI in order for us to obtain a BD FACSAria II with a 100mW UV laser to enable core to sorttumor stem cells using the side population technique. The green lasers that were added to an anlyzer and a cell sorter is being used by numerous investigators to detect red fluorescent proteins expressed in transgenic mice and transfected or transduced cell lines. The green laser has also been used to sort lung tumor stem cells from the bulk of the tumor using the differing expression of nile red in the membrane lipids of the two cell types. More of the investigators have increased the complexity of their experiments and are routinely running experiments with 6-9 antibodies to define distinct populations of tumor infiltrating lymphocytes in different tumor models. Several linvestigators are now sorting tumor infiltrating leukocytes from various tumors in order to detect RNA expression involved in the inflammatory response to the tumor measured by quantitative RT-PCR. The larger number of fluorescence detectors has greatly increased the quality and complexity of information generated about the role of inflammation in tumor progression with each experiment. The flow core staff has trained seven investigators in the first three quarters of the year to run their own samples. The instruments are used daily by the trained investigators often into the night and on the weekends. This leaves more time for the flow core staff to sort approximately 537 samples and analyze about 6700 samples in the first three quarters of this year. Without the investigators acquiring the data and analyzing their own samples, the government would have to hire full time experienced individuals to perform the same volume of work. At least 23 papers have been published in the past year using data obtained in the flow core. In the past year, data has been generated (or samples have been sorted) for 78 individual scientists from the laboratories of 36 principal investigators. Although the CCR-Frederick Flow Cytometry Core is part of the CIP, investigators from this program account for approximately 49% of the use of this facility. The future goals include keeping up with cutting-edge technology in the flow lab by expanding the pool of investigators trained to run and analyze their own samples to all of CCR in Frederick thereby expanding the use, productivity and quality of the science generated using the core's instrumentation and expertise. This could be achieved by adding a 405 laser to the LSRII Fortessa and replacing the two BD FACScans with small 2 laser 6-color instruments.
| Gudmundsdottir, Bjorg; Gudmundsson, Kristbjorn O; Klarmann, Kimberly D et al. (2018) POGZ Is Required for Silencing Mouse Embryonic ?-like Hemoglobin and Human Fetal Hemoglobin Expression. Cell Rep 23:3236-3248 |
| Thirthagiri, Eswary; Klarmann, Kimberly D; Shukla, Anil K et al. (2016) BRCA2 minor transcript lacking exons 4-7 supports viability in mice and may account for survival of humans with a pathogenic biallelic mutation. Hum Mol Genet 25:1934-1945 |
