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Tip #381. Backgating for confident FACS analysis.
Not sure if your "positive" staining is real or is due to contamination by dead cells or autofluorescent cells?
For an excellent example of how to use stringent gating, and intelligent backgating, to eliminate false positive staining see Vijaynand et al., NEJM (2007) 356:1410. See especially supplemental Figure E3 which shows you step by step how dead cells, doublets, and macrophages contribute to false PE/FITC staining in a tetramer analysis of airway NKT cells.
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Ee Wern Su & Kristy Pilbeam
Graduate student and Technician III, respectively
Department of Immunology
On global medicine... |
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1. What inspired you to pursue a career in the medical sciences?
EWS: To be in the position to contribute towards the betterment of global healthcare and advancement of medical therapies through research.
KP: My aspirations to serve the global community in health care is similar to Wern’s. However, I would like to focus my efforts towards providing quality healthcare for underserved communities as a physician.
2. Having experienced/been exposed to health care systems in other countries, what do you consider to be the biggest issue facing US medical care?
EWS (Malaysian citizen): The opportunity for citizens to seek and practitioners to provide, quality healthcare at a fair price - variables that are inextricably linked and dictated by the cost of health insurance.
KP (expertise in Chilean health care): The rising cost of medical malpractice insurance, which drives physicians out of certain states and from primary care service. In addition, we are plagued by a shortage in all types of healthcare workers and rising healthcare cost that doesn’t seem to be improving.
3. When you are out for coffee at the Strip, what are the sociopolitical issues you talk about? What concerns people of your generation?
EWS: The search for a strong political identity that can satisfy one’s personal beliefs in a time where borders between cultures are rapidly blurring.
KP: At this point in time international politics is a hot topic (which is my big interest), but often discussions touch on our disappointment with the education and health systems. More specifically, how there is little effort and resources devoted to improving the several problems faced by these systems.
4. And if you are at a wine bar?
EWS & KP: We both concur that Pennsylvania’s legislation regulating the import and export of state and foreign wines severely limits our wine selection. In addition, we are constantly bothered by the poor bar manners of fellow patrons.
5. Say, where are these good cafes and wine bars anyway?
EWS & KP: Given that there aren’t many wine bars in Pittsburgh, these are a few places worth patronizing.
La Prima – Corner of Penn Ave in the Strip
Taza de oro – Cute place for coffee on Highland
Fat Heads – Food and beer here will make up for the short list of good wine bars.
Claddagh – Once again, good beer and environment.
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