BD Accuri News

Melanie Gubbels Bupp on Education

Melanie Gubbels Bupp, PhD, is an immunologist and assistant professor of biology at Randolph-Macon College in Ashland, VA, a private liberal arts and sciences college with an enrollment of 1,200 undergraduates. Dr. Gubbels Bupp told us about the key role her BD Accuri™ C6 flow cytometer plays in both her teaching and research, how it’s helpful to students, and which Star Wars character it resembles.


Q: Would you tell us about how you use flow cytometry in education? What is your goal in including it in the curriculum?

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Figure 1. Flow cytometry immunophenotyping lab in an undergraduate immunology course
 
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Flow cytometry immunophenotyping lab in an undergraduate immunology course

In Melanie Gubbels Bupp's undergraduate immunology lab at Randolph-Macon College, students perform a three-lab experiment on leucocyte subset infiltration during thioglycollate-induced peritonitis.

  • Lab Session I:
  • Students obtain time zero blood differentials on mice by blood smear microscopy.
  • The instructor injects mice with thioglycollate to induce peritonitis.
  • Lab Session II:
  • Students collect peripheral blood from mice and perform peritoneal lavage to obtain infiltrating cells.
  • Students obtain blood differentials on peripheral blood smears.
  • Students perform cell staining with an antibody panel designed to differentiate various white blood cell (WBC) populations (see figure).
  • Cells are fixed and held for up to 1 week, covered and refrigerated.
  • Lab Session III:
  • Students collect data on the BD Accuri™ C6 flow cytometer using an instructor-created template.
  • The instructor uploads FC plots and population percentage information to the course website.
  • Students calculate and graph the WBC population numbers with time for both the peripheral blood and peritoneal lavage data.

Dr. Gubbels Bupp: I use the BD Accuri C6 in the lab portion of my undergraduate immunology course, because I want my students to perform meaningful experiments with the actual tools that immunologists use. They do at least two multi-week experiments that involve flow cytometry. The first (Figure 1) involves inducing peritonitis in mice with thioglycollate. Students then monitor white blood cell populations at various time points in the blood via manual blood differentials and within the peritoneal cavity using the BD Accuri C6 and antibodies to macrophages, neutrophils, T cells, and B cells.

For the second experiment, students research and propose a substance that might enhance or inhibit phagocytosis. Then they ascertain how that substance affects the ability of human macrophages (THP-1 cells treated with PMA) to phagocytose FITC-latex beads.

Q: Why did you choose the BD Accuri C6?

Dr. Gubbels Bupp: To be honest, it was the only cytometer I could afford! But now that I have it, I am glad that it's so easy to use. I can train undergraduates on it quickly and easily.

Q: Do you use the BD Accuri C6 in your research as well?

Dr. Gubbels Bupp: Yes, I use the cytometer in my research also. It's been a great tool for me. The work is done exclusively with undergraduates and all of them use the cytometer.

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Figure 2. Effects of calorie restriction on T-cell death in mice
 
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Effects of calorie restriction on T-cell death in mice

Mice were fed ad libitum (AL) or a 35% calorie-restricted (CR) diet for 6 weeks. T cells were isolated, stained with Annexin V and 7-AAD to assess cell death, and acquired and analyzed on the BD Accuri C6. Graphs display means ±SEM. A. Death rates of naïve CD8+ T cells were higher in AL than in CR mice (p = 0.002, N = 10–11 mice/group). B. Treatment with IL-7 dramatically reduced CD8+ T-cell death rates in AL mice but not in CR mice (p = 0.0001, N = 3–5 mice/group). Data courtesy of Melanie Gubbels Bupp, Randolph-Macon College, Ashland, VA.

Our lab is investigating the established finding that reducing caloric intake by at least 30% (calorie restriction, or CR) significantly extends the lifespan of non-human primates, rodents, and other organisms. Recent studies in our lab have used the BD Accuri C6 to explore the effects of calorie restriction on T cells. Two of my undergraduate research students, Whitney Edwards (R-MC '12) and Seth Litvin (R-MC '13), found that a 35% calorie restriction lasting 6 weeks was associated with a 3-fold reduction of naïve CD8+ T-cell death in vitro compared to cells isolated from mice fed ad libitum (AL) (Figure 2; p = 0.0002 by unpaired t-test).

We hypothesized that calorie restriction was inhibiting the "death-by-neglect" pathway. If so, then the addition of IL-7 should rescue AL T cells, but have relatively little effect on CR T cells. Litvin confirmed this, finding that IL-7 treated AL T cells experienced almost two-thirds less cell death than cells cultured without IL-7, while IL-7 had almost no effect on CR T cell death (p = 0.0001 by 2-way ANOVA).

Q: What are your students' reactions to the BD Accuri C6?

Dr. Gubbels Bupp: They are excited to work with a current instrument. They like its color and think it sounds like R2D2.

I also think it's helpful for those students who decide to go on to graduate school in immunology—or to medical school or industry, for that matter. It's pretty impressive to admissions committees to see that students are using current, applicable technology, and it gives them an advantage if they already have some experience with such a common tool in the field. And once they get into their rotation labs, it's one less thing they have to learn when they're learning so many other things.

Read the March 2013 BD Accuri™ News feature on personal flow cytometry in education »

Download the white paper, Sowing the Seeds of Knowledge: The Integration of Practical Flow Cytometry into Undergraduate Education »



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