Question

You hypothesize that a group of tolerogenic cells is located in the fetal pancreas.

a. What method would you use to identify their location within the tissue?

b. What method would you use to quantify their abundance in the whole fetal pancreas based on their surface expression of protein A and Protein B and the absence of protein C?

Answer 1

a) Immunohistochemistry would be used to identify the location of tolerogenic cells within pancreas tissues. If the markers are labeled with immunofluorescence using primary and secondary antibodies, then the tolerogenic cells will most likely be found within the tissues of the fetal pancreas. In the case of this question, the antibodies would be specific to tolerogenic cells hypothesized in the pancreas of fetal.

b) Once the tolerogenic cells are present within the pancreas of fetal, Protein A, B, and C can be marked with different colored fluorescence. The cells can be sorted by using FACS and quantified. In the sequence to quantify their abundance, in the total fetal pancreas, the tissue must be disaggregated into a cellular suspension. After the formation of this suspension, flow cytometry performs to quantify their abundance based on the surface expression of Protein A and Protein B and the absence of Protein C. The difference is there would be multiple markers for the three proteins. One mark for Protein A, a different mark for Protein B and another mark for Protein C. But, Protein C is absent, there should be no emission for that mark.