bis_104_pq_11_ans_spring_08

bis_104_pq_11_ans_spring_08 - the other cell type Both...

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BIS 104 Practice Question #11 08 11. After separating Schwann cells from neurons by the FACS technique, Mukouyama attempted to verify the purity of these cell populations by reverse transcriptase PCR / northern blotting (fig. 6, panel D) and by immunofluorescence (fig. 7 a,b,c,d). Are you convinced that the FACS procedure successfully separated Schwann cells and neurons into two highly purified populations? Explain the basis for your answer. a) Using RT-PCR they probed both pure populations for cell-specific mRNA. mRNA for SG10 = neuron-specific mRNA for GFAP = Schwann cell-specific In both cases, neither cell population was expressing detectable levels of mRNA characteristic of
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Unformatted text preview: the other cell type. Both populations were shown to express mRNA for VEGF and for HGPRT as expected. b) The PCR –notherern blot results were confirmed by examining the two populations purified by FACS by immunocytochemistry. Fluoresently labeled antibodies for both cell types were applied to both populations. Examination by fluorescence microscopy showed that the populations were > 95% “pure”, that is, little or no cross-contamination of cell types. So, yes, the FACS technique successfully separated neurons from Schwann cells in this experiment....
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