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==== Can I implement Quartz WTA in PDMS microwell arrays? ==== * Loading is a problem. For MIDAS, we load 3 ul of 10-15 cells/ul in PBS over each array. Quartz is written for 1 cell in 0.4 ul 0.5% NP-40 in a 0.2 ul low bind tube. ** I don't have to load the entire array. I could load 0.4 ul of 10-15 cells/ul into the center to avoid changing concentrations. * Quartz sorts into the lysis buffer, but I want to load and image the cells in the wells before starting. ** Imaging whole cells means that my loading buffer has to be isotonic. * Lysing is also a problem. ** MIDAS lyses with 1.5X volumes of ALS, neutralized by the same volume of NS --> 4X volume after lysis. ** Quartz lyses in 0.4 ul of 0.5% NP-40. This is also hypotonic, which helps lysis. * I could load and image in PBS, then add enough NP-40 to make 0.5%. ** Mixing would be critical, as in MIDAS ALS/NS. ** Even with 0.5% NP-40, the cells may not lyse efficiently in an isotonic solution. * The best solution may be to use the MIDAS loading and lysis protocol with scaled down volume. ** Load 0.4 ul. ** Lyse with 0.6 ul ALS ** Neutralize with 0.6 ul NS. ** Total volume = 1.6 ul.
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