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. Author manuscript; available in PMC: 2018 May 23.
Published in final edited form as: Nat Protoc. 2017 Jul 20;12(8):1620–1638. doi: 10.1038/nprot.2017.059

TABLE 2.

Troubleshooting table.

Step Problem Possible reason Solution
2 Weight loss of the polymer (incorrect balancing) Static electricity Use a static gun (e.g., Milty Zerostat 3 Anti-Static Gun)
20 Air bubble in the gels Unsuitable pipette was used Use a positive-displacement pipette
22A(xiii), 23B(i) Cells sink to the bottom Preincubation time is too short Increase the preincubation time
23A(i) Inability to obtain clear images of the 96-well plate A long-working-distance objective was not used Use a long-working-distance objective or switch to a glass-bottom 96-well plate
23D(i) Cell structures collapse Fixation conditions are too harsh Decrease PFA concentration (from 3.7 to 2%) or decrease the PFA incubation time
23D(iv) Poor image quality Permeabilization conditions are too harsh Decrease Triton X-100 concentration (to between 0.1 and 1% ((vol/vol))) or decrease Triton X-100 incubation time (to between 10 and 20 min)