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pubmed.ncbi.nlm.nih.gov/7054160 Irobi E, Aguda AH, Larsson M, et al

The scan numbers tend to catch up as the protocol continues past the initial three cycles

Koleini, N., Nickel, B

After fixing with 4% paraformaldehyde, the cells were exposed to cold methanol for 1 min and then treated with 1 N HCl at RT for 1 h before performing immunocytochemistry