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If you need to wait longer than an hour, you may need to fix the cells after step three. This can preserve them for at least several days. (This will stabilize the light scatter and inactivate most biohazardous agents).
Controls will require fixation using the same procedure. Cells should not be fixed if they need to remain viable. There are several methods available, please refer to the fixation section in the indirect staining protocol. The fixation for different antigens will require optimization by the user.