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I am unaware of an optimal time for fixing gram stained smears. Heating on a slide warmer at 60C for at least 10 minutes is recommended (Bailey and Scott, 9th edition, 1994, page 68). A better method may be methanol fixing the slide (Manual of Clin Microbiology, 8th ed, 2003). However, be aware that unless the slides are heated adequately, certain organisms may still survive (MTB for one). Published temperature and times to adequately heat slides for Tb are 65-70C for 2hours or 80C for 15 min (Bailey and Scott, 9th edition, 1994, page 605).
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