As soon as you put down the alcohol bottle you should

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As soon as you put down the Alcohol Bottle you should pick up your Water Bottle and start rinsing. 13. Tip the Slide to allow excess Water to fall into the Sink. If you ʼ re using Controls, repeat the Decolorization for the other 2 Smears. 14. Flood the Smears with Safranin (also spelled Safranine) and let your Slide sit on the Staining Rack for 1 Minute. 15. At the End of the Minute, tip the Slide to allow Safranin to drain into your Sink. 16. Rinse off the remaining Safranin with DI Water. 17. Tip the Slide to allow excess Water to drain into your Sink. 18. Dry the Slide using Blotting Paper. 19. Check one of the Smears under 10X. When it’s In Focus, go on to the 100X and Immersion Oil. You cannot correctly determine the Gram status of a Smear unless you are using the 100X Lens with Immersion Oil.
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Lab 3 Page 10 Gram Stain Check List By the end of the Term, you ʼ ll be able to perform Gram Stains without referring to the Lab Manual. And you ʼ ll screw them up royally, too. This is why Pilots and Surgeons always use a Check List. So here ʼ s our Gram Stain Check List: • Make a Smear • Air dry • Heat fix • Crystal Violet: 1 Minute • Rinse. • Iodine: 1 Minute • Rinse • Ethanol: 10 Drops, 10 Seconds • Rinse • Safranin: 1 Minute • Rinse • Blot dry Handy-Dandy Mnemonic (L= mindful) : Craving Indian? Eat Samosas! (Crystal Violet, Iodine, Ethanol, Safranin) ________________________________________________________________________________ “This baby belongs to them cloners. What you got here is a Kamino Saberdart.” I wonder why it didn't show up in the analysis archives?’ It's these funny little cuts on the side that give it away. Those analysis droids only focus on symbols. Huh! I should think that you Jedi would have more respect for the difference between knowledge and... heh heh heh... wisdom.” Well if droids could think, there'd be none of us here, would there? These Kaminos, are they friendly?” That depends.’ Depends on what, Dex?” On how well your manners are, and how big your pocketbook is.’ © 2002 LucasFilm Ltd.
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Lab 3 Page 11 Gram Stain Troubleshooting There are a whole lot of Points tied-up in Gram Stains in MIC 101 so your Ability to reliably perform Gram Stains is an important Survival Skill. There are three common Mistakes made when doing Gram Stains. 1. Smears that are too Thick This is by far the most common Problem with Gram Stains. Do not fall in love with your Smears. Be extremely critical. If you’ve got crapola Smears, make new Smears. It doesn’t take that long. Be sure you make Smears that are very Light. You want to be viewing individually stained Cells, not Crowds of Cells that can collect Crystal Violet-Iodine Complex or Safranin in between. 2. Uneven or inconsistent Decolorization This is another Trouble Spot. It’s a bit tedious decolorizing each Smear one at a Time and there’s a Tendency to get sloppy.
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