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Another decolorization method is to simply flood the

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Another Decolorization Method is to simply flood the Slide with Ethanol for 10-15 Seconds and then immediately rinse the Slide with DI Water. 3. Forgetting to Heat Fix This is surprisingly easy to do. And if you forget to Heat Fix, all of your Bacteria do a Roaring Rapids down the Sink during your first Rinse. Get into the Habit of Heat Fixing all of your Smears at the same Time whenever this is Practical. 4. Heat Fixing before your Smear has completely Dried A watched Pot never boils and a watched Smear never dries. Make your Smears as far in Advance as is Reasonable. You can go into Geisha Mode and fan your Slides with a Folder but this isn’t going to make a huge Difference. Try and use as little Water as possible while making your Smears. __________________________________________________________ If an item does not appear in our records, it does not exist. © 2002 LucasFilm Ltd.
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Lab 3 Page 12 Viewing a Gram Stain Place your Slide (Smear-Side Up) on your Stage. If you can’t remember which Side has the Smear, hold the Slide at an Angle such that you can reflect either Window Light or a Fluorescent Light off the Slide; you’ll see a Smudge on the Slide with the Smear. If you place your Slide on the Stage Upside-Down, you ʼ ll still be able to focus on the Smear using the 10X (Yellow) Objective. But you won ʼ t be able to see anything on the 40X (Blue) Objective because the Working Distance of the 40X (Blue) Objective is less than the Thickness of our Slides. Remember this if you can get your Bacteria In Focus on the 10X (Yellow) Objective but can ʼ t get them In Focus on the 40X (Blue) Objective. If you can focus on 10X but not on 40X, your Slide is Upside-Down! Everyone in Lab will point at you and laugh. Can ʼ t focus? Go back to your “P” Slide. Try to locate and focus your Bacteria using your 10X (Yellow) Objective. Remember, your Scope is Parfocal. Once it’s in Focus for the 10X (Yellow) Lens, it should be in Focus for the 40X (Blue) Lens. And once it’s In Focus for any one Slide, it should be In Focus for any Slides that follow. You’ll swear there’s Nothing there except some tiny Pink Specks. Well, those are the Bacteria. And E. coli is one of the larger Bacteria, too. This is why MIC 101 is called Microbiology. For the most part, we’ll be looking at Stuff this small. After focusing on 10X, move up to 40X and readjust the Fine Focus and re-center the Smear if necessary. In order to see any Detail at all on your Bacteria you’ll need to use the 100X (White) Lens. This is a special Lens designed to be used with Immersion Oil. And that’s covered on the next Page... ___________________________________________________________________________________ Dangerous and disturbing this puzzle is. Only a Jedi could have erased those files. But who, and why, harder to answer. Meditate on this I will. © 2002 LucasFilm Ltd.
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Lab 3 Page 13 Using Oil Immersion You’ll need to go up to the 100X Oil Immersion Lens to learn anything about the Morphology of your Bacteria. Using Oil Immersion is quite straightforward once you get the Hang of it.
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