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About Science Prof Online PowerPoint Resources • Science Prof Online (SPO) is a free science education website that provides fully-developed Virtual Science Classrooms, science-related PowerPoints, articles and images. The site is designed to be a helpful resource for students, educators, and anyone interested in learning about science. • The SPO Virtual Classrooms offer many educational resources, including practice test questions, review questions, lecture PowerPoints, video tutorials, sample assignments and course syllabi. New materials are continually being developed, so check back frequently, or follow us on Facebook (Science Prof Online) or Twitter (ScienceProfSPO) for updates. • Many SPO PowerPoints are available in a variety of formats, such as fully editable PowerPoint files, as well as uneditable versions in smaller file sizes, such as PowerPoint Shows and Portable Document Format (.pdf), for ease of printing. • Images used on this resource, and on the SPO website are, wherever possible, credited and linked to their source. Any words underlined and appearing in blue are links that can be clicked on for more information. PowerPoints must be viewed in slide show mode to use the hyperlinks directly. • Several helpful links to fun and interactive learning tools are included throughout the PPT and on the Smart Links slide, near the end of each presentation. You must be in slide show mode to utilize hyperlinks and animations. • This digital resource is licensed under Creative Commons Attribution-ShareAlike 3.0: • http://creativecommons.org/licenses/by-sa/3.0/ Tami Port, MS Creator of Science Prof Online Chief Executive Nerd Science Prof Online Online Education Resources, LLC info@scienceprofonline.com Alicia Cepaitis, MS Chief Creative Nerd Science Prof Online Online Education Resources, LLC alicia@scienceprofonline.com From the Virtual Microbiology Classroom on ScienceProfOnline.com Image: Compound microscope objectives, T. Port
Laboratory Exercise 1bHow to Use a Compound MicroscopeViewing a Simple Stain of Bacteria Under Oil Immersion Aren’t They Cute? ** Please plug in your microincinerators ** From the Virtual Microbiology Classroom on ScienceProfOnline.com Image: Simple stain of Staphylococcus, T. Port
Low Power Objective Lens • Has yellow band around it. • Magnifies objects 10x. • Q: What is the Total Magnification? ____ TM • Start with this lens when looking at a bacterial smear. • Q: What does the term parfocal mean? From the Virtual Microbiology Classroom on ScienceProfOnline.com Image: Microscope objective lenses, T. Port
Practice Safe Microscopy!Protect Your High-Dry Lens • Before putting any oil on your slide, make sure that the hi-dry, blue banded lens is covered up with a finger cot! From the Virtual Microbiology Classroom on ScienceProfOnline.com Image: Microscope objective lenses, T. Port
Oil Immersion Objective Lens • Has B&W band around it. • Magnifies objects 100x. • Q: What is the Total Magnification? ____ TM • Make sure that your bacterial smear is clearly in focus at 100xTM. • Put a drop of immersion oil directly on each of the three bacterial smears on your slide, then switch to the oil immersion lens. From the Virtual Microbiology Classroom on ScienceProfOnline.com Image: Microscope objective lenses, T. Port
Observing bacteria under oil immersion: • Don’t EVER use coarse focus when working with high dry or oil immersion. • Remember PARFOCAL! • Using oil immersion: • View bacteria with low power 100xTM (cant see much, but at least get them in your sights) • Protection for your high dry (blue band, 400xTM) • Drop of oil • Oil immersion lens • ONLY USE FINE FOCUS ADJUSTMENT!!! • When done, use lens paper to clean up your lens and the stage From the Virtual Microbiology Classroom on ScienceProfOnline.com
When obtaining a bacterial sample from a tube or plate of media do so gently! The bacteria is growing as a thin film on top of the media! Don’t scrape so hard that you have pieces of agar in your sample! • If obtaining bacterial sample from slant tubes: • never pick up test tube by the cap. • do NOT set cap down on lab bench • flame neck of the test tube before & after • obtaining sample. Image: E. coli growing on TSY agar in slant tube and in Petri dish, T. Port From the Virtual Microbiology Classroom on ScienceProfOnline.com
Making a Bacterial Smear & Simple Stain 1. Draw circles with wax pencil 1. With a wax pencil, draw 3 dime-sized circles on a slide. 2. Flip slide over so wax is on the bottom. 3. Use DI water dropper to place a very small drop of water inside each circle. * sterilize inoculation loop in microincinerator and let cool * 4. Using inoculation loop, obtain a small sample of Staphylococcus epidermidis from the source plate or tube, as instructor has demonstrated. Swirl loop in circle of water on left. * IMPORTANT!!!: Very gently obtain sample of bacteria from the source plate or tube. The bacterial colonies are found growing on the surface of the TSY agar. DO NOT remove agar with your sample!! * sterilize inoculation loop in microincinerator and let cool * 5. Using inoculation loop, obtain a small sample of Escherichia coli (E. coli) from the source plate or tube. Swirl loop in circle of water in the middle. * sterilize inoculation loop in microincinerator and let cool * 6. Using inoculation loop, obtain a small sample of Bacillis subtilis from the source plate or tube. Swirl loop in circle of water on right. * sterilize inoculation loop in microincinerator and let cool * 7. Heat fix the slide on top of your microincinerator. Allow it to stay in the platform for 5 minutes after water has completely evaporated. This kills the bacteria and sticks it to the slide. 8. Stain the slide with crystal violet for 45 seconds then observe under oil immersion. 3. Put a drop of water in each circle. 4 - 6 Inoculate circle of water with each of the following: 1. Staph epi 2. E. coli, 3. Bacillus subtilis. 8. After heat fixing on microincinerator, stain with crystal violet, rinse, then look at with scope. From the Virtual Microbiology Classroom on ScienceProfOnline.com
Although you will be using 100xTM to get your specimen in focus, you will not be able to see individual bacteria until you use the 1000xTM oil immersion lens. Bacterial smear at 1000xTM (oil immersion) Bacterial smear at 100xTM FYI: The example above is Staph. Remember, you want to use the microscope with a built in camera so that you can take pictures of the bacteria you see at 1000xTM. From the Virtual Microbiology Classroom on ScienceProfOnline.com Images: Both images Staphylococcus, by T. Port
Confused? Here are links to resources that further explain microscopy: • Microscopy Laboratory Main Pageon the Virtual Microbiology Classroom of Science Prof Online. • Compound Microscope Parts and Usevideofrom Science ProfOnline. • How to Make a Wet Mount of an Elodea Plant Cellvideo from ScienceProfOnline. • How to Make a Wet Mount of an Onion Epithelial Cellvideo from ScienceProfOnline. • How to Make a Wet Mount of a Cheek Cellvideo from ScienceProfOnline. • Microscope Maniacrossword puzzle. • Viewing Bacteria Under Oil Immersion, SPO Lab Notes article. • How to Prepare a Microscope Slide of Bacteria, SPO Lab Notes article. • Prokaryotic Cellinteractive diagram from Cells Alive website. • “Germs”. Music by Weird Al Yankovic. Video by RevLucio. Smart Links (You must be in PPT slideshow view to click on links.) From the Virtual Microbiology Classroom on ScienceProfOnline.com
Are microbes intimidating you?Do yourself a favor. Use the…Virtual Microbiology Classroom (VMC) !The VMC is full of resources to help you succeed, including: practice test questions review questions study guides and learning objectives You can access the VMC by going to the Science Prof Online website www.ScienceProfOnline.com Images: E. coli, Giant Microbes; Prokaryotic cell, Mariana Ruiz