1. About Science Prof OnlineScience 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/ Alicia Cepaitis, MS Chief Creative Nerd Science Prof Online Online Education Resources, LLC alicia@scienceprofonline.com From the Virtual Microbiology Classroom on ScienceProfOnline.comVirtual Microbiology Classroom ScienceProfOnline.com Image: Compound microscope objectives, T. Port Tami Port, MS Creator of Science Prof Online Chief Executive Nerd Science Prof Online Online Education Resources, LLC info@scienceprofonline.com

2. Laboratory Exercise 1b How to Use a Compound Microscope Viewing a Simple Stain of Bacteria Under Oil Immersion Aren’t They Cute? Image: Simple stain of Staphylococcus, T. PortStaphylococcus ** Please plug in your microincinerators ** From the Virtual Microbiology Classroom on ScienceProfOnline.comVirtual Microbiology ClassroomScienceProfOnline.com

3. 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.bacterial smear Q: What does the term parfocal mean? Image: Microscope objective lenses, T. Port From the Virtual Microbiology Classroom on ScienceProfOnline.comVirtual Microbiology ClassroomScienceProfOnline.com

4. 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! Image: Microscope objective lenses, T. Port From the Virtual Microbiology Classroom on ScienceProfOnline.comVirtual Microbiology ClassroomScienceProfOnline.com

5. 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.bacterial smear Put a drop of immersion oil directly on each of the three bacterial smears on your slide, then switch to the oil immersion lens. Image: Microscope objective lenses, T. Port From the Virtual Microbiology Classroom on ScienceProfOnline.comVirtual Microbiology ClassroomScienceProfOnline.com

6. Observing bacteria under oil immersionObserving 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.comVirtual Microbiology ClassroomScienceProfOnline.com

7. 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! From the Virtual Microbiology Classroom on ScienceProfOnline.comVirtual Microbiology ClassroomScienceProfOnline.com Image: E. coli growing on TSY agar in slant tube and in Petri dish, T. Port E. col 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.

8. Making a Bacterial Smear & Simple Stain 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.inoculation loop * 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.microincinerator 8. Stain the slide with crystal violet for 45 seconds then observe under oil immersion.oil immersion Draw circles with wax pencil Put a drop of water in each circle. Inoculate circle of water with each of the following: 1. Staph epi 2. E. coli, 3. Bacillus subtilis. After heat fixing on microincinerator, stain with crystal violet, rinse, then look at with scope. 1. 3. 4 - 6 8. From the Virtual Microbiology Classroom on ScienceProfOnline.comVirtual Microbiology ClassroomScienceProfOnline.com

9. Although you will be using 100 xTM to get your specimen in focus, you will not be able to see individual bacteria until you use the 1000 xTM oil immersion lens. Bacterial smear at 100xTM Bacterial smear at 1000xTM (oil immersio n) FYI: The example above is Staph. Images: Both images Staphylococcus, by T. PortStaphylococcus 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.comVirtual Microbiology ClassroomScienceProfOnline.com

10. Confused? Here are links to resources that further explain microscopy: Microscopy Laboratory Main Page on the Virtual Microbiology Classroom of Science Prof Online. Science Prof Online Compound Microscope Parts and Use video from Science ProfOnline. Compound Microscope Parts and Use How to Make a Wet Mount of an Elodea Plant Cell video from ScienceProfOnline. How to Make a Wet Mount of an Elodea Plant Cell How to Make a Wet Mount of an Onion Epithelial Cell video from ScienceProfOnline. How to Make a Wet Mount of an Onion Epithelial Cell How to Make a Wet Mount of a Cheek Cell video from ScienceProfOnline. How to Make a Wet Mount of a Cheek Cell Microscope Mania crossword puzzle. Microscope Mania Viewing Bacteria Under Oil Immersion, SPO Lab Notes article. Viewing Bacteria Under Oil Immersion How to Prepare a Microscope Slide of Bacteria, SPO Lab Notes article. How to Prepare a Microscope Slide of Bacteria Prokaryotic Cell interactive diagram from Cells Alive website. Prokaryotic CellCells Alive “Germs”. Music by Weird Al Yankovic. Video by RevLucio.Germs (You must be in PPT slideshow view to click on links.) From the Virtual Microbiology Classroom on ScienceProfOnline.comVirtual Microbiology ClassroomScienceProfOnline.com

11. 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:VMC 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 www.ScienceProfOnline.com Images: E. coli, Giant Microbes; Prokaryotic cell, Mariana RuizE. coliProkaryotic cell