Bio 2EE3

Lab 1 pre-lab talk

Identifying Mi b by Id tif i Microbes b Microscopy and Physiology

Laboratory Safety y y
You must have Biosafety Level 1 Training before working in the laboratory: register in and complete HTH SCI 1BS0
Lab coats: Coats must be worn to protect you and your clothing from stains and corrosive reagents. Biohazards: T t all organisms used as th Bi h d Treat ll i d though th are d h they dangerous, and b careful d be f l to avoid any direct contact with them. Do not drink/eat in the laboratory and ASK your TAs in case you should have any questions. Plates not used any more should be disposed of in the Biohazard waste cartons in the lab. Spills/Sterilization: There are bottles of a disinfecting solution (5% Dettol) at each bench for wiping your work areas before the beginning and after the end of work. If a culture is spilled, disinfect the area with 5% Dettol. Wash your hands with the bacteriocidal soap provided before leaving the laboratory laboratory. Glass waste: Dispose of slides or damaged glassware that has come in contact with microorganisms in the Biohazard sharps containers. Dispose other glass waste in the Glass waste cartons cartons.
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Bio 2EE3

Lab 1 pre-lab talk

Relative sizes of different cells.

3 Slonczewski and Foster: Figure 2-4

Bio 2EE3

Lab 1 pre-lab talk

Common shapes of bacteria.

4 Slonczewski and Foster: Figure 2-6

Bio 2EE3

Lab 1 pre-lab talk

Microscopy and X-ray crystallography, range of resolution. f l ti

Slonczewski and Foster: Figure 2-7

Bio 2EE3

Lab 1 pre-lab talk

Anatomy of a compound microscope.

Slonczewski and Foster: Figure 2-17

Bio 2EE3

Lab 1 pre-lab talk

Use of immersion oil in microscopy.

Slonczewski and Foster: Figure 2-16

Bio 2EE3

Lab 1 pre-lab talk

Lab 1 part 1 Work in groups of 4 Gram s Grams stain on each culture: observe morphology and Gram result Malachite Green stain on all cultures: observe presence or absence of endospores
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Bio 2EE3

Lab 1 pre-lab talk

Gram staining of bacteria (a type of differential stain). stain)

Sample from the mouth, showing Gram positive and Gram negative bacteria.

Slonczewski and Foster: Figure 2-21

Bio 2EE3

Lab 1 pre-lab talk

The Grams Stain a differential stain

Slonczewski and Foster: Figure 2-22

Slonczewski and Foster: Figure 2-23

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Bio 2EE3

Lab 1 pre-lab talk

Lab 1 part 2 Work in groups of 4 Inoculate all cultures into thioglycollate agar Inoculate all cultures into phenol red broth Catalase test on culture agar slants
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Bio 2EE3

Lab 1 pre-lab talk

Oxygen-related growth zones in a standing test tube.

Thioglycollate agar Inoculate with a needle to the bottom of the tube. Observe where Ob h growth occurs Indicates oxygen requirements of that organism
Slonczewski and Foster: Figure 5-19 12

Bio 2EE3

Lab 1 pre-lab talk

Clinical tests based on fermentation.

Phenol red broth pH indicator Different fermentation pathways result in acid and/or gas production Gas collects in the Durham tube

Slonczewski and Foster: Figure 13-23

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Bio 2EE3

Lab 1 pre-lab talk

Generation and destruction of reactive oxygen species (ROS). (ROS)

Slonczewski and Foster: Figure 5-20

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Bio 2EE3

Lab 1 pre-lab talk

Lab 1 part 3: to do individually Plate an environmental sample to obtain single colonies: Soil, water Sample from your teeth Yogurt Fingers Cultures A E (before the catalase test!) NOTHING PATHOGENIC!
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Bio 2EE3

Lab 1 pre-lab talk

Dilution streaking technique.

Slonczewski and Foster: Figure 4-12

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Bio 2EE3

Lab 1 pre-lab talk

Assessment Complete the informal lab report Share results but the lab report must be done individually Due ONE week after the lab dropboxes Morning labs by 12 noon Afternoon labs by 4pm
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