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Streak plate for Environmental Sampling
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4 Quadrant Streak Plate 1. swab 2. loop 3. loop 4. loop
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Four Quadrant Streak Plate
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Red algae site:oceanexplorer.noaa.gov, used with permission
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Agar Properties Melts 100 C but liquid till 40 C Not toxic Not “degraded” Lacks nutrients
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Proper Labeling Name Date Name microbe/source of inoculum Ex # Miscellaneous info
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AsepticAseptic Technique vs. Universal Precautions Why??? How???? When???? Microbelibrary ASM, used with permission
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To Do Take 2 TSA plates and two broths Properly label them Wet swab with sterile water…sample body with 1 swab….streak quadrant 1 of TSA plate Take swab and place in TSB tube Repeat for “environment” plate.
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4 Quadrant Streak Plate 1. swab 2. loop 3. loop 4. loop
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spores thallus Fungal Classification: molds
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Fungal classification: yeasts
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Budding KM
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Dimorphism
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Penicillium notatum
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Penicillium 100x 400x KM
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Penicillium spores
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Rhizopus 100x KM
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Rhizopus
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Aspergillus 100x 400x KM
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Candidiasis Cutaneous fungal infections
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Tinea pedis Dermis.net, used with permission
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Tinea corporis Dermis.net, used with permission
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Tinea cruris, ringworm of the groin Cutaneous fungal infections
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Candidiasis www.meddean.luc.edu
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Sporotrichosis DermAtlas, used with permission
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Subcutaneous fungal infections Sporotrichosis CDC Dr. Lucille K. Georg, used with permission
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A few last points Systemic mycoses True Fungal infections Opportunistic infections
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The life cycle of Aspergillus Spores inhaled Germination Mass of hyphae (plateau phase) Hyphal elongation and branching http://www.aspergillus.org.uk/index.html & Fungal Research Trust, used with permission
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Aspergillosis in the brain WebPath, used with permission
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Mucomycosis eye infection CDC, used with permission
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To Do Part II Prepare Fungal Cultures Glass slide agar Fungal smear
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Staining
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Principles of Staining Stain or dye: A molecule that can bind to a cellular structure and give it color (contrast) 1. Positively charged or Basic Dyes 2. Negatively charged or Acidic Dyes Simple stain: makes use of a single dye and reveals basic cell shapes and arrangements Differential stain: makes use of two or more dyes and distinguishes between organisms based on structural differences Wiley
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Mechanism of Direct Stains Add a positively charged blue dye
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Reasons for Heat Fixation Kills the Cells Adherence Coagulation Prevents Autolysis
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Procedures SE BMEC Right NOW: Make 2 identical slides. Label 1 Gm and the other Dt 1.Smear (both slides) 2.Air dry (both slides) 3.Heat fix (both slides) Gm KM
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Mechanism of Negative stainingNegative Field of view Microbe Add a negatively charged blue dye Animation courtesy of ASM MicrobeLibrary, used with permission
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Watch Demo on how to make a smear for a indirect stain
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Examining Indirect Stains KM
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Example of a Negative Stain KM
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1-2 seconds Gram StainGram Stain ASM MicrobeLibrary, used with permission
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What do you observe? What do you conclude?
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Always look carefully Used with permission
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Always look carefully KM
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Bacterial Shapes KM
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Bacterial Arrangements KM
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Gram reactions KM Used with permission
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Staining Bacteria
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1-2 seconds Gram StainGram Stain ASM MicrobeLibrary, used with permission
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