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Culture medium: nutrient source for growth/culturing, nutrient agar Label: initials, culture name, date Size, color,

shape, margin, opacity, elevation, surface, consistency, odor Microscope (compound light microscope) works via resolving power (ability to distinguish between 2 objects or see object details) and contrast (differences in color/brightness) Base, arm, ocular lens (eyepiece, 10x), rotating nosepiece, objective lenses (4x, 10x, 40x, 100x), stage, stage clips, stage knobs, coarse focus, fine focus, light intensity, condenser lens assembly (focuses light through slide), iris diaphragm (works like a shutter) Light wont turn on: light intensity knob, plugged in, partners scope Fungi spores, protists trophozoite/cyst, bacteria shape Shapes: spheres (coccus), rod-like (bacillus), spiral (spirilla/spirochetes) Arrangements: diplo, strepto, staphylo, coccobacilli Aseptic: sanitize/disinfect, hands, flame (loops, needles, tube mouths) before & after, dont set stuff down, open minimally Streak isolation: isolate bacterial colony (pure culture), 4 passes (top left), flame loop between streak Smear: bacterial cells mixed w/ water spread thinly/evenly, air dry, heat-fix (2-4x) Simple stain: discern shape/arrangement, meth blue/cv/safranin; morphology: bacilli, cocci, spirillum, spirochete, vibrio (bent), pleomorphic (no shape); arrangement: staphylo, strepto, diplo, tetrad Differential stain: differentiates cells, 2 or more stains Gram stain: diffentiate cells w/ thick peptidoglycan CW (gram +) vs LPS outer membrane (gram -) o Smear (dry, heat-fix) o CV (primary stain): stains pep CW (gram +) & outer membrane (gram -) o Iodine (mordant): increases affinity of primary stain to pep CW, also denatures proteins/kills cells o Decolorizer (95% eth alc): washes away outer membrane (gram -) rendering them colorless o Safranin (counterstain): stains colorless (gram -) pink Endospores: highly modified cellular structure that remains dormant until environment becomes favorable for growth; resistant to heat, drying, UV, freezing, disinfectants/antiseptics), gram + bacteria Endospore stain: differential, detects endospores, MG is water-soluble (doesnt adhere well) and vegetative cells disrupted by heat, therefore MG rinses easily, and takes up counterstain o Smear (dry, heat-fix 20x to soften spore coat, keratin) o Cut bibulous paper for slide o Malachite green (15-20 min) o Remove paper & decolorize o Safranin counterstain Minimal media: only ingredients essential to life of particular bacterium (not all bacteria can grow) Complex media: known components as well as ingredients w/ unknown make-up

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