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M200 Lab Midterm
| Question | Answer |
|---|---|
| compound microscope | using series of lenses and a source of bright light, it magnifies and illuminates minute objects--invisible to the naked eye |
| eyepiece | what you look into; magnification of 10x |
| nosepiece | holds objective lenses |
| scanning power | magnification is 4x total magnification is 40x always start with to begin focusing |
| low power | magnification is 10x total magnification is 100x |
| high power | maginification is 40x total magnification is 400x |
| oil immersion | magnification is 100x total magnification is 1000x oil prevents light from scattering and improves resolution (refractive index) |
| mechanical stage | holds the specimen |
| coarse focus knob | focuses by raising the stage initial focusing always done with the scanning lens |
| fine focus knob | fine tuning focus doesn't move any parts of the microscope |
| adjustment knobs | moves specimen big-up and down little-left and right |
| substage condenser | focus light on specimen |
| iris diaphragm | adjusts the light |
| base | serves as a support; holds the illuminator |
| parfocal | once in focus...it should remain in focus |
| resolution | the ability to distiguish between two points |
| inversion | upside down and backwards demonstrated by the e slide |
| focal length | length between the lens and the stage |
| field of view | as magnification increases, the field of view decreases magnification and field of view are inversely related |
| total magnification | magnification of the objective lens times the magnification of the eyepiece |
| aniline dyes | synthetic organic dyes made from coal-tar products allows the shape of bacterial bodies to be seen react in either an acidic, basic, or neutral |
| acidic dyes | have anions that combine with the base in stained cell to form salt |
| basic dyes | have cations that combine with the acid in stained cell to form salt bacterial cells rich in ribonucleic acids stain well |
| neutral dyes | combination of acid and basic used to stain more complex cells |
| simple stain | allows the identification of shape and arrangement of the stained bacteria this staining doesn't identify between specific species doesn't kill the bacteria |
| bacillus | rod-shaped |
| coccus | sphere; round can take a football shape |
| spirillum | spiral (has at least 2 or 3 curves) |
| Brownian movement | false movement that can be caused by vibration of the table or microscope |
| arrangements of bacillus | single bacilli diplobacillus streptobacillus palisade (unique to bacillus) |
| arrangements of coccus | diplococcus (rarely found singly) tetrads (unique to coccus) streptococcus staphylococcus |
| arrangements of spirillum | no arrangements taken spirilla are short spirochetes are long tightly coiled |
| gram stain | stains bacteria different colors depending on it being gram pos/neg makes it possible to see bacteria to see bacteria clearly, but doesn't distinguish between organisms of similar morphology |
| gram positive | stained by the crystal violet (primary stain) purple |
| gram negative | loses the first dye when washed with the decolorizer; takes color of counterstain (safranin) red |
| gram positive cell wall | large peptidoglycan layer that allows resistance of the decolorization stage the thick, tightly packed peptidoglycan in which crystal violet-iodine complex becomes trapped, thereby enabling cell to resist decolorization |
| gram negative cell wall | small peptidoglycan layer and large NPS that undergoes decolorization well have a high concentration of lipids in their outer membranes that are dissolved in decolorizer and are washed away along with the crystal violet-iodine complex |
| procedure for gram stain | 1. sterilize loop 2. 1 or 2 drops of water 3. inoculate on slide 4. let slide air dry 5. heat fixation 4x4 method 6. apply stains |
| primary stain | crystal violet leave on for 1 min dyes both cell walls |
| mordant | IKI- Gram's iodine leave on for 1 min; then rinse and blot dry combines with crystal violet to form an insoluble salt forms a complex in gram pos cell wall |
| decolorizer | ethyl alcohol leave on for 15 secs; then rinse and blot dry crystals remain in gram pos cell wall( purple color remains; the outer membrane of gram neg are weakened and lipids are dissolved |
| counterstain | safranin leave for 1 min; then rinse and blot dry stains gram negative bacteria red |
| examples of gram positve bacteria | *all coccus are gram positive Staphylococcus aures Staphylococcus epidermis Micrococcus luteus Bacillus subtillus |
| examples of gram negative bacteria | Escheria coli (football shape) Enterobacteria Salmonella |
| Acid-fast stain procedure | 1. aseptically transfer bacteria and heat fixate 2. Place slide over boiling water w/ carbofuschin(primary stain) for 5 min 3. let cool and rinse 4. decolorize w/ acid alcohol about 15 gtts and rinse 5. Counterstain-methylene blue x 1 min; rinse |
| Ziehl-Neelson technique | using boiling water to heat stain; answer to a stand and burner set up |
| effect of acid fast staning on bacteria | acid fast positive is red; stained by carbofuschin acid fast negative is blue; stained by methyl blue |
| Mycobacterium | contain large amounts of lipids within thier cell walls--known as mycolic acids (resist staining) when stained wit basic dye and applied with heat the stain penetrates the lipid cell wall and reach the cell cytoplasm acid-fast pos resist decolorizer |
| Nocardia | partially acid fast resist decolrizer with weak sulfuric acid, but lose dye when treated with acid alcohol |
| spore stain procedure | 1. aseptically transfer bacteria and heat fixate 2. Place slide over boiling water w/ malachite green(primary stain) for 5 min 3. let cool and rinse 4. decolorize w/ acid alcohol about 15 gtts and rinse 5. Counterstain-safranin x 1 min; rinse |
| effect of spore stain on bacteria | spores will stain green ex Bacillus subtillus not spores will stain pink/red characteristic of Bacillus and Clostridium |
| negative stain | highlights the capsul and stains the background ex Proteus vulgaris 1. place 1 gtt of nigrosin on end of slide 2. add bacteria to nigrosin 3. using a 2nd slide evenly spread mixture on slide 4. air dry and heat fixate---4 times only (1x4) |
| flagella | protein structure not present in all bacteria its arrangement is a form of identification |
| monotrichous | single polar at one end |
| lophotrichous | mono- or bipolar collections |
| peritrichous | multiple flagella surrounding cell |
| amphitrichous | have flagella at each end |
| basal media | protein, carbohydrates, mineral salts allows growth of everthing that doesn't require special nutrients |
| fastidious | require special nutrients to grow ex. blood agar |
| facultative anaerobe | can tolerate presence of oxygen; all over test tube |
| obligate aerobic | needs oxygen; at surface only |
| obligate anaerobe | cannot tolerate presence of oxygen; at very bottom of tube |
| microaerophilic | needs just a little oxygen; just below the surface |
| turbidity | cloudiness of broth |
| pure culture | single, separated colony that will grow only one type of bacteria |
| mixed culture | several different species growing together; different color or different shape |
| colonial morphology | provide clues as to the identity of an organism; color, density, consistency, surface texture, shape, and size |
| Quadrant tech | aseptic tech purpose is to obtain pure isolated colonies |
| heat fixation | kills bacteria while still maintaining shape |
| MRSA | methillin resistant Staphylcoccus aureas common nosocomial infection |
| A B D C | A:compromised host-patient B:opportunistic bacteria C:chain of transmission D:nosocomial infection combine A, B, and C and you get a nosocomial infection |
| ubiquotous bacteria | can be found everywhere |
| autoclave | steam-pressure sterilizer 121 C for 15 min for 15 psi (pounds per square inch) |
| subculture | obtaining a pure culture from selecting a single colony from a mixture |
| differential media | contains special components that can be used by some microorganisms but not by others provides a color change to colonies or media |
| selective media | contains special components that suppress the growth of some microorganisms without seriously affecting the ability of others to grow |
| MacConkey's Agar (MAC) (light/pale) | Sel: gram neg Inhibitor:crystal violet (inhibits gram +) Diff:Lactose fermentation Indicator: neutral red (bacteria color change) lactose (+) is purple colonies; lactose (-) is opaque/colorless colonies |
| Mannitol Salt Agar (MSA) (hot pink) | Sel:gram pos (Staphylococcus & Micrococcus) Inhibitor:7.5% salt (inhibits gram -) Diff:mannitol fermentation Indicator: phenol red; mannitol (+) will turn media yellow; mannitol (-) media will remain hot pink |
| Eosin Methylene Blue (EMB) (dark red) | Sel:gram neg Inhib:eosin Y (inhibits gram pos) Diff:lactose fermentation Indicator:eosin Y and methylene blue; lactose(+)blue/black or brown colonies; lactose (-)opaque/colorless colonies E. coli is lactose (+) and gives a green metallic green sheen |
Created by:
marosado10
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