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205 Cards in this Set
- Front
- Back
- 3rd side (hint)
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Purpose of aseptic technique |
Insures no contamination of microorganisms into work place, cultures you are working with, or to others present. |
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Loops and needles need to be .. |
Sterilized until they are red hot |
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Why do we flame tools after using them? |
To kill left over organisms |
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TSA |
Tryptic Soy Agar |
Tryptic soy |
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TSB |
Tryptic Soy Broth |
Tryptic soy |
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Label all sterile media with.. |
Your name, media type, organisms used, dateand lab hour. Label on bottom of lid |
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Smear prep is what is determined? |
Prepares org for staining. Used to determine 1. cell shape, 2. Cell arragemnet and 3. internal structures. |
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Most stains we do utilize ? |
Smear prep to place orgs on slide |
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The key to successful smear.. |
Make sure smear is not too thick, too many cells make it hard to see shape or arrangement of cells |
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Organisms in solid media clump together and can be dispersed how? |
By adding a few loopfuls of water |
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When taking from a solid media do u take a lot or a lil dab? |
Less is better. U take a small dab |
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A solid media may need to be mixed with a little ______ on smear prep |
Deionized Water |
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This experiment ties in with many other experiments |
Smear prep |
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What is the purpose of heat fixing the slides? |
Kills organisms and adheres them to the slide so they r not washed off during staining |
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Simple staining |
Makes it easier to visualize bacterial cells on slide by using dyes |
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Simple staining has 2 types of dyes |
1. Basic dye (+)charged chromophore -Works Well with bacteria cells -Stains inside of cell 2. Acidic dye (-)charged chromophore -REPELLED by bacteria -stains outside of cell |
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Bacteria has what charge ? |
Bacteria is negatively charged |
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Simple stains are used to determine? |
Bacterial cell morphology (cell shape) |
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3 common bacteria shapes |
Bacilli (rods) Cocci (spherical) Spiral or curved |
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Dye used in simple stain |
Methylene blue |
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Negative staining |
Determines size, and shape of bacteria |
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Neg stains are ___? |
Acidic dyes & r repelled by bacteria |
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Negative stains stain___? |
The background of cell. The bacteria itself will appear transparent against dark background. |
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India ink is (-) and is used in |
Negative staining |
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Acidic dye is? |
Neg charged chromophore |
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Chromophore is? |
A molecule that gives dye its’ color. |
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India ink is used in which experiment? |
Negative stain |
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In a negative stain why does it stain outside the cell? |
Because of their charge |
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Capsule stain is what kind of stain and dye? |
It is a differential stain. It uses acidic and basic dye |
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2 distinct Layer tht surround some bacterial cells |
1.Capsule 2. Slime or biofilm |
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Capsule is? |
-Distinct gelatinous layer -Attached tightly to bacteria -Most capsules are made of polysaccharides aka glycocalyx |
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Slime or biofilm |
-A diffuse n irregular layer of extracellular matrix -Loosely attached to bacteria, and can be washes off |
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Do we heat fix capsule stain? |
No, it will melt capsule n we wont b able to dye or see ut |
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What function do capsules have? |
They protect the bacteria from being engulfed or destroyed. Also helps them attach to solid surfaces |
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How does capsule stain differ from other stains? |
-We use negative staining for the first part -there's no heat fixing -all the sides are air-dried |
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What would heat fixing a capsule stain do? |
It would destroy her shrink the capsule we will be unable to view it |
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What is the purpose of gram staining? |
To provide identification of the unknown bacteria |
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Gram staining is an example of what staining technique? And what does that technique mean? |
Differential stain: to tell the difference between cell or cell structures due to dissimilar staining |
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What is the cell wall feature in a gram positive cell? |
90% peptidoglycan 10% techoic acid |
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What is the cell wall feature in a gram positive cell? |
90% peptidoglycan 10% techoic acid |
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What is the cell wall feature for a gram negative cell? |
90% outer membrane 10% peptiodoglycan |
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Gram positive will show what color and why? |
Purple because of the thicker peptidoglycan layer( cell wall) |
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What color will the gram negative be and why? |
Pink/red because of the thinner peptidoglycan layer(cell wall) |
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If the reagent is just heat fixed, what will gram positive and gram negative look like? |
Clear |
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If the reagent is crystal violet what color would gram positive and gram negative be? And what is crystal violet? |
Purple and crystal violet is a primary stain. |
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The reagent is Gram’s Iodine, what is it’s purpose and what color will gram positive and gram negative be? |
It is a mordant and both colors are purple. |
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What will 95% Ethanol do to gram positive and gram negative? And what is the purpose of ethanol? |
Gram positive stays purple and gram negative is clear. Ethanol is a decolorizing agent. |
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What is the reaction of Safranin towards gram positive and gram negative? And what is the Safranin’s purpose? |
Gram positive is purple and gram negative is pink/ red. Safranin is a counterstain. |
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Gram positive has a ______ cell wall. |
Thick |
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Gram negative has a _____ cell Wall. |
Thin |
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Under oil immersion capsule stains appear as |
Halos around pink/red cell w a dark background |
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Acidic dyes are |
Negatively charged |
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Dyes used on capsule stains |
Congo red - acidic-stains background maneval- basic dye- stains the inside of cell |
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Basic dyes |
R positively charged n stain the inside of cell |
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On what experiment does the cell appear to have a halo under oil emmersion? |
Capsule staining |
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Capsule staining uses what dye? |
Congo red or nigrosin |
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Capsule staining uses what dye? |
Congo red and manevals |
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What are the bacteria that belong in rod (bacilli)? And what shape? |
Bacilli is flat, round or tapered at the end. 1. Coccibacillus 2. Bacillus 3. Fusiform |
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What are the bacteria that belongs in cocci (spherical)? |
1. Steptococci (chain) 2. Staphylococcus (cluster) 3. Diplococci (two cocci) 4. Tetrads (4) |
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What are the bacteria that belongs is spiral or curved? |
1. Spirochates 2. Comma 3. Spirillum |
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What are the bacteria that belongs in spiral or curved? |
1. Spirochates 2. Comma 3. Spirillum |
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What are the dyes used in capsule staining? And what are their dyes? |
Mordant: Congo red- acidic dye Counterstain: Manevels-basic dye |
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The total charge of a cell is posifive |
False |
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Total magnification of high power |
400 x |
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The shaettr futln method is an alternative for what lab producer? |
Spore stain |
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Congo red |
Acidic |
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Organisms picked up thru out the day are |
Transient microrganisms |
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Dye used in simple stain |
Methadine blue |
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S aureus,effacilies, and b. Serious |
Are all gram + |
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Positive result for acid fast stain |
Pink |
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Positive spore stain has |
Green spores |
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Who developed gram staining |
Hans Christian gram |
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Staning technique tht differentiates bacterial cells from eukaryotic cells |
Gram stain |
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Wich of the following was not a tropical used in the antiseptics test |
95% ethinal alcohol |
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Dye used in acid fast stain to dye cells with myolic acid membraine |
Carbo fusion |
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After alcohol fluds the slide in a gram stain, gram + cells remain purple and the gram -cells are |
Colorless |
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Safranin is a dye used in |
Gram stain and spore stain |
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What dyes cells in a capsule stain |
Menevals |
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The mordant in acid fast stain is |
Heat |
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India ink is a dye used in |
Neg stain |
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A high % transmission value indicates |
Low number of organisms |
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The organism used in the motility test was |
P.vulgaris |
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Purpose of this procedure is to decrease the concentration of cells by a known volume |
Serial dilution |
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Acid fast cells stain in the acid fast stain |
Pink |
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The total charge of cell is neg |
True |
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Optical |
What i can see w my eyes |
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Optimal |
Best living conditions |
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What is the purpose of oil immersion |
The oil replaces the air space. It prevents refraction at a glass air interface. Meaning allows less light to go thru so we see org better |
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What r 2 possible errors which could have occurred if u find ur subcultures had no growth after being incubated? |
1. Forgetting to inoculate 2. Could have killed bacteria w heat from un-cooled needle |
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Four microscopic morphologies of bacteria |
Cocci, coccobacilli, bacilli, and spirals |
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What is the real name of the white rimmed objective |
Oil emmersion |
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Why is sheeps blood used as media? |
Fastidious bacteria Requires xtra nutrients |
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What is the name for the blue rimmed objective? |
High dry |
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What is the function of the iris diagphram? |
Adjustment for light thru the abee condenser , want it in the middle, light should not be blinding |
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5 characteristics of bacterial colonial morphologies? |
1. Size 2. Shape 3. Color 4.texture 5. Density |
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Which stain makes the cell look pink |
Acid fast stain |
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What two things shud u always do to enhance isolation of bacteria |
1.Flame and cool needle 2. Firm dab , dont scoop |
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What r the functions of the abbe condenser? |
1 collect light 2 condense light 3 alternate light source |
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Why are culture tunes held at an angle? |
To prevent air contamination |
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4 places normal flira exists |
1. Gi tract 2 vagina 3 throat/mouth 4sinuses |
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Purpose of inverting plates during incubation? |
To prevent buildup of condensation that would ruin the culture |
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What and where is agar from |
Solid form media Comes from seaweed |
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What is total magnification for low dry |
100x |
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What is streaking for isolation |
To break up colonies from 2 species |
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What group of organisms are considered acid fast? |
Mycobacterium |
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Name of yellow rimmed objective |
Low dry |
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Color of serratia at room temoerature |
Red |
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What 2 parts of microscope r used to carry or support the microscope? |
Arm and base |
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Total magnification of scannibg onjective |
40x |
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Total magnification for high dry |
400x |
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Red color objective is called |
Scanning |
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What is media |
Broth or agar used Base" for bacteria to be added to |
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Parfocal lensesallow for us to b able to go between objectives w little adjustment of ... |
Fine focus |
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What is normal flora |
Grows on different mucosal surfaces of the body |
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Total magnification of oil immersion |
1000x |
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3 optical parts to microscope? |
Eye piece, abbe condenser, objective lense |
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Equation for total magnification |
Objective magnification x ocular magnification |
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Y shud a petri dish not be left open for an extended period of time? |
Air cobtamination |
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What r ways to tell media is sterile |
Run controls No floaters in agar No growth in broth |
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Gram staining is an important way to identify... |
Unknown bacteria |
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Gram stain |
Is A differential stain |
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Gram + and gram - are different based on their |
Cell wall , composition , and structure |
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Cell wall features in gram + cell is |
90% peptidoglycan pkus 10% teichoi acids |
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Cell wall features in gram - is |
10%peptidoglycan plus 90% outer membrane |
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Gram + cells will appear |
Purple |
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Gram - cellz will appear |
Pink/red |
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Why do gram + stain purple but not gram - |
Because gram + has a much thicker peptidoglycan layer |
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Mordant |
Glue or sticky stuff in middle such as cement between brinks |
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Acid fast stainibg is used to |
Identify mycobacterium and nocardia species of bacteria |
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Mycobacterium and nocardia have a |
Cell wall lipids w waxy materials |
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What is the waxy layer on cells called? |
Mycolic acid |
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This layer makes certain stainingvprocedures ineffective? |
Mycolic acid |
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If it has mycolic acid |
Its red |
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If it stains blue there is no |
Mycolic acid |
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In acid fast stainubg carbolfusion stain red and methtlene blue |
Stain blue |
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When mycolic acid is precent on bacteria |
Certain stains r i effective |
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If bacteria cell is acid fast |
Will stain red |
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On acid fast non-acid will stain |
Blue |
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What kind of staon is an acid fast stain? |
A differential stain |
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Why do cells w mycolic acid stay red? |
Bc when decolorized w alcohol, it is ineffective bc the acid fast + bacteria are resistant to alcohol. |
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Mycolic acid |
Protects bacteria cells |
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We heat fix carbofission on acid fast stain |
To allow pink dye to penetrate waxy cell wall |
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Why do we test acid fast n non acid fast together |
To differentiate which has a waxy layer |
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Endospores stain |
Green |
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Do cells sexually reproduce while in endospore state? |
No |
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Certain gram + species for |
Endospores when they are stressed |
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When r endospores formed ? |
When gram + cells are stressed |
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What makes cells stressed |
When they r faced with unfavorable conditions like low or high ph, temo too low or high, if ainething is trying to eat them, or if little nutrients are availablr |
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Endospores are utilized as |
Resting stages tht allow bacteria to survive unfavorable conditiins |
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Once environmental conditions become favorable endospores can go through germination process and form |
Vegetative cells |
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Vegetative means |
Healtjy, able to reproduce , eat |
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Resistant to heat, radiation, acids, and chemicals |
Endospores |
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Endospores ate resistant to heat ect because they r made of |
Protein coat |
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To destryo the hard protein coat of an endospore with heat we must |
Steam at 121*c for 15 to 20 mins by heat fixing |
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Endospores are not easily penetrated by stainibg procedures |
True |
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What needs to b applied to help stain enter endospospore? |
Heat |
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In endospore stain the vegetative (happy) cells will appear red due to seffarin and the endospores cells |
Green fue to malachi green dye |
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What is a differential stain |
Distinguishes between 2 diff organisms |
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Turbidity |
Cloudy |
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Turbidity indicates the bacteria |
Can move, is motile |
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Motility most often found in |
Spirochetes and rod shaped bacteria |
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What is the major orgabelle used for motility |
Flagella |
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Flagella are used primarily to |
Mive away from harm or move toward nutriengs |
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Why do we stain flagella |
Bc they r so thin they r hard to see on their own |
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Motility can b observed by |
Semisolid media, wet mount, or hanging droo |
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When looking at organisms on slide and u see movemement this is called |
Not true motility , brownian motion |
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Brownina motion is when cella are under glass and they appear to move |
Cells are only vibrating in straight line in 1 dirrection, this is non tru mobility |
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Dye in semi solid motility ex |
Allows to see where bacteria moved thru |
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Why do cells mive |
To run away from danger or get closer to trients |
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Which org will exhibit tru motility |
P. Vulgaris |
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Main point of pour plate technniqu |
Diluting colonies down to isolate a specific organism |
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2 common pure culture techniques are |
pour plate and streak plate |
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Pure culture |
Helos get a single organism from a mixed culture |
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B bacillis form endospores when in harsh conditions and when uv light is remived |
They return to vegetative (happy) state |
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What is the term antimicrobial mean? |
An agent that kills it inhibits antimicrobials |
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What does the term antibiotics mean? |
Low molecular weight antimicrobials produced from microorganisms that kill or inhibit other bacteria. |
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What method has the term antibiotics and antimicrobials? |
The Kirby bauer method |
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Antiseptic is? |
A substance that kills or inhibits microbial growth and gentle enough to apply to living tissue |
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Disinfectant is? |
A substance that is applied to inanimate objects that kill or reduce microorganisms but harsh on living tissue |
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pH and microbial growth have three groups what are they? |
Neutrophiles, acidophiles, alkaphiles |
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pH and microbial growth have three groups what are they? |
Neutrophiles, acidophiles, alkaphiles |
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What are the primary lethal effects of uv light? |
Mutagenic properties |
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pH and microbial growth have three groups what are they? |
Neutrophiles, acidophiles, alkaphiles |
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What are the primary lethal effects of uv light? |
Mutagenic properties |
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What light is most germicidal? |
260 nm UV because dna absorbs the uv light and bonds are broken at its maximum |
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Pure culture |
Obtain single organism from a mixed culture |
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Two methods of pure culture |
Pour plate and streak plate |
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Motility is often found in |
Spirochetes and rod bacilli |
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Purpose of flagella |
They move away from harmful substances and move towards nutrients. They are thin structures so they need to be stained to be viewed easier |
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What are they methods of motility? |
Hanging drop, wet mount and semi solid media |
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Acid fast staining |
Used to identify mycobacterium and Nocardia. These both have mycolic acid |
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Acid fast staining is what stain |
A differential stain |
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Acid fast staining uses what dye |
Carbolfuschin |
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Nom acid fast staining uses what dye |
Methylene blue |
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INITIATION begins at a specific nucleotide sequence called |
The origin of replication (Ori) |
Lec 3 |
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Topoisomerase ii (aka GYRASE) |
Uncoils the DNA |
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_________ then separates the DNA strands by breakin the hydrogen bonds between nitrogenous base pairs. |
Helicase |
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What occurs in the initiation step ? |
Topoisomerase ii Helicase |
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Steps in Elongation (of dna replication) |
1)Dna helicase opens repkication fork 2 ) On leading strand Polymerase iii followshelicase and adds on nucleotides according to 3' n 5' THIS IS CONTINUOSLY |
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More elingation steps |
3) on laggin strand dna polymerase iii replicates Okazaki fragments (DISCONTINUOUS) 4) DNA ligase connects Okazaki fragments together for copy of lagging strand |
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Ligase means |
To tie together |
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What happens at the end of elongation step in dna replication? |
U end up w 2 interlocking circles of dna [NOW THEY NEED TO B SEPERATED] |
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Termination steps |
Bacterial topoisomerase iv Which is responsible for separating the chromosomes and making 2 individual ones |
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What enzyme is required for DNA synthesis in bacteria? |
DNA polymerase iii |
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Wht does pol iii do? |
It adds individual nucleotides according to their COMPLIMENTARY BASE a + t and c+g |
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