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Practical 1
| What is the recommended range for P1000 micropipette | 1000-100 |
| What is the recommended range for P100 micropipette | 100-10 |
| What is the recommended range for P10 micropipette | 10-0 |
| What do you call the part of the micropipette that allows you to view the volume? | volume display |
| What do you call the part of the micropipette that allows you to view the recommended volume? | |
| What do you call the part of the micropipette that allows you to adjust the volume? | volume adjuster |
| What do you call the part of the micropipette that holds the tips? | disposable tip attachment point |
| What is magnification? | The enlargement of an object |
| Objective magnification | magnification of the objective lenses 4x, 10x, 40x, 100x |
| Total magnification | magnification from ocular lenses 10x times the magnification of the objective lenses. |
| Field of view | Total surface area that is visible through the lens, as magnification increases FOV decreases |
| parfocal | focus is maintained when switching objective |
| paracentral | some centeredness is maintained when switching objectives |
| re-centering | adjusting centeredness after switching objectives |
| re-focusing | adjusting focus after switching objectives |
| compound light microscopes are both what | parcentral and parfocal |
| When should the coarse focus knob be used? | 4X only |
| what do you do at 100X | add immersion oil |
| Resolving power (resolution) | the amount of detail an image has, or the ability to distinguish between two adjacent objects as discrete entities |
| How do you increase resolution or resolving power? | by adding immersion oil, cleaning the objective lens, and using shorter wv light. |
| What does immersion oil do | refracts light and acts as a funnel directing light into objective lens |
| Contrast | How well an object stands against its surroundings/background |
| How do you increase contrast? | By adjusting light and adding stain |
| Depth of focus | thickness of a sample that appears in focus at a given magnification, as magnification increases depth of focus decreases |
| Working distance | the distance between the slide and the objective lens when the object is in focus, as magnification increases working distance decreases |
| division morphology | arrangement of cells after dividing |
| cell morphology | shape of individual cells |
| What does staining accomplish | adds contrast and differentiates |
| What type of stains are there | acidic and basic |
| Basic stains | are positive ( catatonic) and stain bacterial cells because of negative cell wall, |
| acidic stains | are negative and stain slide because of its positive charge |
| simple stain | uses acidic or basic stain to only add contrast |
| differential stain | uses multiple stains to differentiate between different cells |
| Methylene blue | basic positive charge, blue |
| Safranin | basic, positive charge, pink-red |
| Crystal violet | basic, positive charge, used in gram stain as primary stain |
| Carbol Fuchsin | Basic, Positive charge |
| Nigrosin | acidic, negative charge, black |
| India ink | acidic, negative charge |
| Congo red | acidic, negative charge, red |
| what does single simple stains do to specimens | allows morphology and arrangement to be visible |
| What do single acidic stains do to specimens? | allows morphology and arrangement to be visible for cells that are too delicate to heat-fix |
| What does differential staining consist of? | Primary stain, decolorizer, and counterstain |
| What is the differential component | the decolorizer |
| what stains all the cells | primary stain |
| what stains cells a different color | counterstain |
| Gram stain | differential stain for bacterial cell walls (grams-positive vs gram-negative) |
| capsule stain | differential stain |
| acid fast stain | differential stain |
| endospore stain | differential stain |
| Steps to preparing bacteria on slide | Add DI H2O to slide, add bacteria to slide using inoculating loop, heat-fix/air dry: a. kills bacteria, adheres bacteria to slide, and makes cells more permeable to stain |
| what does a typical staining procedure involve | 1. adding reagents to heat fixed slide one at a time for a specified number of time 2. rinsing reagent off slide after each addition 3. blot drying slide to bibulous paper after staining and before viewing under microscope |
| What does gram staining consist of? | 1. Primary stain using crystal violet, 2. Mordant: grams iodine, 3. decolorizer using ethanol, and 4. counter stain using safranin |
| E. coli | gram negative baccili |
| Salmonella | gram negative baccili |
| Pseudomonas | gram negative baccili |
| Bacillus | gram positive bacilli |
| Clostridium | gram positive bacilli |
| Listeria | gram positive bacilli |
| Neisseria | gram negative cocci |
| Staphylococcus | gram positive cocci |
| Streptococcus | gram positive cocci |
| Gram positive cell wall | thick layer of peptidoglycan, stains purple |
| Gram negative cell wall | thin layer of peptidoglycan but second membrane layer of LPS |
| Capsule staining | differentiates based on the absence or presence of a capsule |
| What does capsule staining consist of? | 1. Primary stain: Crystal violet, 2. decolorizer: 20% copper sulfate 3. 205 Copper sulfate, stains purple |
| Acid Fast stain | differentiates based on presence of mycolic acid in cell wall, Classifies bacteria as acid fast or non acid fast |
| What does an Acid Fast stain consist of? | 1. Primary Stain: Carbol Fucshin + moist heat 2: decolorizer: Acid alcohol 3. counterstain: methylene blue |
| What color does acid fast bacteria stain | red |
| What color does non-acid fast bacteria stain | blue |
| Endospore stain | differentiates based on presence or absence of an endospore, endospore formers or non-endospore formers |
| What does endospore stain consist of | primary stain: malachite green, decolorizer: DI H2O, counter stain: Safranin |
| What color does an endospore former appear as | green center and red background |
| What color does an non-endospore former appear as | red |
| Biological Culture media (media singular) | any substance that contains the necessary components for the growth of bacteria/organism |
| Bacteriological culture | any substance that is specifically formulated to include the requirements that promote the growth of bacteria |
| What are they different types of bacteriological culture medias? | 1. solid: Agar slant, agar deep, and agar plate 2. liquid media: broth culture, 3. semisolid: less dense than solid and less agar |
| What are the four types of bacteriological media? | General purpose, differential media, enriched, and selective |
| General purpose | contains necessary components to promote the growth of non-fastidious bacteria. |
| Selective media | contains one or more reagents that selects for a specific type of bacterial cell by inhibiting growth of some and promoting the growth of another |
| differential | contains one or more reagents that differentiates between different types of bacterial cells |
| Enriched media | contains component that promotes the growth of fastidious bacteria |
| What does a solid agar slant media culture allow | increases surface to maximize amount of growth. |
| What does an agar slant media contain | H2O, Agar, peptone, NaCl |
| What does an agar deep media contain? what does it do? | H2O, Agar, peptone, NaCl, creates an O2 gradient |
| What does an agar plate media contain? | H20, Agar, glucose, NaCl |
| What does an broth media contain | H2O, Glucose, peptone, NaCl |
| Subcultures | colonies created from taking parental colonies and inoculating them in in a sterile media, incubated and formed daughter cells |
| what bacteria was used in the culture transfer techniques? | Serratia marcescens |
| isolation of pure colonies methods? | T-streak and quadrant streak |
| In the isolation of pure colonies, what color did ecoli appear | white |
| In the isolation of pure colonies, what color did s. marcescens appear | red |
| viable plate count was used for | determine number of viable bacteria in broth culure |
| What was the process of serial dilution | used 7 broth cultures labeled 10^1-7 and 3 agar plates labeled 10^6-8, 1 ml of solution placed in vial, vortexed and placed in next vial. .01 ml from 10^5-7 placed in petri dishes, incubated at 30C |
| serial dulutions | decreases number of bacteria in each tube so that isolated colonies may form |
| CFU | Colony forming units |
| How is CFU calculated | # viable colonies/dilution factor |
| What does CFU require | 30-300 colonies |
Created by:
Acrob89
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