Micro Lab final

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electric cord

dont dangle from table so that a foot gets caught and is pulled off table.

stage

horizontal platform

ocular, objective, condenser

three lens of the brightfield

condenser

lens that collects and directs the light from the lamp

diaphram

regulates the amount of light that reaches the slide

adjusting knobs

fine adjustment knob and coarse adjustment knob

magnification

ocular 10 times the objective 10, 40, 100

resolving power

ability to completely separate two objects in a microscopic field

limit of resolution

0.2 um for light microscopes

parfocal

image remains in focus when changing from low power objective lens to high power objective lens

lamp life is extended

when opening diaphram fist instead of increasing voltage

oil immersion

oil has the same refractive index as glass and will increase resolving power by reducing light lost from refraction

brightfield microscope

allows light ot pass directly to the eye

bacteria

have simple cell structure that lacks defined nucleus and no nuclear membrane

peptidoglycan

unique molecule in bacterial cell wall

spirochete

motile, using axial filaments, which are a type of flagella that originate from both ends of the cell and wrap around the cell body

heterotrophic

"other" + "eating" - organisms which feed on other plants and animals, as oposed to autotrophs that use CO2

fungus

eukaryotic, nucleus, produce exoenzymes and absorb thier nutrients, lack tissue definition, have cell wall of chitin, propogate (multiply) by spores

morphological characteristics

structural, the form and structure of an organism considered as a whole

yeasts

unicellular, lack hyphae, multiply by budding or fussion or both

molds

multicellular

hyphae

microscopic intertwining filaments of molds

mycelium

mass of hyphae; macroscopic fungal colony

psuedohyphae

chains of buds of yeasts

asexual fungi

sporangiosopres, chlamydospores; does not involve the union of cell nuclei

sexual fungi

zygospores, ascopores, basidiospores; union of two parental nuclei followed by meotic division

eumycota

the fungal kingdom, "True fungi"

Saccharomyces cervisiae

yeast used in bread making and alcohol; blastospores

colony characteristics of molds

cottony, different colors, round or irregular

dimorphic fungi

characteristics of both mold and yeast

goals of aseptic technique

no unwanted organisms in culture, you are not contaminated, no contaminates are left afterwards

work area disinfected

destroys vegetative cells and viruses, but may not destroy endospores

vegetative

active bacteria, not endospores; cells involved in obtaining nutrients,not reproduction

endospores

resting, resistant, dormant structure formed inside some bacteria that protects form adverse environment; dehyrated with thick cells walls with extra layers

endospore staining

dyes do not penetrate the wall

Shaffer-fulton

malachite green is heat steamed, counterstain with safranin

survival of endospore

resisitant to heat, lack of water, and exposure to chemicals and radiation

loop or needle

is sterylized by inserting into bunsen flame until it is red hot

culture tube

mouth is flamed prior to and after inserting a loop or needle

petri plate

cover is held diagonally over the plate to protect the surface from any contaminates in the air

work area

should be cleaned with disinfectant before and after work

inoculated petri plates

incubated upside down to prevent moisture from condensing on the agar surface and spoiling or spreading inoculated organism

mixed populations

how bacteria exist in nature

pure culture

contains only a single kind of bacteria; able to study morphological, cultural and physiological characteristics

obtain pure cultures

streak plate and pour plate dilute bacteria to a point where a single cell gives rise to single pure colony

streak plate

economy and time make this method best

agar

becomes liquid at 85 and resolidifies at 42. Good gel stability at high temperatures; made from polysaccharide of algae

quadrant streak

technique that spreads oranisms around plate in a way that dilutes concetrations

two reasons plates are incubated upside down

water will spread organism, may inhibit growth

condesation on plate cover

can be avoided by cooling agar to 50 before pouring

goals of smear

adhere cells to slide and insure shrinkage does not occure; prepare thin

reasons for thin slide prep

to visualize individual cells, their arrangment and details regarding microstructures; stains better

simple staining

use of single stain to color bacterial cell

simple stains

methylene blue, basic fuchsin and crystal voilet are basic dyes

positive stains

bacteria are negatively charged

staining time

30 seconds to 2 minutes

pleomorphic, pleomorphism

irregularity of form, demonstrating several different shapes

metachromatic granules

distinct reddish-purple granules with cells that show up when the organisms are stained with methylene blue

palisade arangement

parallel arrangment of rod shaped cells "picket fence" (characteristic of corynebacteria)

capsule

slime layer, gycocalyx, protective role for certain bacteria like streptococcus pneumoniae, prevents phagocytosis by WBC, to attach to solid surfaces

shrinks capsule

heat fixing prior to staining

india ink

negative stain

Streptococcus pneumoniae

capsule producing to evade WBC phagocytosis in lungs

primary stain

for gram stain is crystal violet

differential stain

take advantage of the fact that cells or structures within cells display dissimilar staining reactions that can be distinguished by the use of different dyes

gram positive bacteria

retain crystal violet-iodine complex through decolorization with alcohol; appear purple

gram negative bacteria

alcohol removes the crystal violet iodine complex; counterstain of safranin makes bacteria appear pink

mordent

for gram stain is iodine; forms complex with crystal violet that is insoluble in gram positive cells

decolorization

alcohol leaches the dye-mordent complex from gram negative cells

gram positive cell wall

has thick peptidoglycan layer

gram negative cell wall

has thin petidoglycan layer

counterstain

for gram stain is safranin, to stain any gram negative cells in the smear

Mycobacterium smegmatis

acid fast bacteria that stains gram positive; representative of acid fast bacteria

gram positve

S. aureus

gram negative

E. coli

endospores

allow bacteria to survive environmental conditions that are not favorabe for growth; resistant to heat, radiation, acids, and many chemicals such as disinfectants

exosporium

protien coat that forms a protective barrier around the spore

low water content of spores

provides heat resistance for endospores

dipicolinic acid

forms gel that controls the amount of water that can enter the endospore, thus maintaining its dehydrated state

autoclave

121 degrees for 15-20 minutes to destroy endospores

heat

mordent in spore staining to facilitate the uptake of malachite green

Shaffer-Fulton method

utilizes malachite green to stain the endospore and safranin to stain the vegetative portion

Bacillus subtilis

endospore stain bacteria

acid fast

bacteria such as mycobacteria and some nocardia

mycolic acid

waxy material; complex lipid that is composed of fatty acids and fatty alcohols that have hydrocarbon chains up to 80 carbons in length; prevents routine stains

acid fast stain

diagnositc tool for Mycobacterium tuberculosis and Mycobacterium leprae (tuberculosis and leprecy)

Zeil-Neelsen method

carbolfushion mixed with phenol is heated, creates noxious fumes toxic to the eyes and mucous membranes

Kinyoun method

acid fast stain that is not heated, concentrations are increased for same results

acid-fast

carbolfuchsin mixed with phenol (for permeability), then decolorized with alcohol, counterstained with methylene blue

non acid fast

S. aureus

thioglycolate medium

test to determine the oxygen requirements by observing the nature of growth in the tube

parasitology

study of protozoans, parasitic worms (helminths), and parasitic arthropods such as lice, mites and ticks

sacodina

protozoa with locomotion by means of psuedopods

mastigophora

protozoa with locomotion by means of flagella

ciliata

protozoa with locomotion by means of cilia

sporozoa

nonmotile protozoa

according to motility

how protozoa are classified

trophozoite

motile form of protozoan, pathological form

cyst

after leaving the warm host, cools and curls, forms thick walls, becomes inactive

Entamoeba histolytica

representative sarcodina protozoan; moves by psuedopod

repesentative flagellate; mastigophora

Trichomonas vaginalis, Giardia lamblia, Trypanosoma gambiense

plasmodium

causes malaria

true

protozoans are classified according to their modes of locomotion

flagellate

mean of mobility for Giardia lamblia

False, cyst

laboratory identification of the parasitic protozoans is usually made from microscopic examination of the trophozoite stage.

true

specimens containing the parasitic trophozoite stage are more readily available for classroom study than those containing the cyst stage.

trophozoite

stage that cause pathological conditions in the host

flagella

Giardia is identified by this

tsetse fly

transmits african sleeping sickness

Giardia

found in humans and animals, especially beavers

plasmodium species

malaria

Entamoeba histolytica

dysentery with fluid feces

Typanosoma gambiense

drowsiness, coma and death

giardia lamblia

diarrhea, abdominal cramps; beaver fever

Ascaris lumbricoides

ova must live in soil for several weeks before able to infect humans

nematodes

parasitic round worms

Trichinosis

diagnosed by clinical symptoms, serology tests and muscle biopsy

Nectator americanus

burrow through the soft skin on the sides of the feet

Trichinella spiralis

females bear fully developed, viable larvae instead of depositing ova

Taenia

tapeworms

tapeworm infestation

result of ingestion of encysted larvae in the improperly cooked flesh of an intermediate host animal

measly beef

beef visibly infested with Taenia cysts

Taenia ova

tapeworm stage not infectious for humans

distal progottids

more mature than the proximal on the tapeworm

free living

nonparasitic protozoans

observable traits, phenotype

Color of colonies, number of colonies, distribution of colonies on the plate.

good choices for genetic transformation experiments

non toxic, reproduces quickly, single cell organism

LB amp and LB amp/ara

plates that the transformed organisms will be found

Genetically transformed cells

cells that have taken up the pGLO plasmid express the ampicillin resistance gene—these cells can survive on the plates which contain ampicillin.

determine success of transformation

The LB/amp (-) pGLO and the LB/amp (+) pGLO plates should be directly compared.

ampicillin

antibiotic that only the organisms that took the plasmid will be able to survive

arabinose

sugar that regulates the expression of GFP, green florescent protien

pGLO plamid

GFP, ampicillin resistance, arabinose gene regulation system

control plate

guide your interpretation of results in an experiment; usually a plate of your original without manipulation

bacterial lawn

so many bacterial on a plate that individual colonies are not present, looks like a complete covering of growth

protien that the plasmid encodes

source of the florescences

how to determine ampicillin resistance

take some of the bacteria growing on the LB plate and
streak them on an LB/amp plate.

arabinose sugar

turn on the expression of the GFP gene.

two factors that cause bacteria to glow green

The sugar arabinose turns on expression of the GFP gene by binding to a regulatory protein. When arabinose
is present, it facilitates transcription of the gene by RNA polymerase. Exposure to UV light causes GFP to
resonate, thereby giving off energy in the form of green light.

advantage of turning off a gene

Gene regulation allows for adaptation to differing conditions and prevents wasteful overproduction of unneeded proteins.

E. coli

gram negative rod, nonspore forming, ferment lactose and live in the colon of humans

S. aureus

gram positive cocci, cause of staph infections, golden color, commensal on skin, produce enterotoxin

coliform

commonly-used bacterial indicator of sanitary quality of foods and water; almost exclusively of fecal origin and their presence is thus an effective confirmation of fecal contamination; E. coli and Enterbacter aerogenes

positive result for E. coli

metallic green colonies

b. subtilis

gram positive rod, endospore forming, nonpathogenic, found in soil, flagellated

saprophytes

nonpathogenic bacteria that may be found in drinking water

Vibrio cholerae, Salmonella typhi, Shingella dysenteriae

pathogens in contaminiated water

E. coli

bacteria that indicates likelihood fecal contamination in water and the potential for serious disease

E. coli is a good indicator

occurs primarily in the intestines of of humans and warm-blooded animals and is not found routinely in soil or water; easily identified by microbiological tests; not as fastidous as the intestinal pathogens and survives a little longer in water samples

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