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XIO > Eating Habits - Lab > Flashcards

Eating Habits - Lab Flashcards

1
Q

Don’t place dirty slide in this beaker / tray.

A

cleaning solution

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2
Q

discarding contaminated materials

A

Remove all labels from tubes, placing them in appropriate racks on side of the room as shown

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3
Q

petri dishes with cultures

A

and contaminated swabs go in the biohazard bags

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4
Q

Dispose of used glass pipettes

A

in the discard pans on the side desk or placed on your desk. At the end before you leave lab, transfer all pipettes in one pan.

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5
Q

Don’t discard bulbs and pumps

A

in the pan

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6
Q

Before storing microscope back

A

replace the low objective lens into working position. The microscope is always stored with the lowest power objective lens locked in. Place plastic cover to avoid dust on the microscope

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7
Q

Condenser

A

focuses light on the specimen

  • is a lens system located below the stage
  • it doesn’t affect magnification
  • may include mirrors and prism to deflect light
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8
Q

Iris diaphragm lever

A

adjust light intensity for best contrast

- is part of the condenser system, below the stage

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9
Q

Coarse knob

A

Coarse knob: allows for gross focusing

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10
Q

Fine focus knob

A

Fine focus knob: allows for fine tuning of the focus

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11
Q

Head

A

Head: supports the objective and ocular lens system

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12
Q

Ocular lens

A

lens in each eyepiece; lens closest to the eyes
provides additional 10 x magnification when used with objective lens
monocular vs binocular microscopes (?)

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13
Q

Objective lens

A

adjustable lenses on the revolving nosepiece
normally used to change magnification of a specimen
built to minimize distortion in specimen image / color
4 types of objective lenses on microscope (scanning, low, high and oil immersion)

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14
Q

Total magnification

A

times 10

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15
Q

Resolving power - aka resolution - (2)

A

refers to ability of lens to distinguish between 2 points, a specified distance apart

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16
Q

Resolution is dependent on

A

quality, specimen prep, numerical, wavelength, white light

quality and type of lenses the magnification
how the specimen has been prepared
numerical aperture of lens (i.e. ability of the lens to gather light)
- larger the numerical aperture, better resolution
wavelength (?) of the light source; can limit resolution at higher magnification
shorter the wavelength of light used, higher the resolution
white light used in compound microscope limits resolution of structures smaller than 0.2μm

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17
Q

Working Distance (your objective)

A

refers to the space between slide specimen and objective lens - higher the magnification, shorter is working distance

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18
Q

Usefulness of a microscope depends on degree…

A

of magnification and resolve

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19
Q

Light or bright field microscope

A

easy to use, can recognize cells but not finer details. cells lack contrast with water.

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20
Q

Contrast

A

difference in intensity between object and it’s background

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21
Q

100x objective lens - how it works (drown)

A
  • oil displaces air, avoids refraction - oil increases numerical aperture (?), thus improves resolution
    because more light rays are gathered into lens to produce image
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22
Q

Refraction

A

prevents light from entering the small opening of the high powered objective lens

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23
Q

Refractive index

A

is measure of the velocity of light as it passes through medium

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24
Q

Phase contrast

A
  • contains special condenser to enhance contrast when viewing object - permits examination of internal structures- good for viewing objects without staining (as cilia and flagella)
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25
Q

Dark field

A

objects looks bright against dark background- contains special condenser with opaque disc to block light

  • good for viewing motility of organisms and
  • examining microbes that cannot be stained or they get distorted by staining procedures
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26
Q

Fluorescence microscope

A

normally use to visualize organisms that fluoresce

- often cells are treated with fluorescent dyes then observed for fluorescence

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27
Q

Electron microscope

A

uses electromagnetic lenses (?), electron and fluorscent screen to observe specimen instead of glass lenses and lightwavelength of electrons is approximately 1000x shorter than visible light
much higher resolution is obtained and greater detail can be observed.

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28
Q

Drawbacks to electron microscope

A

Lenses and processed specimen must be enclosed in a vacuum unit (air absorbs electron, expensive. Processing increases artifact chances and cannot be used to study living organisms

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29
Q

staining

A

contrast and light, internal and external structures, identification

provides contrast and increases resolution for light microscope- allows visualization with ease- reveals internal and external structures- aids in classification and identification

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30
Q

smear

A

smear thick - trouble seeing individual cells smear thin - you may find no organism too much stirring - disrupts cell arrangement

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31
Q

fixing (pipes)

A

adherence of microorganism to slide - accomplished by either heat or chemicals ( cold alcohol acetone mixture)

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32
Q

advantages of fixing

A

fixing inactivates the microorganisms- preserves major parts with minimum distortion - prevents specimen slipping in subsequent stain application and washing steps - makes subtle alterations so that organisms more readily accepts stains

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33
Q

mordant - purpose and what its made of

A

Fixes the dye. Iodine. an additive added to dye solution, to intensify the stain- it binds to the dye and makes it less soluble

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34
Q

advantages of mordant

A

like, coat, see
- it increases affinity of stain for biological specimen, stain is retained better- coats certain structures, so that it’s easier to visualize following staining

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35
Q

stains are made of what substance..

A

salts composed of positive and negative ion; only one part is colored. Chromophore: colored part of a stain

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36
Q

Basic (cationic) dye

A

positive part is colored - methalyne blue, crystal violet, malachite green

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37
Q

Acidic (anionic) dye (travel)

A

negative part is colored, sudan red, india ink

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38
Q

Simple stain

A

involves application of a single dye to a smear. highlights entire organism; shape and basic structures are visible, including arrangement of cells - are of relatively less value in diagnostic bacteriology

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39
Q

Differential stain

A

involves application of two or more dyes together or separately to a smear. mainly used to distinguish one group of bacteria from another- various dyes reacts with different bacterial structural components

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40
Q

Special stains - types FFN

A

FFN
3 types of special stains:
A. Negative stain (example: capsule stain) B. Flagellar stain C. Fluorescent stain

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41
Q

3 types of differential stain

A

gram, acid fast and spore stain

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42
Q

special stain

A

dye or color isolates specific structure of microbe.

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43
Q

negative stain uses what type of dye

A

on bacteria, uses anionic dyes. - chloroform.

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44
Q

bacteria is repelled by

A

acid. remains unstained.

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45
Q

halo around cell is…

A

negative acidic, likely the capsule

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46
Q

advantages of negative

A

get an idea of size and shape, and evidence of the capsule

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47
Q

what part gets stained in basic stain

A

the cell itself gets stained

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48
Q

chromatophores

A

part of plasma membrane. light reflecting cells.

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49
Q

euglena

A

looks like snap peas, has flagella. algae, eukaryote. bright green.

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50
Q

paramecium

A

dirty brown, eukaryote

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51
Q

stains (made of)

A

dyes, salt positive and negative ion

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52
Q

center of cheek stain dark part is

A

nucleus DNA

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53
Q

chromophore

A

positive part, blue part attracted to negative charge, DNA.

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54
Q

DNA negative part gets dyed due to

A

phosphate groups

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55
Q

gram negative color

A

pink and red

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56
Q

gram positive stains..(color)

A

peptioglycan, purple

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57
Q

decolorizing (nail polish +)

A

Alcohol/acetone. critical step. washes off stain. gram positive, water sucked out.

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58
Q

spore stain is a __stain

A

differential stain - can be positive or gram negative

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59
Q

gram positive rods..(ex)

A

make spores. only clostridium and bacillus anthracis.

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60
Q

use oil immersion lens with..

A

gram stain. no coverslip needed

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61
Q

acid-fast staining steps (car steam)

A

you’re mixing so it’s called mixed stain. primary dye - carbolfuscia dye on top of paper towel. heat with paper towel is fixing - steam acts as mordant. keep adding more carbonfuscia as you heat it. counterstain methalyne blue

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62
Q

real and virtual image

A

objective to real to ocular, ocular to virtual

The objective lens magnifies the specimen to produce real image that is projected to ocular lens . The real image is magnified by ocular lens to produce the virtual image.

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63
Q

virtual image is what position

A

upside down

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64
Q

stain used for cheek cell

A

alwasys simple stain, one dye

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65
Q

blue dye disassociates (eyes)

A

negative and positive. chromatophore is positive part, dye is attracted to negative part.

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66
Q

cheek cell is

A

eukaryotic cell

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67
Q

dark dots and rod shapes in cheek stain

A

are bacteria - prokaryotes

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68
Q

endspore stain is what type of stain

A

differential stain

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69
Q

advantages of gram staining

A

fast - gonnorhea, clues - strep pneumonia, treatment

it’s a fast and useful tool ex: urethral secretion (neissan gonorhea - only gram - diplococci) provides clues during the identification (lungs - strep pneumonia) for antimicrobial treatment (gram - like syphillis and TB can’t be treated)

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70
Q

disadvantages of gram staining

A

disease not distinct, can’t identify infection, stain poorly, need new cells

often, disease causing bacteria do not have distinct stain characteristics - the stain is not specific enough to diagnose the cause of most infection - some bacterial cells stain poorly or not at all; freshly grown bacteria is preferred. aged or old cells = misleading results

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71
Q

endspore staining process

A

primary stain - malachite green (basic dye). mordant = steaming (softens keratinized spore coat). wash and add secondary stain = safranin (basic dye, counter stain). Steaming needs to be done properly or it will be a light stain.

72
Q

endospore - green is the..

A

spore released from mother cell

73
Q

red endospores

A

mother spores (vegetative cells)

74
Q

dye that can’t enter mother cells

A

malachite green

75
Q

endospore staining bacillus anthracis

A

spores and vegetative cells. bacillus gram stain will always look like glass beads.

76
Q

spores only made by gram..

A

positive

77
Q

acid fast stain used because..

A

it’s another differential staining used for bacteria which do not readily stain by the Gram’s reaction although they belong to Gram positive group

78
Q

bacteria staining in acid fast (acidic beans)

A

Staining is facilitated by heat and phenol (present in the primary stain). Once stained, these cells are able to retain the stain because of the waxy lipid mycolic acid cell wall that are not easily decolorized by acid alcohol, so they appear red

79
Q

non-acid fast cells in acid fast stain

A

non acid fast cells easily decolorize and can be counterstained with another dye (methylene blue) and they appear blue

80
Q

acid fast procedure (blue car oil)

A

primary stain carbolfuschin. Apply mordant, heat and phenol (from the primary stain) by gently heating. use paper towel. STEAM IS THE MORDANT. decolorizer is acid alcohol. counterstain methylene blue. observe using oil immersion lens

81
Q

Acid fast organisms - which colors stain what

A

acid fast is stains red; non acid fast organisms stains blue

82
Q

non acid fast organisms (me)

A

stains blue

83
Q

what type of staining is acid fast?

A

differential staining

84
Q

we used _ bacteria in acid fast

A

gram +. don’t stain with gram stains, due to mycolic acid.

85
Q

in acid fast cells w/ mycolic acid, (car)

A

retain the carbolfuscion

86
Q

cord factor in acid fast

A

helps the pathogen to escape from host defense and being destroyed. it is a virulent factor.

87
Q

cord factor is made of

A

secreted protein and it sticks cells together. individually shaped cells are rod shaped.

88
Q

2 important bacteria used in acid fast in lab

A

m. tuberculosis and hansen’s disease (leprosy) called micobacterium leprae. Mycobacterium and Nocardia sps.

89
Q

cells w/ mycolic acid (acid fast) (1st car)

A

will retain primary stain, carbolfuschion

90
Q

no mycolic acid - acid fast stains what color

A

blue

91
Q

identifies TB

A

acid fast

92
Q

importance of sterile transfer techniques (newport)

A

not contaminated, protect me, isolate culture

keep bacterial specimen being studied, free of contaminants from environment; this is critical when a pure culture is desired
to prevent culture from contaminating you or others; this ensures to help yourself and others safe
to isolate pure culture of microbes
prevents observing features of microbes not considered as part of your experiment (contaminants are kept out); enables us to study metabolism of a particular organism

93
Q

precautions during sterile transfer

A

label, cool loop, no aerosols, high above flame, gently dry, don’t touch anything after sterile, Disinfect, no books

Carefully label all media, tubes and culture
Use a loop which is sufficiently cooled; don’t wave to cool
Avoid aerosols: loop has heavy load of residual microbes remaining
- hold loop high above flame
- gently dry it, before dropping to red hot area of flame
Keep transfer implements sterile
- do not touch any surface with it once flame sterilized
Disinfect work area before and after, remove unnecessary books and papers

94
Q

petri plates are labeled on

A

the bottom in case they are dropped and lid comes off

95
Q

petri dishes are placed in the incubator…

A

upside down due to water evaporation

96
Q

streak plate

A

widely used to obtain single colonies, which represents a pure culture of the bacterial cells.

97
Q

cultures used in streak plate (selene)

A

s. aureus and Serratia marcescenes

98
Q

common errors w/ streak plate (folsom)

A

fresh cells, pass twice, edge to edge, loop verticle, shake tube
Getting fresh cells on loop (from original culture tube) before streaking the 2nd and 3rd section
2. Failure to pass through previous section two times
3. Failure to go from edge to edge in each quadrant
4. Holding loop vertically; it’ll gauge agar
5. Forgetting to shake the tube, when obtaining cells for streaking in the first quadrant.

99
Q

differences in streak/spread plate and pour plate

A

spread plate - separated colonies are on agar surface and use inoculating loop to pick up cells from isolated colonies. pour plate - separated colonies are in between agar layer and use inoculating needle to stab agar and pick up cells. don’t dig.

100
Q

why are some colony sizes smaller when cells are closer? (SNW)

A

SNW

Due to competition for space, nutrients and metabolic waste accumulation that could be inhibitory to other bacteria

101
Q

emb agar - carbon source

A

lactose sugar, fermented, generates acid.

102
Q

emb agar - peptones

A

hydrolyzed proteins, source of nitrogen and amino acids

103
Q

emb agar - eosin dye is the…

A

differential factor

104
Q

emb agar - methlyne blue - which factor

A

selective factor, only supports growth of gram - rods.

105
Q

emb agar - extract

A

provides vitamins and essential nutrients

106
Q

emb agar process

A

for isolation, detection and confirmation of intestinal bacteria.

107
Q

bacteria used in emb

A

e.coli and enterobacteria.

108
Q

green sheen in emb

A

e.coli. eosin precipitates on colonies under acidic medium due to lactose fermentation. stable acids made, ph lowers. eosin dye precipitates turns green

109
Q

red emb

A

enterobacteria, eosin dye doesn’t precipitate bc lactose sugar ferments and forms unstable acids. fish eye colonies.

110
Q

Biochemical basis of selectivity and differential ability of EMB media (analise-no positive)

A

Eosin Y and Methylene Blue is differential and selective

selective because Eosin methylene blue agar contains aniline dyes that don’t allow growth of gram positive. Large amounts of acid from outside fermenters. Cause dye to precipitate on the colony surface producing a green color and pink.

111
Q

why is emb a complex media?

A

Because it uses different dyes to rule out different bacteria growth. This makes it a complex media because it can do several things at once.

112
Q

mordant in gram stain is

A

Iodine is as a mordant so dye can’t be removed easily

113
Q

decolorizing in gram stain (gina)

A

closes the pores in the cell wall and prevents the stain from exiting the cell.

114
Q

mordant in acid fast (melt)

A

heat, allows the primary stain to penetrate the waxy mycolic acid layer. The heat will prevent the cells from being destained using acid-alcohol

115
Q

decolorizer in acid fast does what (clean)

A

strips the stain in non acid fast cells. won’t enter the cell wall of acid-fast bacteria

116
Q

fixing in endospore stain

A

penetrates endospore, dye goes into peptidoglycan. heat is the mordant.

117
Q

primary stain in endospore stain driven into cells with…

A

heat

118
Q

counterstain in endospore stain binds to (poly)

A

lipopolysaccharide layer - gram -

119
Q

treponema palladium

A

(syphilis) doesn’t stain by gram stain or acid fast

120
Q

green sheen on agar plate is always..

A

e. coli

121
Q

oil immersion use

A

40X objective, focus and center the object
Lower stage and put a drop of oil on the slide
Change to 100X objective
Raise the stage using the coarse adjustment knob until objective lens touches the slide and the immersion oil on the specimen
Look through the eyepieces and lower the stage slowly, using the coarse adjustment knob
using the coarse knob until you see the specimen and then use the fine focusing knob
You may need to adjust using the fine focusing knob.

122
Q

resolution at high magnification

A

increase light intensity

Aperture size of the objective lens decreases with increased magnification; this allows less light to enter the objective lens. In such situation, with high magnification you may need to increase light intensity to obtain a good image of the specimen

123
Q

capsule stain - negative stain

A

distinguishes capsule from bacterial cell. The capsule stain uses acidic stain and a basic stain to detect capsule production

124
Q

advantages of negative stain

A

bacteria are not heat fixed so they don’t shrink, and some bacterial species resist basic stains (Mycobacterium) and one way they can be visualized is with the negative stain

125
Q

what happens in gram positive in gram staining

A

cell wall collapses and crystal violet trapped

126
Q

starch hydrolysis - exoenzymes

A

excreted out of cell, they break down extracellular substances, ex. amalyse.

127
Q

starch hydrolysis - endoenzymes

A

are enzymes used inside metabolic reactions. ex. endoenzyme ex. glycolic pathway and catalase.

128
Q

starch hydrolysis - process (potatoes)

A

use iodine to detect intact hydrolysis. iodine + intact starch = dark colored (blue). amalyse negative. iodine starch hydrolysized = halo around colonies. means it’s amalyse positive (cells secrete amalyse).

129
Q

starch hydrolysis detects which enzyme

A

a-amylase

130
Q

catalyse

A

Catalase is an enzyme produced by many microbes to neutralize the toxic effect of hydrogen peroxide. It breaks down hydrogen peroxide to generate water and oxygen. Oxygen release from the peroxide molecule is observed as bubble. Catalase presence can be detected by adding hydrogen peroxide directly onto bacterial culture tube or alternatively onto a dry smear of bacterial cell transferred to a slide

131
Q

results of catalyse and peroxidase

A

if you don’t see bubbles, absence of catalyst and peroxidase

132
Q

bubbles in catalyse

A

catalyse positive

133
Q

catalyse enzyme is a

A

endoenzyme

134
Q

what is generated in gas pak

A

h20 gas and CO2 generated inside chamber. can be noticed as moisture inside.

135
Q

anaerobic growth in gas pak

A

due to existing spores

136
Q

anaerobic bacterial growth can be cultivated (thyroid)

A

thiogycellate medium (absorbs dissolved 02) makes it unavailable for bacteria. physical method of heating followed by cooling, inoculating and tight screwing of cap, aneorobic jar w/ gas pak

137
Q

tsi - entobacterium - color change due to..

A

acid formation

138
Q

tsi - e.coli

A

use all 3 sugars. (slant) growth (butt) acid and gas. acid and gas production. yellow from fermentation and increases the acid ph. medium has been pushed up because gas was generated during fermentation.

139
Q

tsi - enterobacteria (instestines)

A

use all 3 sugars. (slant) growth (butt) acid and gas. intestinal bacteria as well, uses glucose first, then shift to use lactose and sucrose and it fermented, producing acid and gas.The medium has been pushed up because gas was generated during fermentation.

140
Q

tsi - proteus

A

(slant) growth (butt) growth. it uses glucose, but it cannot use lactose or sucrose. So it used amino acids of the peptones, the ammonia released and forms ammonium hydroxide. The blackening is due to anaerobic respiration and iron sulfide.

141
Q

blue on endospore slide is

A

endospore

142
Q

red on endospore slide is

A

bacillus anthracis - vegetative cell

143
Q

capsule stain is done using___stain

A

negative

144
Q

emb test uses what types of medium?

A

selective and differential medium

145
Q

(ex of selective media) thayer martin media used for

A

the isolation of neissan gonhorrea. gram -. This is selective medium.

146
Q

(ex of selective media) sabaroud’s is for..

A

fungi. it contains low ph, high salt and sugar. - inhibitory

147
Q

emb stands for

A

eosin methalyne blue

148
Q

emb uses what method?

A

streak plate

149
Q

in starch hydrolysis test, we are looking for..

A

amalyse. Amalyse breaks down starch to glucose units, then glucose is absorbed by bacterial cells.

150
Q

incubate plate at 37 degrees C

A

emb

151
Q

after you heat EMB, you use

A

iodine to detect intact or hydrolized starch. iodine reacts with intact starch to form dark color (washing machine). if dark, no amalyse.

152
Q

gram stain - primary and counterstain and mordant, decolorize

A

primary - crystal violet, counter - safranin, mordant - iodine, Decolorize - alcohol: acetone

153
Q

acid-fast - primary and counterstain and mordant

A

primary - carbolfuscion, counter - methalyne blue, mordant - heat. decolorize - acid alcohol

154
Q

spore stain - primary and counterstain and mordant

A

primary - malachite green, counter - safranin, mordant - heat. no decolorizer, primary stain washes off with water

155
Q

gram stain principles

A

differential stain, violet and iodine enters both + and -, forms crystal and iodine complex, alcohol removes stain from gram - only. alcohol dehydrates and removes h20 molecule. peptido in gram + collapses and traps iodine crystal violet complex. gram - outer membrane is destroyed and crystal violet comes out of cell. alcohol really important.

156
Q

eosin is an ___dye

A

acidic

157
Q

tsi - pseudomonas (ramona)

A

it is an aerobic bacteria, so there is no growth in the butt. It uses glucose first, it cannot use sucrose or lactose, so it uses the peptone amino acids as a carbon source. The blackening is caused by small amino acids, sulfur amino acids can be cysteine or methionine. They get diaminated, amine group released as ammonia, combines with water in medium to form ammonium hydroxide, it trickles down medium turning it red.

158
Q

peritrichous

A

e.coli

159
Q

monotrichous (ramona)

A

pseudomonas

160
Q

ampitrichous (voltans)

A

spirillum volutans

161
Q

deep butt

A

creates anaerobic, no oxygen. only anaerobic can thrive (or fermenting)

162
Q

shallow slant

A

provides aerobic

163
Q

purpose of TSI

A

to determine enterics from non enterics

164
Q

tsi - glucose is the lowest..

A

concentration, usually preferred energy source

165
Q

fermentation (alcohol)

A

pyruvic acid becomes either alcohol or lactic acid, then becomes acidic, few atp made

166
Q

respiratory

A

glucose becomes pyruvic acid, then goes to kreb. pathway can be areobic or anaerobic

167
Q

sodium thiosulfate in TSI is (runner up)

A

alternate electron acceptor in aerobic respiration, then it becomes hydrogen sulfide. then combines w/ iron sulfate and becomes iron sulfide. blackening in butt.

168
Q

strawberry red due to

A

ph indicator, alkaline acid. high ph will turn it red.

169
Q

carbon source in TSI

A

sugar

170
Q

yellow is tsi

A

is from fermented sugar

171
Q

iron sulfate detects (hs)

A

hydrogen sulfide

172
Q

iron sulfide is what color?

A

black

173
Q

pseudomonas - respiration is…

A

aerobic bacteria

174
Q

black in pseudomonas (sis)

A

due to cysteine and methionine amino acids

175
Q

acidic

A

low ph

176
Q

alkaline

A

high ph

177
Q

gas in e coli due to

A

fermentation. both e coli and enterobacteria have growth in butt.

XIO (393 decks)

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