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Foundations Exam 2 > Microbiological Lab Techniques > Flashcards

Microbiological Lab Techniques Flashcards

1
Q

Purpose of microscopy

A

To visualize smallmicroorganisms (diagnostic microbiology)

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2
Q

Brightfield microscopy

A

Use of focused light to visualize small objects (objective x ocular lens = mag); example; wet mount

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3
Q

Darkfield microscopy

A

Scattered or reflected light to visualize object; visualization of thin bacteria (spirochete)

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4
Q

Fluorescent microscopy

A

One wavelength of light is used to cause another wavelength of light to be detected; fluorescent molecules target specific cell components

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5
Q

Electron microscopy

A

Electrons used to visualize; high mag; transmission = electrons through; scanning = electrons bounce

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6
Q

Purpose of direct examination

A

Visualize cells with minimal preparation

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7
Q

Wet mount

A

Direct visualization in water/saline; no processing (i.e. vaginal wet mount)

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8
Q

KOH

A

Usually 10% to dissolve proteinaceous non-fungal elements

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9
Q

Lactophenol cotton blue

A

Type of KOH added to non-specifically stain material (ex. fungal elements)

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10
Q

India ink

A

Type of KOH added to stain everything but yeast polysaccharide capsule (ex. cryptococcus)

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11
Q

Iodine

A

Stain organelles darker than cytoplasm (ex. parasite cysts)

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12
Q

Differential stain

A

Colored dyes that differentiate organisms or structures within organisms

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13
Q

Acid fast stain

A

Stains mycobacterium, nocardia, cryptosporidium

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14
Q

Fluorescent stains

A

Calcofluor white stain - interacts with cellulose and chitin in cell walls and fluoresces blue

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15
Q

Wright-Giemsa stain

A

differential used to stain parasites, intracellular bacteria, viral elements, and eukaryotic cells (polychromatic stain that contains azure B, methylene blue, and eosin Y; eosin ions stain basic structures orange/pink, others stain acidic structures blue/purple

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16
Q

Methenamine silver

A

Used to stain fungal elements and some bacteria in tissues

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17
Q

Trichrome stain

A

Used to detect fecal protozoa

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18
Q

Gram stain

A

1.) Heat fix
2.) Crystal violet solution - purple stain is positively charged; enters bacterial cell and complexes with anionic molecules
3.) Gram’s iodine - iodine acts as a mordant; interacts with crystal violet and forms a larger complex
4.) Gram’s decolorizer - alcohol dehydrates; peptidoglycan wall shrinks and captures crystal-violet iodine complex within cell
Gram + = thick cell wall doesn’t let crystal violet through
Gram - = thin cell wall allows stain through
5.) Safranin - red counter stain that stains whatever primary stain didn’t (Gram - )

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19
Q

Ziehl-Neelsen and Kinyoun stains

A

Same principle as gram stain but for mycobacterium; carbofushin (purple dye) added; alcohol washes what didn’t stain; methylene blue (green) acts as counter stain

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20
Q

Auramine-rhodamine

A

Fluorescent stain of mycolic acids

21
Q

Purpose of culture medium

A
  • select for growth of specific microorganisms

- differentiate amongst microorganisms

22
Q

Nonselective culture

A

Supports growth of many organisms

23
Q

Trypticase soy/broth agar

A

Digested soybean meal and casein; full of nutrients (supports growth of many organisms)

24
Q

Nutrient broth/agar

A

Peptone and beef extracts; full of nutrients

25
Q

Blood agar

A

Trypticase soy with sheep blood (enriched)

26
Q

Chocolate agar

A

Trypticase soy with lysed sheep blood (enriched)

27
Q

Thioglycolate broth

A

Sodium thioglycolate added to promote a reducing (anaerobic) environment; used to grow anaerobic bacteria and can be used to differentiate oxygen usage

28
Q

Sabouraud dextrose agar

A

Growth of fungi

29
Q

Selective culture; differential

A

Supports growth of specific organisms usually by limiting growth of others; differentiation of organisms in a mixture

30
Q

MacConkey agar

A

Selective for gram- ; differential for lactose fermentation

31
Q

Mannitol salt agar

A

Selective for gram+ Staphylococci; differential for S aureus (mannitol fermentation)

32
Q

Lowenstein-Jensen agar

A

selective for mycobacteria species

33
Q

Eosin-methylene blue agar

A

Selective for gram- ; differential for lactose fermentation

34
Q

Hektoen enteric agar

A

Selective for gram - ;differential for hydrogen sulfide production (black) and lactose/sucrose/salicin formation (pink)

35
Q

Specialized culture

A

Ingredients support the growth of specific organisms

36
Q

Thayer martin

A

Neisseria gonorrhoeae

37
Q

Buffered charcoal yeast extract (BCYE) agar

A

Legionella

38
Q

Thiosulfate citrate bile salts sucrose (TCBS) agar

A

Vibrio species

39
Q

Regan-Lowe agar and Bordet-Gengou agar

A

Bordetella pertussis

40
Q

Purpose of biochemistry in lab techniques

A

Differentiation of bacteria based on varying biochemical pathways

41
Q

Catalase test

A

Bubbles = + catalase; catalase enzyme functions as part of the antioxidant defense response; breaks down H2O2 to O2 and 2H2O

42
Q

Oxidase test

A

Purple = positive (before 30 secs); some bac use O2 as final electron acceptor (reduce O2 to form H2O via cytochrome oxidase complex)

43
Q

Antibiotic sensitivity test (Kirby-Bauer, Disk Diffusion)

A

Wells with increasing amounts of antibiotics are inoculated with clinical sample; resistant = will grow, sensitive = no growth; first well in an antibiotic series with no growth is the minimum inhibitory concentration

44
Q

Purpose of immunology in labs

A

Use of immune components (usually antibodies) to differentiate microorganisms

45
Q

Examples of immunology in lab

A

Latex agglutination, immunofluorescence, ELISA, western blot, and others

46
Q

Latex agglutination

A
  • Latex bead solution (with antibody coating) added to flat surface
  • mix bacterial strain in
  • evaluate for precipitation (precipitate = +)
47
Q

Immunofluorescence

A

Use of antibodies that are conjugated to fluorescent molecules

48
Q

Purpose of eukaryotic cell culture

A

Grow intracellular bacteria and viruses; eukaryotic cells act as the culture medium for intracellular bac and viruses

Foundations Exam 2 (8 decks)

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