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Molecular Cell Bio Lab > Exam 2 > Flashcards

Exam 2 Flashcards

1
Q

the percentage of the surface of a culture dish that is covered by adherent cells

A

confluence

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2
Q

transferring cells to a new vessel with fresh growth medium to provide more room for continued growth.

A

subculturing

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3
Q

After the first subculture, the primary culture becomes known as a:

A

cell line/subclone

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4
Q

bipolar or multipolar, have elongated shapes, and grow attached to a substrate

A

Fibroblast-like cells

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5
Q

polygonal in shape with more regular dimensions, and grow attached to a substrate in discrete patches

A

Epithelial-like cells

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6
Q

spherical in shape and usually grown in suspension without attaching to a surface

A

Lymphoblast-like cells

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7
Q

monolayers on an artificial substrate

A

adherent

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8
Q

free-floating in the culture medium

A

Suspension

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9
Q

Generally used to grow large batches
Contain filtered caps to allow CO2 exchange

A

Flasks

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10
Q

Used for smaller batches
Individual control over the wells

A

Well plates

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11
Q

Generally used with bacteria culturing
Often solid medium is used

A

Petri dishes

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12
Q

Media/medium - 4 components

A
  1. Amino acids
  2. Inorganic salts
  3. Vitamins
  4. Sometimes antibiotics
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13
Q

Serum - 3 components

A
  1. Macromolecules
  2. Lipids
  3. Growth factors
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14
Q

Most normal mammalian cell lines grow well at this pH

A

7.4

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15
Q

Most mammalian cells grow best at:

A

37°C

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16
Q

Humid CO2 incubators control these 3 conditions

A
  1. CO2
  2. Humidity
  3. Temperature
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17
Q

provides an aseptic work area while allowing the containment of infectious splashes or aerosols generated by many microbiological procedures.

A

laminar flow hood

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18
Q

only protect the cultures from the worker and used if the cultures are harmless to you and the environment

A

clean benches

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19
Q

generally used to protect the cultures from workers and workers from the cultures

A

Biological safety cabinets (BSCs)

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20
Q

Sterile/aseptic techniques

A

-Wear gloves
-Spray everything with ethanol
-Keep hands inside hood
-Change pipette tips for new materials
-Don’t pour reagents directly from bottles/flasks
-Don’t uncover anything until its ready for use

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21
Q

the most common type of brain tumor

A

glioblastoma

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22
Q

Usually has poor prognosis and high mortality for glioblastoma due to:

A

its highly invasive behavior

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23
Q

Lab 5 Reagents (Changing media)

A

-DMEM
-FBS
-Penicillin-Streptomycin

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24
Q

Lab 5 Protocol (changing media)

A

Pipette old media out and replace it with new media

25
Q

2 factors to know its time to subculture:

A

-Cell density
-Exhaustion of cell medium

26
Q

cells becoming accustomed to new environment

27
Q

exponential cell growth (reproduction)

28
Q

production of new cells is equal to death of cells

A

Stationary Phase

29
Q

nutrients exhausted, waste has built up; death of cells exceeds production of new cells

A

Death Phase

30
Q

Subculture at what percent confluency

A

80 to 90 percent

31
Q

Medium exhaustion is indicated by:

A

drop in pH

32
Q

Adherent cells must be detached from surface of the culture vessel via this method (in our lab)

A

Enzymatic dissociation

33
Q

Protease; enzyme that digests protein
Used for dissociation of cells from vessel surface

34
Q

Used as a wash instead of water since water may kill cells

A

Phosphate buffered saline, or PBS

35
Q

Inhibits trypsin, preventing extended trypsin activity that can kill cells after trypsinization

36
Q

Lab 6 Reagents (Subculturing)

A

Trypsin
PBS
FBS

37
Q

the component of the cytoskeleton that allows movement of cells and cellular processes

38
Q

Type of actin used in Lab 7

A

Filamentous or F actin

39
Q

Creates chemical cross-links between free amino groups in proteins (fixes cells)

A

Paraformaldehyde

40
Q

Detergent solution used to permeabilize the membranes of living cells

A

Triton X-100

41
Q

High-affinity F-actin probe conjugated to a red-orange fluorescent dye

A

Rhodamine phalloidin

42
Q

Binds F-actin with high selectivity

A

Phalloidin

43
Q

Fluorescentred-orange color

44
Q

Family of bluefluorescentdyesused tostainDNA (visualize nucleus)

45
Q

DAPI cannot be used with:

A

living cells

46
Q

Lab 7 Protocol

A
  1. Fix with paraformaldehyde
  2. Permeabilize with Triton-x100
  3. Bind actin and stain with rhodamine phalloidin
  4. Bind and stain DNA with DAPI
47
Q

cell viability indicator which can be used with living cells

A

alamarBlue

48
Q

color of unhealthy cells

49
Q

color of healthy cells

50
Q

cell viability correlates with:

A

reducing power

51
Q

Two variables affect the response of cells to alamarBlue:

A
  1. Cell density
  2. Incubation time
52
Q

a technique for the visualization of proteins and peptides in cells using biomolecules capable of binding the protein of interest

A

Immunocytochemistry

53
Q

the biomolecule is an _______ linked to a __________

A

antibody; reporter

54
Q

4 steps of immunocytochemistry

A
  1. cell seeding
  2. immunostaining
  3. imaging
  4. image anlaysis
55
Q

One reporter labeled antibody is used
Quick
Not sensitive enough for most proteins as the number of present copies of the protein is too low to yield a strong signal

A

Direct ICC method

56
Q

Multiple binding of secondary antibodies to the same primary antibody
Increased sensitivity
Amplification of signal
Increased flexibility
More work and time
Risk of non-specific binding of the secondary antibody

A

Indirect ICC method

57
Q

Primary antibody species in Lab 9

58
Q

Secondary antibody species in Lab 9

59
Q

a bright green fluorescent dye in Lab 9

A

Alexa Fluor 488

Molecular Cell Bio Lab (2 decks)

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