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Molecular Cell Bio Lab > Exam 2 > Flashcards

Exam 2 Flashcards

(59 cards)

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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)

Study These Flashcards
A

Pipette old media out and replace it with new media

25
2 factors to know its time to subculture:
-Cell density -Exhaustion of cell medium
26
cells becoming accustomed to new environment
Lag Phase
27
exponential cell growth (reproduction)
Log Phase
28
production of new cells is equal to death of cells
Stationary Phase
29
nutrients exhausted, waste has built up; death of cells exceeds production of new cells
Death Phase
30
Subculture at what percent confluency
80 to 90 percent
31
Medium exhaustion is indicated by:
drop in pH
32
Adherent cells must be detached from surface of the culture vessel via this method (in our lab)
Enzymatic dissociation
33
Protease; enzyme that digests protein Used for dissociation of cells from vessel surface
Trypsin
34
Used as a wash instead of water since water may kill cells
Phosphate buffered saline, or PBS
35
Inhibits trypsin, preventing extended trypsin activity that can kill cells after trypsinization
FBS
36
Lab 6 Reagents (Subculturing)
Trypsin PBS FBS
37
the component of the cytoskeleton that allows movement of cells and cellular processes
Actin
38
Type of actin used in Lab 7
Filamentous or F actin
39
Creates chemical cross-links between free amino groups in proteins (fixes cells)
Paraformaldehyde
40
Detergent solution used to permeabilize the membranes of living cells
Triton X-100
41
High-affinity F-actin probe conjugated to a red-orange fluorescent dye
Rhodamine phalloidin
42
Binds F-actin with high selectivity
Phalloidin
43
Fluorescent red-orange color
Rhodamine
44
Family of blue fluorescent dyes used to stain DNA (visualize nucleus)
DAPI
45
DAPI cannot be used with:
living cells
46
Lab 7 Protocol
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
cell viability indicator which can be used with living cells
alamarBlue
48
color of unhealthy cells
blue
49
color of healthy cells
pink
50
cell viability correlates with:
reducing power
51
Two variables affect the response of cells to alamarBlue:
1. Cell density 2. Incubation time
52
a technique for the visualization of proteins and peptides in cells using biomolecules capable of binding the protein of interest
Immunocytochemistry
53
the biomolecule is an _______ linked to a __________
antibody; reporter
54
4 steps of immunocytochemistry
1. cell seeding 2. immunostaining 3. imaging 4. image anlaysis
55
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
Direct ICC method
56
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
Indirect ICC method
57
Primary antibody species in Lab 9
Mouse
58
Secondary antibody species in Lab 9
Goat
59
a bright green fluorescent dye in Lab 9
Alexa Fluor 488

Molecular Cell Bio Lab (2 decks)

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