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Cellular and Molecular Biology > Exam 3: Ch 9 Culturing Cells & Microscopy > Flashcards

Exam 3: Ch 9 Culturing Cells & Microscopy Flashcards

(63 cards)

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1
Q

culturing

A

maintaining isolated cells in the lab under conditions that permit their survival and growth

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2
Q

3 advantages of cultured cells

A

grow cells of a single type

control experimental conditions

single cell easily grown into a colony of identical cells

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3
Q

clone

A

strain of identical cells grown from a single cell

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4
Q

immunoflurorescence microscopy

A

allows localization of specific proteins within fixed cells

provides a static image of their location

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5
Q

chimeric protein

A

protein of interest covalently linked to a naturally fluorescent protein

allows visualization of movements of the protein in live cells

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6
Q

organelle

A

membrane limited compartment of a cell

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7
Q

culture medium

A

nutrient rich liquid

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8
Q

cell-adhesion molecules

A

cell surface proteins that allow cells to bind to each other and to the ECM

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9
Q

primary cell culture

A

a culture where the cell-cell and cell-surface interactions are broken by protease and chelation

fibroblasts become predominant cell (they lay ECM)

finite life span

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10
Q

embryonic stem cells

A

can give rise to all tissues during development

can be cultured indefinitely

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11
Q

oncogenic transformation

A

cells that are able to grow indefinitely (happens spontaneously)

a culture of immortal cells like this is a cell line

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12
Q

flow cytometer

A

desired cell type labeled with an ab + attached fluorescent dye

cells flow past a laser beam that measures the emitted fluorescence to quantify # of cells of desired type in a mix

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13
Q

fluorescence activated cell sorter (FACS)

A

a flow cytometer that can analyze the cells and sort them

ex. purify T cells from others (T cells have CD3 and THy1.2(

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14
Q

apical

A

top surface

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15
Q

basal

A

bottom surface that contacts underlying ECM called basal lamina

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16
Q

Madin-Darby Canine Kidney (MDCK)

A

cells grown in special containers to study epithelial cellsq

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17
Q

hybrid cells called ______ produce abundant monoclonal antibodies

A

hybridomas

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18
Q

antibody

A

protein that binds to foreign molecules and tags them for elimination

made by memory B cells

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19
Q

B cells are made where and by what

A

in the bone marrow from stem cells

genes undergo recombination during maturation to increase variety of Ab genes

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20
Q

Ab are on the surface of _ cells

A

B

if antigen binds to the B cells, gets activated to make memory B cells

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21
Q

memory B cells

A

make Ab if bound to antigen

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22
Q

can a foreign molecule have multiple epitopes?

A

yes, reason for making polyclonal Ab

multiple B cells attach

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23
Q

polyclonal

A

multiple clones of B cells

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24
Q

monoclonal

A

one clone of B cells

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25
how to make a monoclonal Ab
inject mice with antigen, kill mouse at 2 wks, remove spleen, separate B cells w/ assay and choose Ab w/ best affinity
26
problem with monoclonal Ab
B cells stop dividing/Ab stop dividing eventually solution: hybridoma (cancer cell + B cell cross)
27
myeloma cell
immortal lymphocytes that are joined with a normal B cell to make a hybridoma
28
selection medium
medium that permits growth of only hybridomas
29
what can monoclonal Ab be used for
affinity chromatography immunofluorescence microscopy immunoblotting medicine
30
obejctive lens
lens closest to the speciment
31
projection lens
ocular or eyepiece
32
resolution
ability to distinguish btw two closely positioned objects limit for a light microscope is .2 micrometers
33
two types of microscopy
visible light fluorescence
34
visible light microscopy is ___ contrast
low increase by staining or optical tricks
35
phase contrast microscopy
increase contrast using a phase ring that reduces bent light direct light passes through an area of phase plate and transmits a small percentage of light depends on thickness of specimen and difference in refractive index
36
differential interference contrast microscopy (DIC)
increase contrast using a shadow effect with a condenser filter method of choice for visualizing small details and thick objects
37
dark field
no specimen = see blackness specimen sparkling in light
38
why stain?
live cells lack compounds that absorb light, and are invisible to a light microscope
39
light and electron microscopy use stains that...
cross-link most proteins and nucleic acids ex. formaldehyde cut into sections 50 micrometers thick
40
a molecule is said to be fluorescent if
it absorbs light at one wavelength and emits light at a specific and longer wavelength
41
fluorescent microscope staining
get higher quality image (tag important part in thick specimen) use UV light as excitation source for UV tags -- visualize emitted fluorescence living or dead cells
42
fluorochrome
fluorescent dye ex. fura-2 sensitive to Ca2+, SNARF-1 sensitive to H+
43
immunofluorescence microscopy can be used to detect specific _____ in fixed cells
proteins -- have Ab with fluorescent dye attached
44
immunofluorescence microscopy
Ab covalently linked to fluorochrome like Texas red or green fluorescent protein when bound to antigen, lights up
45
reporter gene
glow tells us where gene is active
46
lower E = ______ wavelength
longer
47
goat anti-rabbit
indirect immunofluorescence immunize a goat with Fc common to all rabbits to get better fluorescence upon binding
48
epitope tag
cDNA encoding a recombinant protein fused to a short sequence of aa when expressed in cells, the cDNA will generate the protein linked to the tag
49
2 problems with fluorescent microscopy
light emitted from above and below plane of focus gives blurry image to visualize thick specimens, serial images must be collected and reconstructed by computer (time-consuming)
50
zebrafish microscopy
have clear embryo fluorescence tagging used to see what grows where in growing embryo
51
deconvolution microscopy
collect into overnight and computer takes out blurry info
52
confocal microscopy
uses optical methods to obtain images from a specific focal plane and exclude light from others (otherwise image degraded) light through pinhole laser scanning spinning disk
53
focal plane
sliver of a thick specimen
54
laser scanning confocal microscopy (LSCM)
eliminate out of focus info using a laser to limit light multiple pics to make a stack of pics Z series = multiple planes... 3D image
55
total internal reflection fluorescence (TIRF)
visualize a thin focal plane like kinetics of microtubules and actin filaments
56
fluorescence resonance energy transfer (FRET)
used to test if 2 molecules are next to each other use 2 fluorochromes of different color where emitted light of 1st excites 2nd illumination of 1st fluorochrome produces 2nd light if nxt to each other
57
super resolution microscopy is a subset of ________ microscopy
fluorescence
58
photo activated localization microscopy (PALM)
uses a type of GFP that only fluoresces when activated by a specific wavelength of light, different from its excitation wavelength look at 1 molecule at a time
59
strategy x
slightly different times of excitation and release
60
electron microscopy
electromagnetic lens focuses a high-velocity electron beam living material cannot be visualized
61
transmission electron microscopy
visualize things that have been negatively stained with heavy metal
62
cryoelectron microscopy
view biological specimens using TEM if the specimen is frozen ex. virus capsids
63
scanning electron microscopy (SEM)
view surfaces of unsectioned metal-coated specimens limited by thickness of metal coat

Cellular and Molecular Biology (34 decks)

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