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

Exam 3: Ch 9 Culturing Cells & Microscopy Flashcards

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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
Q

how to make a monoclonal Ab

A

inject mice with antigen, kill mouse at 2 wks, remove spleen, separate B cells w/ assay and choose Ab w/ best affinity

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26
Q

problem with monoclonal Ab

A

B cells stop dividing/Ab stop dividing eventually

solution: hybridoma (cancer cell + B cell cross)

27
Q

myeloma cell

A

immortal lymphocytes that are joined with a normal B cell to make a hybridoma

28
Q

selection medium

A

medium that permits growth of only hybridomas

29
Q

what can monoclonal Ab be used for

A

affinity chromatography

immunofluorescence microscopy

immunoblotting

medicine

30
Q

obejctive lens

A

lens closest to the speciment

31
Q

projection lens

A

ocular or eyepiece

32
Q

resolution

A

ability to distinguish btw two closely positioned objects

limit for a light microscope is .2 micrometers

33
Q

two types of microscopy

A

visible light

fluorescence

34
Q

visible light microscopy is ___ contrast

A

low

increase by staining or optical tricks

35
Q

phase contrast microscopy

A

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
Q

differential interference contrast microscopy (DIC)

A

increase contrast using a shadow effect with a condenser filter

method of choice for visualizing small details and thick objects

37
Q

dark field

A

no specimen = see blackness

specimen sparkling in light

38
Q

why stain?

A

live cells lack compounds that absorb light, and are invisible to a light microscope

39
Q

light and electron microscopy use stains that…

A

cross-link most proteins and nucleic acids

ex. formaldehyde

cut into sections 50 micrometers thick

40
Q

a molecule is said to be fluorescent if

A

it absorbs light at one wavelength and emits light at a specific and longer wavelength

41
Q

fluorescent microscope staining

A

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
Q

fluorochrome

A

fluorescent dye

ex. fura-2 sensitive to Ca2+, SNARF-1 sensitive to H+

43
Q

immunofluorescence microscopy can be used to detect specific _____ in fixed cells

A

proteins – have Ab with fluorescent dye attached

44
Q

immunofluorescence microscopy

A

Ab covalently linked to fluorochrome like Texas red or green fluorescent protein

when bound to antigen, lights up

45
Q

reporter gene

A

glow tells us where gene is active

46
Q

lower E = ______ wavelength

A

longer

47
Q

goat anti-rabbit

A

indirect immunofluorescence

immunize a goat with Fc common to all rabbits to get better fluorescence upon binding

48
Q

epitope tag

A

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
Q

2 problems with fluorescent microscopy

A

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
Q

zebrafish microscopy

A

have clear embryo

fluorescence tagging used to see what grows where in growing embryo

51
Q

deconvolution microscopy

A

collect into overnight and computer takes out blurry info

52
Q

confocal microscopy

A

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
Q

focal plane

A

sliver of a thick specimen

54
Q

laser scanning confocal microscopy (LSCM)

A

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
Q

total internal reflection fluorescence (TIRF)

A

visualize a thin focal plane like kinetics of microtubules and actin filaments

56
Q

fluorescence resonance energy transfer (FRET)

A

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
Q

super resolution microscopy is a subset of ________ microscopy

A

fluorescence

58
Q

photo activated localization microscopy (PALM)

A

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
Q

strategy x

A

slightly different times of excitation and release

60
Q

electron microscopy

A

electromagnetic lens focuses a high-velocity electron beam

living material cannot be visualized

61
Q

transmission electron microscopy

A

visualize things that have been negatively stained with heavy metal

62
Q

cryoelectron microscopy

A

view biological specimens using TEM if the specimen is frozen

ex. virus capsids

63
Q

scanning electron microscopy (SEM)

A

view surfaces of unsectioned metal-coated specimens

limited by thickness of metal coat

Cellular and Molecular Biology (34 decks)

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