Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

Building blocks of life > Methods in cell biology > Flashcards

Methods in cell biology Flashcards

1
Q

what is the issue of using cells from animal tissue or individuals?

A

they can vary from sample to sample and are genetically diverse lacking consistency

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

what happens to cells removed from their natural environment?

A

can become stressed and die without proper nutrients

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

how can you keep isolated tissue alive?

A

simple salt solutions should keep them for a few days

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

when do cultured cells die?

A

after a few generations ( 40-60)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

how do telomeres affect cell grwoth?

A

telomeres shorten following cell division and limit the number of times a cell a divide

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

how do tumour cells become immortal?

A

they can rebuild their temomeres so become immortal

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

where did HeLa cell orginiate?

A

henrietta Lacks 1952 cervial tumour

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

why did HeLa cells become a thing?

A

they were immortal and could be cultured indefinately

made into a cel line

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

how can we now make immortal cell lines?

A

infect them with SV40 which activated telomerase so its a normal cell with an immortal function

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

what do typical cell culture medias contain?

A 
salts 
amino acids
vitamins such as folic acid and riboflavin
glucose
re phenol dye
buffers
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

what does confluent mean?

A

cels run out of space in the dish and cover alll the space at the bottom
signals they need to be split into new dishes and replated

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

how are cells moved to a new dish?

A

an enzyme trypsin detaches them

they are passaged and allow to form new connections on the disk

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

are HeLa cells adhernent or not?

A

they are adherent

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

what does passaging mean?

A

its the process of allowing cells to make new attachements

needs to happen 1-2 times a week

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

what is a non-adhernt cell?

A

blood cells

require dilute medials in suspension

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

name some basic cell culture pricnciples

A

aseptic technique
kept in sterile conditions
no contimination

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

who made the first pimitive micrscope?

A

Robert Hooke 1665

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

what did the Van Leeuwenhoek microscope look like?

A

very advanced for the time

used to discover sperm, red bood cels, protists and muscle fibres

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

what are the three types of light microscope around today?

A

bright field
phase contrast
differential interference contract DIC

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
20
Q

what is H&E stain

A

most common staining method

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
21
Q

what does haemoatoxylin do?

A

stain acid strcutres such as nucleus

Purpleblue

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
22
Q

what does eosin do?

A

stain basic strctures such as cytoplasm

pink

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
23
Q

what colours does the azan trichrome stain use?

A

nuclei red
collagen/BM blue
muscule and RBC orange
used for connective tissues and epitheliums

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
24
Q

when were the stages of mitosis discovered?

A

1878 by Walher

25
Q

how was mitosis discovered?

A

dyes such as aniline used to visualise chromosomes

26
Q

what is aniline?

A

specialise DNA stain

27
Q

how do fluorescent microscopes work?

A

shine light at one wavelength at the cells and it reflects it at different wavelength
filters are used too

28
Q

how are antibbodies ued in fluorescent microscopes?

A

they can recognise a specific point or thing in the target molecule so you can see this certain thing
they bind to biological molecules
secondary antibodies can then bind with fluorescent markers for colours which amplify the signal

29
Q

how is the location of the fluorescence detected in the tissue?

A

microscopes

30
Q

why is live cell imaging used?

A

cells are dynamic and processes occur over time

want to visualise these processes rather than a snapshot of them

31
Q

how does live cell imaging work?

A

uses an inverted microscope

objective is placed under the micscope allowing view of cells in culture media

32
Q

how are cells kept alive in live cell images?

A

environment chamber maintains the temp, adjusts gas suply to maintain pH
kept alive and health for several days under the microscope

33
Q

what is GFP

A

a gene from a jellyfish

34
Q

what was the original form of GFP like?

A

worked best at sea temps and onyl weakly fluoreced, easyily bleached and unstable

35
Q

what is gfp like now?

A

mutant version workes at 37degrees and can now work as a monomer and is more photostable

36
Q

what are the uses of gfp?

A

track embryo development

protein dynamics

37
Q

what develoments are there for gfp now?

A

multi colours of it now can be used to track multiple things
eg brain neurones all together

38
Q

what is FRAP

A

fluorescence recovery after photobleaching

39
Q

when is FRAP used?

A

determine kinetics of protein redistrubution and detect how much of the proteinis immobile or membrane bound

40
Q

what other types of gfp are there?

A

RFP

isolated from coral

41
Q

what is intravital imagining?

A

the use of GFP in cells or tissues in live organisms being viewed

42
Q

what is photo activation

A

laser shined onto tissues and changed colour after stimulation
allows tracking of sub-population cells

43
Q

what arethe main limitations of photoactivation?

A
  • photobleaching
  • phototoxicity
  • overexpression
  • protein folding
  • cellular distrubution
44
Q

what is photobleaching?

A

limitation of photoactivation

fluorescent proteins are sensitive to bleaching so long term studies can be tricky

45
Q

what is photo-toxicity

A

limitation of photoactivation

illumination of cells with high-intensity light can be toxic formin free radicals

46
Q

what is overexpression?

A

limitation of photoactivation
tagged genes are often expressed at high levels to make a good signal
excess proteins can after the cells normal distrubution

47
Q

what is cellular distribution in relation to photoactivation?

A

the large fluorescent proteins may affect the cellular disrubution of the tagged proteins which is especially key for membrane proteins

48
Q

what is a confocal microscope?

A

modified version of fluorescent miscopscopy

49
Q

how does a confocal microscope work?

A

lasers of specific wavelength are used to illuminate molecules in the sample
3D image of the cell with cross sections

50
Q

advantages of a confocal microscope

A
  • 3D view
  • cross sections
  • info on spatial relationship
  • used on both cells and tissues
51
Q

advantages of TEM

A

highest resolution possible

52
Q

how does TEM work?

A

antibodies are labelled with gold to detect cellular structures

53
Q

the disadvantage of TEM

A

requires very thin sections of cells

54
Q

what is super resolution microscopy?

A

provides 3D view of cells within a tissue and looks at spatial relationships within the tissue

55
Q

can super resolution or confocal do timelapses?

A

no

laser light is scattered by skin and other tissues and does not penetrate much

56
Q

what is STORM?

A

stochastic optical resolution

57
Q

what does STORM do?

A

allows you to determines the exact centre of the fluorescent signal and therefore the exact location o the dye molecule

58
Q

limitations to STORM

A

can be very slow
photobleaching and toxicity can be an issue
limited use of GFP
expensive

Building blocks of life (17 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions