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Body logistics > BL10 L2 Examining Cells And Microscopy > Flashcards

BL10 L2 Examining Cells And Microscopy Flashcards

1
Q

List the 4 types of tissues

A
  • striated muscle
  • nerves
  • connective
  • epithelial
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2
Q

Resolution definition

A

Smallest distance that two objects can be separated and seen as two different objects

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3
Q

Mili

A

1mm = 1x10^-3m

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4
Q

Micro

A

1um = 1x10^-6m

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5
Q

Nano

A

1nm = 1x10^-9

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6
Q

List 5 biopsy techniques

A
  • surgery and later dissections
  • scraping methods e.g. scalpel, scrapes, curettes
  • sharp needles e.g. needle/ punch biopsy, pipelle
  • direct venepuncture e.g for blood smear
  • tranvascular: device travels through BVs to site of biopsy e.g. liver, lungs, brain, heart
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7
Q

Examples of scaring biopsy methods

A

Curettes
Scalpel scrapes

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8
Q

Examples of sharp needle biopsy techniques

A

Needle biopsy
Punch biopsy
Pipelle

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9
Q

What can a pipelle be used for?

A

Endometrial biopsy sample
Endometrium-Uterus lining

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10
Q

What does transvascular mean and what can it be used to take a biopsy of?

A

Device travels through BVs to biopsy site
Kidney, lungs, brain, heart

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11
Q

2 reasons for fixation

A
  • samples need to be translucent + thin
  • protect against microbes
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12
Q

How thin do samples need to be for fixation

A

2-10 micrometres

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13
Q

Why do samples need to be translucent and thin?

A

To avoid diffraction of light and prevent blurry image

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14
Q

Why do samples need to be fixated to protect against microbes?

A

Once of out the body, samples are no longer under the protection of the immune system

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15
Q

What fixative is used?

A

Formalin solution - 37% solution of formaldehyde

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16
Q

How long do you fixate for?

A

24-48 hours only

17
Q

What happens if you fixate for too long?

A

Shrinkage occurs due to dehydration
Causes fixation artefacts

18
Q

What causes fixation artefacts?

A

Fixation for longer than 48 hours
Causes shrinkage due to dehydration

19
Q

How does paraffin wax embedding occurs?

A
  • dehydrating in varying alcohol cones
  • immersed in hot paraffin wax overnight
  • tissue orientated in mould
  • add more wax
  • allow to solidify
  • slice thinly using microtome
20
Q

What machine is used to slice samples thinly?

A

Microtome

21
Q

Frozen section process

A
  • specimen frozen rapidly
  • frozen specimen cut with microtone
  • stained with H+E
22
Q

Difference between paraffin embedded tissue section and frozen sample

A

Paraffin - fixed tissue, 24-48 hours to make, permanent, clear/detailed
Frozen sample - fresh tissue (preserves lipid containing organelle), 10-20 mins to makes, lasts a few months, lacks detail

23
Q

What colour does haematoxylin stain and what does it stain?

A

Blue
Nucleus - binds to acidic things e.g DNA, RNA

24
Q

What colour does eosin stain and what does it stain?

A

Pink
Cytoplasm, extracllular matrix

25
Q

What colour does H+E stain and what difference does it make to only using one?

A

Purple
Nuclei stained clearer
Cytoplasm more detailed

26
Q

Tissue prep for light microscope

A
  • fix with formalin
  • embed with paraffin wax
  • stain e.g. H&E, methylene blue
27
Q

Tissue preparation for electron microscope

A
  • fix with glutaraldehyde
  • embed with epoxy resin
  • stain e.g. osmium tetroxide
28
Q

What has a high resolution, light or electron microscope?

A

ELECTRO MICROSCOPE
Light- 0.25 MICROmetres
Electron- 0.25 NANOmetres

29
Q

What has a higher magnification, light or electron microscope?

A

ELECTRON MICROSCOPE
Light- x600
Electron- x500,000

30
Q

What can be viewed with light microscopes compared to electron microscopes?

A

Light- living and moving
Electron- dead and inert (have to be vacuumed)

31
Q

Colours seen in light vs electron microscope

A

Light- natural colour
Electron- black and white

32
Q

Financial costs of light vs electron microscopy

A

Light- cheap and easy prep
Electron- expensive and difficult prep

33
Q

What type of image does SEM give?

A

3D
Black and white

34
Q

What image does TEM give?

A

2D
Black and white

35
Q

Advantage of cell culture

A
  • complete control over physical environment
  • homogeneity (the same) of sample
  • reduced need for animal models
36
Q

Disadvantages of cell culture

A
  • hard to maintain so skilled worker needed
  • high cost for small amount
  • 3D is lost
  • aneuploidy - abnormal number of chromosome in haploid organism
37
Q

What can be visualised in dark field and how does it work?

A
  • living cells
  • illuminates sample with light that won’t be collected by objective lens so wont form part of image
  • very dark background with bright objects on
38
Q

How does confocal microscopy work?

A
  • laser excites fluorescent dye
  • electrons raised to higher energy level
  • wavelength is emitted as electron returns to ground state
  • emitted light is sent through mirrors and pinhole screen to CMOS detector
39
Q

What can confocal microscopy be used for clinically?

A

Evaluation of various eye diseases

Body logistics (20 decks)

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