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ANHB3323 > 4: 4 > Flashcards

4: 4 Flashcards

1
Q

name from soft to hard tissue types best found in each one

A

fat > brain >muscle >cartilage > tendon > cortical bone

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2
Q

what gel works for fat + mammary cells

A

agarose

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3
Q

what gel works for brain + muscle

A

PA gel

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4
Q

what gel works for muscle + cartilage

A

PDMS

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5
Q

what gel works for cortical bone

A

glass

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6
Q

what are gold nanonrods used for

A
  • Digital stiffness writing

Gold nanorods in hydrogel // when hit with laser in specific pattern= creates heat

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7
Q

what is micro contact printing

A
  • Microcontact printing= size/shape variables + which one best works in stiffness
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8
Q

describe elctrospinning

A

ECM fibre arrangement random // spinning replicates

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9
Q

What can be used to measure ECM properties

A

AFM

micro indentation

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10
Q

what can be used to measure traction force

A

TFM
Micropillar
FRET

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11
Q

What is the wettablity test

A

Hydrophilicity or Hydrophobicity

o Sdd water + see how much contact angle - larger the angle= more hydrophobicity

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12
Q

what can you use to measure topography

A

Electro microscope= SEM

or AFM

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13
Q

what tests are used to measure the mechanical properties of ECM

A

A. Compression test: how much force it requires to x%
B. Tensile test: extend to x% (can be till rupture)
C. Rheological force test: how much force wants to go back to original state after rotation

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14
Q

how do Micro indentation test + atomic force micrsopce work generally

A
  • Indent arm/tip goes down in given force on a sample > detects changes in reflection when tip bend (soft sample = longtime till bend)
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15
Q

what’s the differences between Micro-indentation test and AFM

A

Micro= larger probe = whole cell tested

AFM= smaller probe // one integrin binding type test

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16
Q

what can AFM measure

A

force, stiffness, rupture, intracellular traction, height, adhesion

17
Q

how does AFM measure adhesion

A

when tip comes back up= adhesion/sticky clings to it = greater peak towards neg = more sticky)

18
Q

how does AFM measure height/topogrpahy

A

(as you know where you started (lateral height- length from starting point)

19
Q

On a graph for AFM is the steady rising line softer or harder compared to the quick up change

A

steady= softer

20
Q

how does AFM measure rupture

A

Coats AFM tip with integrin= binds to extracellular matrix > pull to see how integrin interacts with substrate i.e. stiffness
o = to rupture points
o Stiff= high force, low height - soft= low force, high height

21
Q

how does AFM measure intracellular traction force

A

uses antibody - protein interaction
o pulling force from cells
o Soft= little stable focal adhesion> little spread + low integrin = little traction force

22
Q

what does direct cellular measurement measure and what are the two types

A
  • Measures Traction Force
  • Types:
    1. Micro-Pillar
    2. Traction Force Microscopy
23
Q

pros/negs of Micro-Pillar

vs Traction Force Microscopy

A

Pillar pro= high res, no FEM beads, cons= no z

TFM pro= xyz data, cons= require confocal, FEM

24
Q

what is microvillar made from and how to variable it

A
  • Microneedle = made from PDSMs

- Variables: stiffness of PDMS, diameter, length

25
Q

what happens if you increase the height of a micropillar

A
  • Length Variable: displacement more on top of longer with same force as smaller (which doesn’t move a lot in comparison)
    o i.e. shorter force required in long
26
Q

why optimal aspect ratio exist for micropillar

A

too high= lateral collapse , too small = sagging

27
Q

how is TFM measured

A

2 Image Process:
Image 1: stressed configuration- cell exerting traction toward centre // beads following it inside
Image 2: remove cell (using trypsin) + beads go back
- Measure bead displacement between stressed and unstressed configuration

28
Q

what is FRET

A
  • Fluorescence Resonance Energy Transfer (FRET)

o Used to determine how “close” to fluorophores are

29
Q

how does FRET work

A
  1. Energy/laser directed at sample (green) > hits (excites+ changes energy stage) > energy released/emission (emits at slightly diff wavelength
  2. Donor Emission light can excite acceptor (if they are in close proximity)
  3. // can measure distance
30
Q

what is an example of something FRET can measure

A
  • Can Measure: Change in conformation= change in distance

o i.e. when fibronectin is pulled apart (Talin/vinculin to attach)

31
Q

design experiment to identify the effect of stiffness on traction force

A

. stiffness hydrogel + use beads to measure traction force using TFN

32
Q

Advantages of human on a chip

A

Pros=
- low cost alternatives as uses low fluid/med delivery
- mimic complex structures
0 increase success rate + speed of drug development

33
Q

limitations of human on a chip

A
  • not yet engineered intergrated systems
  • finding common cell medium to integrate system hard
  • ## limited self-renewal potential
34
Q

What is one success of lab on a chip

A

Microstructure - cancer metastasis
• Found that cancer cell can reassemble cancer nucleus after fitting through small holes
o // differentiate between healthy/tumor cells by how fast they move thru holes

ANHB3323 (14 decks)

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