Microscopy

Question 1

What is microscopy?

Answer 1

using microscopes to view objects/specimens that are not visible to the naked eye

Question 2

What are the different components you need for microscopy?

Answer 2

• Light source
• Light conditioning system - optimise the light source to give best image
• Specimen - glass side, covered by embedding medium
• Objective - magnifying glasses
• Detector - way to detect the image

Question 3

What is the use of a incubator box?

Answer 3

used for the specimen combined with air heater to ensure the temperature or level of carbon dioxide remains equilibrated and tightly controlled

Question 4

What is light microscopy?

Answer 4

• Light to illuminate
• Lenses to magnify
• By increasing the contrast, allows to see more detail of the cell
  
  • DIC
  • Phase
  • Colour brightfield

Used to see histology of tissue

• Identify areas of issue
• But don’t see how the areas interact with each other

Identify general patterns of distribution of proteins of interest

Shows cell morphology - eg. Intact collagen and denatured collagen

Can be used to see dynamic images in time-lapse

Question 5

What is electron microscopy?

Answer 5

• can either be transmission or scanning

Question 6

What is fluorescence microscopy?

Answer 6

Uses a laser beam and filter cube to give image

Stokes shift = due to energy loss the emitted light is shifted to longer wavelength relative to the excitation light (2 on graph)

Question 7

What is photobleaching?

Answer 7

Bleaching of flurochromes

• due to high intensity illumination the fluorophores might loose permanently their ability to emit light