Kiss intro to cell bio

Question 1

What are the 3 pros of confocal microscopy?

Answer 1

1. offers 3D rendering of cells
   2.live cell imaging (while cells are moving around)
   3.multiple snapshots per second

Question 2

How does confocal microscopy do 3D cell imaging?
What type of cell can we look at, what can we use to look at them?

Answer 2

-it uses a very precise laser that can focus on different focal planes to allow 3d imaging
-can look at live cells and can use markers to track their movement

Question 3

What are the 3 cons of confocal miscropscopy?

Answer 3

1. limitation of specificity of markers
   2.co localization only implies co function
   3.resolution is limited

Question 4

How are the specificity of markers limited?

Answer 4

-specificity of marker EEA1 is an early endosome
marker but this doesnt imply that other proteins that bind early endosome are present on that particular endosome

Question 5

why does co localization on imply imply co function, what is an example and dimension shows they arent together?

Answer 5

-Assumptions of what compartments we look at. Think the man and women shadow makes it look like they’re tg but the third dimension shows they are not tg. Thus just because they co- localize it does not mean they are tg

Question 6

How is confocal microscopy limited by resolution?
At what length does it look like the cells are together

Answer 6

-Resolution is limited by the wavelength of visible light
-350-700nm is bigger than the structures we are looking at which are 50nm, thus if 2 things are 50nm apart they look like they are tg or even if they are 300nm apart they look like they are tg but they could be far apart