Microscope

Question 1

What is Chromogenic Staining

Answer 1

Stains nuclear DNA purple (haematoxylin) and Cytoplasmic Proteins pink (Eosin)

Question 2

What is DIC

Answer 2

Differential interference contract. Is a light microscopy with 2 light beams, is related to refractive index. Shows major organelles. For thick and to observe small details

Question 3

What is Phase contrast

Answer 3

Refractive and un-refracted light, dark outline. for Thin cells

Question 4

What to use on moving molecules

Answer 4

Use a time-scale

Question 5

Steps for dead specimens

Answer 5

First fix the cell, embed in wax, slice and view under microscope

Question 6

Types of light microscopy

Answer 6

Phase-Contrast and Nomarski and bright field

Question 7

What is FACs

Answer 7

fluorescence- activated cell sorter

Question 8

Flow cytometry

Answer 8

fluorescence activated cell sorter isolates specific cell types, can use FACs to seperate specific cells

Question 9

Example of FACs sorting

Answer 9

Sorting T cells: T cells fluoresce both green and Red as as a tagged antibody specific for certain proteins is added they have THY 1.2 ad CD3

Question 10

Differential Centrifugation

Answer 10

Based on mass, spins and heaviest goes to bottom

Question 11

What is immunofluorescence

Answer 11

detects proteins using fluorescence tagged antibodies direct and indirect.

Question 12

Confocal Microscopy

Answer 12

Does one plane therefore allows to not have blurred images

Question 13

fluorescence microscopy

Answer 13

Can view specific molecules in live cells e.g. using stains like eosin an hematoxylin

Question 14

When same mass what can you use

Answer 14

density gradient

Question 15

What can you do if you can’t get a specific antibody

Answer 15

You can epitope tag the c or n terminus

Question 16

What can epitope tagging an organelle show

Answer 16

Show if it is in the cytoplasm or inside an organelle

Question 17

How to tell the orientation

Answer 17

C or N terminus and have a tag which will fluoresce when attaches and use a detergent digiton or Triton-X, can see if in organelle or just in cytoplasm

Question 18

What is TEM

Answer 18

Transmission electron microscope

Question 19

What does TEM do

Answer 19

Detects differences in density, high resolution. Focus on high electron densities rather then florescence. Electrons emitted from a filament and accelerated in an electric field. Then condenser lens focuses on, then scanning and objective specimen an dthen detector

Question 20

What is SEM

Answer 20

Scanning electron microscope

Question 21

What does SEM do

Answer 21

metal-coated speciemen, in a high vaccum, allows veiwing of surface features

Question 22

What do they do to cells and tissues for EM

Answer 22

Cut into thin sections

Question 23

What is Immunoelectron Microscopy

Answer 23

Use an antibody with a heavy metal tag to veiw proteins

Question 24

Viewing shape with EM

Answer 24

Can stain sample with heavy metal to view shape