Module 1

Question 1

fluorescent antibodies are typically used on

Answer 1

single cell layers in a. culture dish

Question 1

immunofluorescence

Answer 1

a process by which scientists can view which cells express a protein of interest using a antibody with a fluorescent tag using a fluorescent microscope
- allows viewing of even sub cellular components

Question 2

immunocytochemistry

Answer 2

• viewing a protein of interest within layer of cells

Question 3

immunohistochmeistty

Answer 3

visualizing that protein within a tissue section

Question 4

what is common between immunoxcytocehmsitry and immunohistochemistry

Answer 4

both involve detection of proteins with anitobdies

Question 5

histology

Answer 5

ability to recognize teh normal structure of issues
- allows understanding of when things go wrong in pathologies

Question 6

how do histological studies work (process of obtaining tissue)

Answer 6

• preserve tissue
• embed tissue in wax
• stain tissue
• microscope

Question 7

what is the importance of preserving tissue in histology

Answer 7

keep it in a lifelike state and prevent degradation

Question 8

why is it important to embed the tissue in wax for immunohistochemistry

Answer 8

• provide stability to be bable to cut thins sections without damage of the tissue
• and thin sections are needed to allow light o apass through the section so it can be viewed

Question 9

what is it important to stain tissues in histology

Answer 9

• no innate colour, different dyes are used, otherwise the tissues are too transparent to see

Question 10

alizarin red

Answer 10

stains calcium red

Question 11

cresyl violet

Answer 11

stains neruons and glia purple

Question 12

alcian blue

Answer 12

stains muffins and sulphated glycoproteins blue

Question 13

haemotoxylin

Answer 13

stains nuclei blue

Question 14

eosin

Answer 14

stains cytoplasm pink

Question 15

oil red O

Answer 15

stains lipid droplets red

Question 16

micro-siruis red

Answer 16

stains collagen fibres 1 and 3 red

Question 17

immunohistochemistry process

Answer 17

• removal of wax
• rehydration of tissues
• antigen retrieval
• add h2o2 incubation (blocking solution)
• incubation of sample with secondary antibody
• colourimetric reaction is used to view protein of intrerset

Question 18

why must you remove wax in immunohistochemistry

Answer 18

to analyze the tissue

Question 19

why do tissues have to be dehydrated in immunohistochemistry

Answer 19

because tissues are dehydrated prior to embedding

Question 20

antigen retrieval purpose in immunohistochemistry

Answer 20

tissue was fixed in aldehyde containing solution; these aldehydes can mask the protein epitope that would be recognized by the antibody

Question 21

incubation with H2O2 in immunohistochemistry

Answer 21

• used when final antibody has an enzymatic tag like peroxidase
• the proteins in the blocking solution prevent the primary antibody from binding to regions of the gas slides.

Question 23

what is co-localization

Answer 23

• addition of more than one antibody to visualize two or more proteins I a cell and their locations in that cell

Question 24

what are the two ways to prepare the tissues before sectioning them into thin slices

Answer 24

- embedding in paraffin wax - quick freeze procedure + cyrossecitoning (sectioning) ** choosing between them deepens on the type of tissue or antibody that will be used for subsequent analysis

Question 25

how is water replaced in dehydrated tissues in immunohistochemistry

Answer 25

alcohol with increasing concentrations of water

Question 26

in immunohitochemistry, how is the wx dissolved after embedding