Cellular Physiology

Question 1

Name the different stages of the cell cycle?

Answer 1

G0-resting phase
G1 - growth phase, increase volume
S - synthesis phase, synthesis of DNA
G2 - growth phase, ready for division
Mitosis

Question 2

What does PCNA stand for and what is it?

Answer 2

Proliferating cell nuclear antigen.

Takes part in interactions between DNA and DNA-polymerase during replication. Used as a common proliferation marker.

Question 3

Where is PCNA located and in which part of the cell cycle does it reach the highes level?

Answer 3

Nucleus

S - phase

Question 4

Name the two most common proliferation markers?

Answer 4

PCNA

Ki-67

Question 5

What is the detection method called for proliferation of cells?

Answer 5

Immunohistochemistry

using histological sections.

Question 6

What is the detection by visualization of PCNA based on?

Answer 6

Specific antigen-antibody binding and antibody visualization.

Question 7

What do the PCNA method apply for?

Answer 7

Fluorecent labelling, conventional light microscopy, long term preservation of sample.

Question 8

Describe the multiple-step procedure of PCNA?

Answer 8

1. antigen-antibody reaction
2. primary antibody-secondary antibody reaction
3. bioton-enzyme binding (POD)
4. enzyme-substrate reaction
5. counterstaining (hematoxylin)

Question 9

What is used for deparrafinization?

Answer 9

xylen

Question 10

what is used for rehydration?

Answer 10

ethanol series in decreasing order (100%, 96%, 70%, 50%, distilled water)

Question 11

What is used for washing?

Answer 11

PBS (Phosphate buffered Saline)

Question 12

What is chromogenic substrate?

Answer 12

DAB (diaminobenzidine)

gives brown colour

Question 13

What is used for dehydration?

Answer 13

Increasing series of ethanol.

Question 14

What are immunohistochemical reactions based on?

Answer 14

compatibility of an antigen and the corresponding antibody.

Question 15

What is an antigen?

Answer 15

Antigen –> Antibody generating, any substance able to evoke an immune response (antibody production)

Question 16

How can antibodies be produced?

Answer 16

Injection of antigen into an animal followed by antibody separation from the serum.
Preparation in hybridomas (immortal antibody producing cells), cultures –> collection.

Question 17

What is proliferation activity rate?

Answer 17

percentage of the proliferating cells from the entire number of cells within the cell population of interest.

Question 18

What is proliferation rate evaluation an important part of?

Answer 18

related to renewal and cell cycle. Important tumour tissue examination method.

Question 19

What is the name of the place that binds the antigen?

Answer 19

FAB-fragment

Question 20

Which cells can be found in the small intestine?

Answer 20

Enterocytes, Goblet cells, Stem cells, Paneth cells, M-cells, Enteroendocrine cells.