Cellular Physiology Flashcards
Name the different stages of the cell cycle?
G0-resting phase G1 - growth phase, increase volume S - synthesis phase, synthesis of DNA G2 - growth phase, ready for division Mitosis
What does PCNA stand for and what is it?
Proliferating cell nuclear antigen.
Takes part in interactions between DNA and DNA-polymerase during replication. Used as a common proliferation marker.
Where is PCNA located and in which part of the cell cycle does it reach the highes level?
Nucleus
S - phase
Name the two most common proliferation markers?
PCNA
Ki-67
What is the detection method called for proliferation of cells?
Immunohistochemistry
using histological sections.
What is the detection by visualization of PCNA based on?
Specific antigen-antibody binding and antibody visualization.
What do the PCNA method apply for?
Fluorecent labelling, conventional light microscopy, long term preservation of sample.
Describe the multiple-step procedure of PCNA?
- antigen-antibody reaction
- primary antibody-secondary antibody reaction
- bioton-enzyme binding (POD)
- enzyme-substrate reaction
- counterstaining (hematoxylin)
What is used for deparrafinization?
xylen
what is used for rehydration?
ethanol series in decreasing order (100%, 96%, 70%, 50%, distilled water)
What is used for washing?
PBS (Phosphate buffered Saline)
What is chromogenic substrate?
DAB (diaminobenzidine)
gives brown colour
What is used for dehydration?
Increasing series of ethanol.
What are immunohistochemical reactions based on?
compatibility of an antigen and the corresponding antibody.
What is an antigen?
Antigen –> Antibody generating, any substance able to evoke an immune response (antibody production)
How can antibodies be produced?
Injection of antigen into an animal followed by antibody separation from the serum.
Preparation in hybridomas (immortal antibody producing cells), cultures –> collection.
What is proliferation activity rate?
percentage of the proliferating cells from the entire number of cells within the cell population of interest.
What is proliferation rate evaluation an important part of?
related to renewal and cell cycle. Important tumour tissue examination method.
What is the name of the place that binds the antigen?
FAB-fragment
Which cells can be found in the small intestine?
Enterocytes, Goblet cells, Stem cells, Paneth cells, M-cells, Enteroendocrine cells.
