- What are the components of cell culture environment?
o media and media components
o the plastic the cells are attached to
o the gas composition
- What are the components of basal cell culture medium? What components you do not expect to see in it?
Basal medium contents include water soluble vitamins, trace elements, amino acids. It lacks serum and antibiotics
- Do we always add antibiotics to medium?
No, only when necessary
- What is the purpose of complete medium?
To support growth and proliferation of animal cells
- What is the purpose of sodium bicarbonate buffer?
To help maintain the correct pH when CO2 is present
- Is atmosphere inside a biological safety cabinet absolutely sterile?
It is very clean but not absolutely sterile
- What is the purpose of adding serum into media?
o To neutralize toxic nutrients, trypsin and other proteases
o Source of growth and adhesion factors, hormones, lipids and minerals for the culture of cells in basal media
o Regulates cell membrane permeability and serves as a carrier for lipids, enzymes, micronutrients, and trace elements into the cell
How cell surface proteins that promote cell-cell contact will affect cells in culture?
Cells stop growing due to contact inhibition
- Confluency refers to:
The ratio of area occupied by the cells to the total area available
- Why cell growth schedule analysis is important?
o Allows to establish a cell doubling time
o Allow to see changes in growth patterns over time
o They give record of growth
- During the growth of animal cells it is important to keep cells in which phase of growth curve?
Log phase
- What does PBS stand for?
Phosphate buffered saline
- The purpose of a disassociation medium (trypsin) is to:
To cause adherent cells to detach from one another and the substratum
- The purpose of a medium containing FBS after trypsinization is to
Reduce further enzymatic activity
Phalloidin selectively binds F-actin. What is F-actin?
Filamentous actin
Phalloidin is a poison which affects the depolymerization of actin. A cell treated with phalloidin would likely exhibit difficulties with motility via pseudopodia. True or false?
True
. Which statement about Rhodamine-Phalloidin staining is NOT TRUE
A. Optimal for fixed and permeabilized samples
B. Very widely cited fluorescent phalloidin conjugate
C. Only used for suspension cell cultures
D. Superior to antibody staining
C. Only used for suspension cell cultures
. What dye is used in the rhodamine phallodin staining?
TRITC
Is the excitation wavelength more or less than emission wavelength?
Less than the emission wavelength
What regions of DNA will Hoechst fluorescent dye most likely bind?
A-T regions of DNA
What kind of microscope was used to visualize result in the cytoskeleton staining lab experiment?
Fluorescent
What are the steps of immunochemistry?
Cell seeding
Immunostaining
Imaging
Image analysis
What is the purpose of using FBS in immunocytochemistry protocol?
To block the cells
. Name here types of ICC reporters
o Enzyme-coupled antibody
o Fluorophore-labeled antibody
o Biospecific small molecule dye
