The ability to see two points that are close together as separate points
A statement or prediction of no difference
A writing by people who actually performed the experiments and are presenting their data
The differentiated cell in the HL-60 cell lineage that is most common in PMA treated cells
the process of an extracellular signal being converted into an intracellular one, to elicit to a specific cellular response
the transition of cells to a committed cell type, one which is more specialized than its precursor
the use of DNA template to form a complementary RNA sequence
the dye used in our lab to view morphological differences in differentiated HL-60 cells
an enzyme that catalyzes the cleavage/degradation of RNA
A membrane protein involved in cells binding to other cells or to the extracellular matrix (like fibronectin)
An enzyme that uses RNA as the template to make cDNA
the liquid material left above the pellet after centrifugation
an enzyme that uses DNA as the template to make new DNA strands
the device on a light microscope that regulates the amount of light that passes through the specimen
Cells that are maintained in culture have undergone ------------ so that they become immortal and can be maintained indefinately
What is NOT required for PCR?
PCR is a product of ?
The H in HL-60 cells stand for?
What is the central dogma of biology?
the transfer of genetic information from DNA --> RNA--> Protein
What is the equation for counting total cell volume?
(total of 4 squares) (10^4)
What is PCR an abbreviation for?
polymerase chain reaction
Why does treating cells with PMA increase their ability to bind to fibronectin coverslips?
PMA is colony stimulating factor. HL-60 cells will differentiate to monocytes which have integrins on their cell surface that will bind to fibronectin coverslips
Why is it useful for a microscope to be parfocal?
Then the subject will remain focused even though there is a change in magnification
Why is it important to keep isolated RNA sterile and on ice?
RNases degrade very easily, which is why it is important to keep a sterile environment
You want to observe the chlorophyll molecules within the chloroplast of a plant cell. What type of microscope would you use?
Transmission Electron Microscope
Purpose of matrix metalloproteases
to digest the extracellular matrix so they can move around
Colony Stimulating Factor
are secreted glycoproteins which bind to receptor proteins on the surfaces of hemopoietic stem cells and thereby activate intracellular signaling pathways which can cause the cells to proliferate and differentiate into a specific kind of blood cell (usually white blood cells)
simple, single-stranded oligonucleotides that are sequence specific (to a gene for example)
protein that binds to integrins and plays a role in cell surface adhesion
Distinguish between a pluripotent cell and a totipotent cell
pluripotent cells can only give rise to certain cells while totipotent cells can give rise to a new organism or part due to the ability to give rise to any cell
Name and Describe the three steps in PCR?
1. Denaturation of the cDNA - splits the strands 2. Annealing of foward and reverse primers - the primers bind to appropriate DNA 3. Elongation of primer to produce a product - polymerase enzyme adds nucleotides for elongation
Steps involved in the scientific method
observation, hypothesis, collect data, analyze results, final conclusion
What instrument would you use to accurately measure 75ml?
What instrument would you use to accurately measure 32ul?
What instrument would you use to accurately measure 3ml?
Sections in a journal article
abstract, introduction, materials and methods, results, conclusions
volume range of p20
2uL to 20uL
volume range of p200
20uL to 200uL
volume range of p1000
200uL to 1000uL
an undifferentiated precursor cell that can divide indefinately and gives rise to other tissue specific cells
process by which a less specialized cell becomes more specialized
Tumor Necrosis Factor
an inflammatory cytokine that is produced by white blood cells (monocytes/macrophages), a cytokine that causes inflammation, apoptosis, inhibit tumorigenesis, and inhibit viral replicaiton
a term used to describe protein molecules that are secreted by cells (especially in the immune system) that serve as chemical messengers or regulator molecules, a molecule that allow cells to communicate and alter one another's function
Name at least 8 parts of the microscope:
ocular lens, rotating head, nosepiece, objective lens, arm, base, stage, condenser, iris diaphragm, coarse adjustment, fine adjustment, mechanical stage control, illuminator, dimmer
What are the two types of electron microscopy? Which one has the highest magnification?
Scanning EM and Transmission EM. Transmission is the highest
Why do electron microscopes have higher resolution?
bc electrons are used to view the subject rather than just light. Electrons have a shorter wavelenth resulting in higher resolution
a special kind of slide that is commonly used to measure cell density within a culture
helps us see cell death
HL-60 cells are?
an enzyme that uses RNA as the template to make cDNA
Name two reasons why the HL-60 cell line is used as in in-vitro model system?
1. They are "immortal", so they will keep replicating as long as they are kept under the right conditions 2. They are pluripotent allowing them to differentiate into different types of cells
PMA stock solutions are prepared in ?
Electophoresis results is the seperation of molecules based on what two things?
size and charge
What are the four major groups of macromolecules?
proteins, nucleic acids, carbohydrates, lipids
uses a beam of electrons to scan the surface of an object
used to view very thin sections of a specimen rather than the surface
a process that begins with absorption of light by a molecule and ends with its emission
whatever is in the center of the field of view on low power will stay in the center as the power changes
2 Filters on a Flourescent Microscope
1. Exciter filter 2. Barrier Filter
the illuminating light passes through the exciter filter passing only the wavelengths they excite the fluorescent dye
blocks out light and passes only those wavelenghts when the dye flouresces
Substrates of MMP-9
collagen, gelatin, elastin, pro-TNF, and interleukins
Sodium Dodecyl Sulfate- denatures secondary and non-disulfide tertiary structures so the proteins lose their native shape
meaning that it contains both hydrophobic and hydrophilic regions...also is negatively charged
purpose of glycerol
to make the protein sample more dense than the buffer so it stays in the well of the gel
a buffer that allows you to follow the protein migration through the cell
detects the amount of enzymatic activity of a molecule or type of molecules present in a sample