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64 Cards in this Set
- Front
- Back
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What are 3 ways you can increase resolution (decrease D) |
1. Move lens closer by switching to a higher power 2. Place a single drop of immersion oil which has a higher N than air. 3. Decrease lambda used. Blue light is shortest. 450nm |
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What is the equation for the limit of resolution? |
Do on paper |
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Magnification = |
size of image _______________ actual size of object |
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Does it have units? |
No |
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Label a compound light mircoscope |
on paper |
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Describe the field iris diaphragm |
Located at base and controls diameter of visible illuminated field |
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The scanning objective lens is also the? |
Least powerful, 3.5 or 5 x lens |
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Describe the ocular lenses and how to focus them. |
They further magnify by 10X. 1. Pull apart and slowly close while looking through both lenses until you see a single image. 2. Focus one ocular using fine/coarse knobs 3. While looking through both, rotate the other lens barrel until it to is focused. |
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What are the 3 main parts to a Koehler illumination? |
1. Focus specimen on 10X and close condenser diaphragm completely 2. Get edges of small circle of light as sharp as possible by moving the condenser unit, usually into it's highest position. Center Image 3. Open FID and adjust condenser diaphragm for comfortable light. |
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Pro Tip: When switching from lower to high magnifications, you must _____________ the FID and __________ the condenser dia, b/c the higher power lenses are less efficient at gathering light. Simply turning up the brightness will ______________ the resolution |
close down open reduce |
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Higher power lenses have a _________ depth of focus |
shorter |
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Higher power lenses have a _________working distance |
shorter |
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Higher power lenses have a _________field diameter |
smaller |
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For calibrating an ocular micrometer, how long is the scale of the stage micrometer? |
1.0 mm |
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The stage micrometer is the ruler _________ numbers engraved on it? |
without any |
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We use _____ type pipettes |
TD, to deliver |
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A ______ TD pipette is emptied by having the total volume blown out |
blowout |
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A ________ TD pipette is emptied with still some solution in tip |
gravity |
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Draw out the scale for autopipettor measurements |
brah |
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What is an autoclave? |
A high pressure steam sterilization |
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What are two advantages of cryopreservation? |
1. Ensure a living back up of cells to ensure their availability in the future 2. Allows for many identical replication of an experiment using identical cells |
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During the cooling process, ice crystals form outside the cell, effectively _________ the solute concentration outside the cell causing the cell to ___________ from dehydration
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raising crenate |
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Are CPAs hydrophilic or hydrophobic? |
hydrophilic |
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How do CPAs prevent dehydration, salt toxicity and ice crystal formations? |
They decrease the freezing point of the solution by binding water molecules colligatively |
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For best results cell cultures are grown to _______ phase ? |
log |
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Why should cells be placed on ice ? |
to reduce the toxic effects of CPAs |
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How are CPAs washed off during thawing? |
centrifuged slowly, discard supernatant and replace with fresh media |
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A methylene blue-negative cell is one that is _________ in color |
white or colorless |
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Why? |
B/c metabolism oxidizes MB into a colorless comp. |
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Why is blood a special type of connective tissue? |
Because it's ECM is fluid |
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Differential cent. can produce which 3 protein containing fractions? |
Plasma lysate and membrane fractions |
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What are the main componentss of plasma frac? |
serum albumin, globulins, clotting and hormorne-binding proteins |
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What are the main components of lysate frac? |
protein hemoglobin from erythrocyte cytoplasm |
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What are the main components of membrane frac? |
integral membrane proteins, glycoproteinsm and cytoskeleton proteins |
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What are safety considerations of Beta-mercaptoethanol? |
respitory tract, eye and skin burns |
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What are some causes during a fractionation procedure that result in losses of materials? |
incomplete seperation of its components, materials sticking to surfaces. make a compact pellet good |
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In a Bradford assay the acidified components of coomassie brilliant blue bind quantitatively to ______ and __________ residues in the proteins |
arginine and aromatic AA (tryosine, phenylalanine and tryptophan) |
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Which causes a shift from _____ to ____ |
470 to 595 |
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Safety considerations of Acrylamide |
neutoxin, carcinogen, mutagen and reproductive toxin |
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What is a tracking dye often used in electrophoresis? |
Bromophenol blue |
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SDS is used to bind to proteins to create an overall negative charge. When the charge to mass ratio is the same the proteins will migrate due to the molecular sieving properties |
notes |
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SDS is used to _________ proteins to the same linear shape so that they are separated equally based on molecular weight |
denature |
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Beta-mercaptoethanol does what to proteins? |
It reduces disulphide linkages |
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What effect does this have b/w proteins and SDS |
It allows proteins to bind to SDS and assume a random coil formation which will separate based on weight |
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What does Western blotting, a method of visualization utilize? |
antibodies to detect certain proteins |
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What is Tetrahymena? |
a freshwater, unicellular, ciliated protzoan that has many attributes that make it ideal for studying |
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Describe two reasons why Tetreahymean is a model organism. hint: obtain and size |
1. It is easily obtained from fresh water ponds and can be grown in a simple culture media 2. It is large enough to be seen under a microscope, so it is suitable for studying ciliary motion, organelle structure and function, and behavior |
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What is a kinetie? |
a longitudinal tract of arranged cilila |
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What is the composition of a cilia |
a '9+2' arrangement of microtubules |
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What is the monomer of microtubules? |
tubulin |
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_________ is a side effect of the ciliary motion which causes Tetrahymena to spin as they move and it helps them phagocytise by propeling suspending food particles towards its _________ grove |
The vortices oral |
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In phagocytosis food particles are enclosed in membrane bound food vacuoles, or ______________ |
phagosomes |
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Each phagosomes will fuse with a lysosome to form a _______________ |
phagolysosome |
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Following digestion where are released nutrients diffuse or are transported into the ______? |
cytosol |
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Food vacuole formation is a _______dependant process |
microfilament |
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What is monomer of microfilament? |
alpha-actin |
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The microfilaments which make up the cytoskeleton undergo cycles or polymerization and ________________ |
depolymerization |
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What are 3 main areas where microfilaments of the cytoskeleton are essential? |
1.vacuole formation 2. cleavage furrow during mitosis 3. actin in muscle contraction |
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What are the effects of Cytochalasin B? |
It inhibits polymerization of actin into microfilaments |
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What is India ink |
An aqueous suspension of fine carbon particles |
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Tetrahymena are grown in proteose peptone, which is a partial ___________ . What is its role? |
digest of animal protein. It provides soluble nutrients for the cells as well as supporting the growth of bacteria which Tetrahymena normally feeds on |
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What is sample error?
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Where an individual sample varies randomly from the whole population
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How can sample error be reduced? |
By sampling as soon after vortex for example. This ensures that the population is reasonably represented |
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What are 3 examples of sources of human error? |
1. limited precision of measuring device. 2. degradation of cellular components at room temperature 3. incomplete separation of constituents such as the pellet and the supernatant. e.g. leaving behind a small amount of supernatant. |
