Kherallah 1

Bio Lab Major Quiz 2 Study Guide

I. Spearman Rank Correlation: H0 = no significant correlation between variables
a. Test statistic = rs (Spearman rank correlation coefficient)
i. Perfect positive correlation = +1.0
ii. No relationship = 0
iii. Perfect negative correlation = -1.0
b. Finding rs
i. Rank each variable group separately
ii. Find difference (d) for each pair of variables, square each d, then find sum
6 d 2
iii. rs  1 
n(n 2  1)
iv. Compare rs with tabular statistic in given table with different alpha’s
1. If absolute value of rs is less than tabular, cannot reject H0
a. You will lose marks if you say “accept H0”
2. If it’s greater, then reject H0
II. Chapter 5 – Microscopy and Microbiology
a. Need to know diagram of compound microscope on p. 58
b. w/ dissecting microscope, object is seen by reflected light
i. w/ compound microscope, light illuminated below, thus transmitted light
c. Substage condenser is lens focusing light on specimen
d. Color of object depends on wavelength that object does not absorb
i. i.e. wavelength transmitted or reflected
ii. Most cellular parts transparent – staining used (see Gram Staining below)
e. Light passing through objective is inverted
i. Eyepiece magnifies inverted image to make virtual image
1.  light only appears to be from points where image located
f. Resolution = ability of lens to make distinct images of 2 closely adjacent points
i. Smaller the distance, better the resolution
g. Resolving power = minimal distance separating two points that are distinguishable
i. Resolution inversely proportional to resolving power
ii. R.P. = 0.6λ/numerical aperture
1. Numerical aperture relates to how widely the light spreads from sample
a. Inscribed on barrel of objective
2. Thus R.P. improved by
a. Decreasing wavelength
b. Increasing numerical aperture
i. i.e. oil helps the numerical aperture by focusing more
ii. wider objective also helps
iii. Limitation of light microscopy
1. Human eye limits wavelengths used (can’t see out of vis spec)
2. Thus can’t improve R.P. any further than best wavelength
h. Magnification = ocular by objective (we use 10x)
i. Electron microscopes
i. e- are much shorter wavelengths
ii. Image received onto a device for adjustment and capture
iii. Transmission electron microscopy (TEM)
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1. Depends on transmission of e-
2. Requires extremely thin specimens
3. 2D image obtained
4. Specimens stained with heavy metals
5. Benefits = magnification/resolving power (molecules can be seen)
iv. Scanning electron microscopy (SEM)
1. Sample coated with platinum or gold etc.
2. Electrons excited by beam of electrons
3. Machine measures angles of electron scatter to form 3D image
j. Comparing microscopy types (copied from bio lab binder)

Type Min. Resolving Max Effective Appropriate

power magnification specimen thickness
Light 200-300 nm 1200x 1.5-15 µm
TEM 0.2-0.5 nm 1000000x 5-100 nm
SEM 5-20 nm 200000x Not critical
k. Gram Staining
i. Smear
1. Smear bacteria
ii. Heat-fix
1. To glue the bacteria to slide
iii. Stain
1. 1° stain = crystal violet
a. Only bacterial walls with enough peptidoglycan retain this
complex
2. Trapping agent = gram’s iodine
a. Complexes with CV to retain it in place on cell wall
3. Decolorization = alcohol/acetone
a. Gets rid of excess color
4. Counterstain = safronin
a. Gives the pink color in all the cell walls
5. Gram + = dark cells with peptidoglycan, Gram - = pink with not much
peptidoglycan in cells
III. Chapter 6 – Biochemical Pathways
a. Can be done by using mutant strains, each containing a different inhibition in pathway
b. Test to see what “feeds” what
i. Substances secreted are the ones that the organism can possibly produce with
its mutation
IV. Chapter 7 – Enzymes
a. Be able to draw free energy change curves in energy-yielding rxn
b. Definitions:
i. Activation energy: minimum energy required to enter a transition state
ii. Transition state: an unstable state in which the molecules can then
spontaneously undergo an exergonic reaction
iii. Net reaction energy change: the net rxn energy change
c. Enzymes lower Ea
d. E+S <-> ES <-> EP <-> E+P
i. Meaning that the enzyme makes the transition state, and rxn is reversible
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e. Key terms explaining saturation:

i. Saturation, linear, free enzyme molecules, substrate concentration, enzyme
availability limits rate of rxn, enzyme-substrate/enzyme-product complexes
f. Law of mass action
i. States that rate of an uncatalyzed chem. Rxn is directly proportional to
concentrations of reactants.
1. i.e. + [reactants] = + rate uncatalyzed reaction
2. linear
g. Buffers
i. Substance resistant to pH change by adding or removing H+ depending on the
pH of solution
h. Enzymes denature with pH far from optimum
i. Analyzing concentration-velocity graphs:
i. Vm = maximal velocity
ii. Km = Michaelis-Menten constant
1. The substrate concentration required to reach one-half of max velocity
2. Representation of enzyme efficiency
3. Some enzymes have lower Km for same velocity as another enzyme
iii. More enzyme concentration makes same Km but higher Vm
1. Same enzyme thus same efficiency
iv. Competitive inhibitor makes shift in Km to the right a bit because more substrate
required to make shift in equilibrium
1. i.e. competitive inhibitor is only temporary, so more substrate = more
possibility of collision
v. Non-competitive inhibitor only makes shift in Vm to become lower
1. Non-competitive inhibitor reduces molecules of functioning enzymes
V. Chapter 8 – Osmosis and Membrane Permeability
a. Diffusion: the movement of a substance from an area of high concentration to an area
of lower concentration, i.e. area of more negative psi to area of more positive psi across
a selectively permeable membrane
b. Osmosis: the movement of water from area of more positive psi to area of more
negative psi across selectively permeable membrane
c. Water potential
i. 0 at atmospheric conditions in pure water
ii. Ψ = Ψp + Ψs
1. Related to physical pressure and solute pressure
2. Represented in MPa = 10 bar ≈ 10 atm
3. Inside a plant cell, physical pressure is turgidity, solute pressure is solute
d. Osmolarity
i. Measures number of total solute particles per liter
ii. for example, we would expect 0.10M NaCl to be 0.20M NaCl
1. however due to interactions between molecules, they become less
a. close to 0.187M ish
e. Partition coefficient
i. The quotient of solubility in lipid divided by solubility in water
ii. Solute’s rate of diffusion through membrane directly related to lipid/water
coefficient
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iii. Thus with a very small P.C., substance will not be able to diffuse across
membrane readily
1. i.e. ions will have small P.C.
iv. Take methanol, ethanol, propanol
1. Increasing carbon chain lengths
2. Relatively same solubility due to –OH, but lipid solubility also increases
towards propanol
3. Therefore propanol has fastest solubility across membrane
f. Hemolysis = rupture of erythrocyte membrane
g. What was done in the experiment:
i. Isotonic solution of different solute than what’s present in erythrocyte
suspension was placed together
1. Because different solutes, the solute moved across cell membrane to
change negatively Ψs in the cell, thus triggering osmosis into the cell
a. Thus hemolysis
h. Statistics stuff
i. Population is the overall stuff we’re getting things from, i.e. the erythrocytes
provided in lab
ii. Sample: random stuff we’re picking from population, ii.e. erythrocytes used
iii. Replicate: repeated samples in the test
iv. Parameter: quantifiable characteristic of population of things
v. Statistic: single value obtained by parameter replicates, or computed