Presentations/Assays

Question 1

X-Ray Diffraction, how does it collect info? (S)

Answer 1

transitions in inner and outer electrons

Question 2

X-Ray diffraction, what info can it collect? (S)

Answer 2

Identifies atomic and molecular structure including crystalline structure

Question 3

X-Ray Diffraction is used with? (S)

Answer 3

Powdered metals/ceramics

Question 4

UV-Vis, how does it collect info? (S)

Answer 4

transitions in valence electrons

Question 5

UV-Vis, what info can it collect? (S)

Answer 5

Identifies chemical groups in sample, can also be used to quantify amount of a compound in a solution that adsorbs at those wavelengths

Question 6

UV-Vis is used with? (S)

Answer 6

Polymers mainly

Question 7

IR, how does it collect info? (S)

Answer 7

Vibrations in bonds, similar to UV-Vis but different wavelengths

Question 8

IR, what info can it collect? (S)

Answer 8

Identifies chemical groups in sample, can also be used to quantify amount of a compound in a solution that adsorbs at those wavelengths

Question 9

Raman, how does it collect info? (S)

Answer 9

vibrations in bonds (Stokes/anti-stokes) through energy scattered and changes in polarizability

Question 10

Raman, what info can it collect? (S)

Answer 10

Identify compounds and study their structure, not used with metals. Can be coupled with microscopy

Question 11

NMR, how does it collect info? (S)

Answer 11

Changes in nuclear spin with different radio frequencies

Question 12

NMR, what info can it collect? (S)

Answer 12

Chemical composition of polymers and biomolecules

Question 13

Mass Spectrometry measures?

Answer 13

ion mass-to-charge-ratio

Question 14

SEM uses _____ for _____?

Answer 14

Scattered electrons, imaging/morphology. Stands for scanning electron microscopy

Question 15

Mechanical property tests? (4)

Answer 15

Tensile, compression, shear, and bending

Question 16

DSC, what two modes? What does it measure?

Answer 16

Heat flux (same power measure differences in temp) and power compensation (same temp measure changes in energy required to keep at that temp). Measures Tm, Tg, heat capacity etc

Question 17

HPLC stands for? Two types? Can measure what?

Answer 17

Hig performance liquid chromatography, Affinity chromatography and size exclusion, molecular weight

Question 18

ATR-FTIR stands for? is a form of? can measure?

Answer 18

Attenuated total reflection, Fourier transform IR, IR, measures energy absorbed, vibrations in bonds and change in dipole moment (Optics with internal relfection)

Question 19

XPS/ESCA how does it collect info?

Answer 19

inner electrons emitted due to energy absorbed

Question 20

SEM produces ___ image while TEM produces ___ image

Answer 20

2D/3D, 2D

Question 21

TEM uses ___ for ___?

Answer 21

transmitted electrons, morphology of nanoparticles and thin sections. Stands for transmission electron microscopy

Question 22

SPM/AFM stands for ? Measures? Used to get?

Answer 22

Scanning Probe Microscopy/ Atomic Force Microscopy, , atomic forces between tip and surface to get an image, contact mode/dynamic mode

Question 23

Protein adsorption measured in 3 ways? list them

Answer 23

AT-FTIR (Change in optical properties based on layer thickness), QCM (Change in mass which affect frequency of crystal), Colorimetric/Florescence Assays (Dying/label the protein)

Question 24

Identity of protein measured in 3 ways? list them.

Answer 24

Mass Spectrometry, western Blotting, Elisa (need to know what protein searching for for wb and elisa)

Question 25

Viability Assays 4 types? list them, tell what they detect, and explain why each works

Answer 25

Trypan blue (damaged/dead cells, dye that passes through damaged cell emembrane), Propidium Iodide (dead cells, can pass through damaged membrane, binds to nuclei, emits red light) Calcein-AM (live cells, dye enzymatically converted to fluorescent calcein), MTT (Live cells, measures metabolic activity, converted to formazan which is dark blue)

Question 26

Methods of counting cells? (3)

Answer 26

hemocytometer, automated counting systems, microscopes

Question 27

Proliferation assay?

Answer 27

BrDU, analog of thymidine, cells proliferatie and new DNA has BrDU, detected using fluroescent antibody of BrDU. Has to be coupled with a quantification method (fluorescence microscopy)

Question 28

Cytotoxicity assays? (2)

Answer 28

Contact and elution (combined with viability and counting method)

Question 29

Injury assay?

Answer 29

LDH, lactate dehydrogenase, LDH leaks from damages cells coverts tetrazolium salt to formazan

Question 30

Migration 2 bad ways, 2 good ways.

Answer 30

Bad: ring barrier test, capillary tube test. Good: microscopic tracking, Boyden chamber assay

Question 31

Protein production measured by?

Answer 31

Western blotting, Elisa, Immunocytochemistry (previous 3 require antibody) Mass spectrometry colorimetric/florescence assays

Question 32

PCR/RT-PCR. What's the difference? Which is used for gene expression?

Answer 32

Very similar but RT-PCR first transcribes mRNA to ssDNA then normal steps occur. RT-PCR is used because mRNA represents a gene thats being expressed. If PCR used, it only shows if gene is there, not if it is expressed.

Question 33

What are the steps of Elisa?

Answer 33

- -Primary antibody attached to a surface> - -Sample is added and protein of interest binds to antibody> - -Secondary antibody binds to another region n the protein> - -Secondary antibody, which is also an enzyme, enzymatically converts a molecule and color change is measured

Question 34

What are the steps of western blotting?

Answer 34

- -Sample treated with sodium dodecyl sulfate (SDS)> - -SDS-treated sample loaded into SDS containing poly(acrylamide) gel (SDS-PAGE) and exposed to electrical field> - -SDS is negative so molecules move to positive pole, separated by molecular weight> - -Blotted onto a membrane and washed with milk to block non-specific binding> - -Primary antibody on protein added> - -Labeled secondary antibody added that binds to primary antibody - -Secondary antibody can also be enzyme conjugated and change in color can be monitored