Bio 331 Exam Solved
QUESTIONS AND VERIFIED
CORRECT ANSWERS
[LATEST] GRADED A+ 100%
GUARANTEES SUCCESS
z-stack - CORRECT ANSWER-series of optical solutions taken at different depths
z-stack projection - CORRECT ANSWER-reconstructed image by overlaying many individual z-
stack images
z-stack can be applied to: - CORRECT ANSWER-DIC, brightfield, or fluorescence microscopy
z-stack works best with - CORRECT ANSWER-fluorescence
confocal microscopy - light source - CORRECT ANSWER-laser
confocal microscopy - how light focused? - CORRECT ANSWER-2 confocal pinholes
confocal microscopy - mirror type - CORRECT ANSWER-dichroic mirror
,dichroic mirror purpose - CORRECT ANSWER-bends light from laser towards specimen but
allows through light from fluorophores
2nd pinhole purpose - CORRECT ANSWER-only allows focused light to receptor
1st pinhole purpose - CORRECT ANSWER-focuses light from laser
multi-photon imaging - CORRECT ANSWER-use 2+ photons of lower energy
must hit very fast after one another
less damaging to sample bcz using a longer wavelength
GFP - CORRECT ANSWER-green fluorescent protein
GFP discovery - CORRECT ANSWER-1990s
jellyfish Aequorea victoria
structure that allows chromophore formation - CORRECT ANSWER-Beta barrel
(not an enzyme)
how to get GFP to target specific locations - CORRECT ANSWER-add aa seq to GFP at DNA level
how to fuse GFP to protein of interest - CORRECT ANSWER-at DNA level, before translation
GFP used to: - CORRECT ANSWER-report gene expression
tag cell types/tissues
,how to get diff colours from GFP - CORRECT ANSWER-alter aa sequence of GFP
red-orange fluorophore found in: - CORRECT ANSWER-marine anemone and coral reefs
FRET - CORRECT ANSWER-Fluorescence Resonance Energy Transfer
how FRET works - CORRECT ANSWER-tag 2 molecules with 2 colours of fluor proteins
if close enough, flour1 excites fluor2
see 2nd colour? = close proximity
FRET shows _____, not _____ - CORRECT ANSWER-shows proximity, not binding
photoactivation - CORRECT ANSWER-inactive photosensitive fluor precursor in cage
laser/UV breaks cage and activates fluor
use of photoactivation - CORRECT ANSWER-view movement of fluor through cell after cage
broken
FRAP - CORRECT ANSWER-Fluorescent Recovery After Photobleaching
how FRAP works - CORRECT ANSWER-label all of certain proteins with fluor
photobleach certain region
recovery of fluor in region = movement of proteins into region
what is added in FRAP to prevent new protein synth - CORRECT ANSWER-cycloheximide
, reasons for no fluorescence recovery in FRAP? - CORRECT ANSWER-tight junctions prevent
diffusion btw apical, basal, or lateral sides of cell
protein-protein aggregates
intra/extra cellular tethers (ie cytoskeleton)
interactions with other cells
genetically encoded fluorescent biosensors: purpose - CORRECT ANSWER-determine protein
activity by fusing fluor to protein of interest
genetically encoded fluorescent biosensors: mechanism/detection - CORRECT ANSWER-addition
of signal/activation -> conf change -> brings 2 dif coloured FP close together
= colour change due to FRET
Ca indicators - CORRECT ANSWER-FP whose spectra is altered by Ca binding
intracellular pH indicators - CORRECT ANSWER-FP with emission spectra altered by pH
in microscopy, objective lens is always - CORRECT ANSWER-closest to specimen
electron microscopy types - CORRECT ANSWER-TEM
SEM
TEM stands for - CORRECT ANSWER-transmission electron microscopy
TEM - CORRECT ANSWER-uses e- from above (instead of photons)
specimen on metal grid
QUESTIONS AND VERIFIED
CORRECT ANSWERS
[LATEST] GRADED A+ 100%
GUARANTEES SUCCESS
z-stack - CORRECT ANSWER-series of optical solutions taken at different depths
z-stack projection - CORRECT ANSWER-reconstructed image by overlaying many individual z-
stack images
z-stack can be applied to: - CORRECT ANSWER-DIC, brightfield, or fluorescence microscopy
z-stack works best with - CORRECT ANSWER-fluorescence
confocal microscopy - light source - CORRECT ANSWER-laser
confocal microscopy - how light focused? - CORRECT ANSWER-2 confocal pinholes
confocal microscopy - mirror type - CORRECT ANSWER-dichroic mirror
,dichroic mirror purpose - CORRECT ANSWER-bends light from laser towards specimen but
allows through light from fluorophores
2nd pinhole purpose - CORRECT ANSWER-only allows focused light to receptor
1st pinhole purpose - CORRECT ANSWER-focuses light from laser
multi-photon imaging - CORRECT ANSWER-use 2+ photons of lower energy
must hit very fast after one another
less damaging to sample bcz using a longer wavelength
GFP - CORRECT ANSWER-green fluorescent protein
GFP discovery - CORRECT ANSWER-1990s
jellyfish Aequorea victoria
structure that allows chromophore formation - CORRECT ANSWER-Beta barrel
(not an enzyme)
how to get GFP to target specific locations - CORRECT ANSWER-add aa seq to GFP at DNA level
how to fuse GFP to protein of interest - CORRECT ANSWER-at DNA level, before translation
GFP used to: - CORRECT ANSWER-report gene expression
tag cell types/tissues
,how to get diff colours from GFP - CORRECT ANSWER-alter aa sequence of GFP
red-orange fluorophore found in: - CORRECT ANSWER-marine anemone and coral reefs
FRET - CORRECT ANSWER-Fluorescence Resonance Energy Transfer
how FRET works - CORRECT ANSWER-tag 2 molecules with 2 colours of fluor proteins
if close enough, flour1 excites fluor2
see 2nd colour? = close proximity
FRET shows _____, not _____ - CORRECT ANSWER-shows proximity, not binding
photoactivation - CORRECT ANSWER-inactive photosensitive fluor precursor in cage
laser/UV breaks cage and activates fluor
use of photoactivation - CORRECT ANSWER-view movement of fluor through cell after cage
broken
FRAP - CORRECT ANSWER-Fluorescent Recovery After Photobleaching
how FRAP works - CORRECT ANSWER-label all of certain proteins with fluor
photobleach certain region
recovery of fluor in region = movement of proteins into region
what is added in FRAP to prevent new protein synth - CORRECT ANSWER-cycloheximide
, reasons for no fluorescence recovery in FRAP? - CORRECT ANSWER-tight junctions prevent
diffusion btw apical, basal, or lateral sides of cell
protein-protein aggregates
intra/extra cellular tethers (ie cytoskeleton)
interactions with other cells
genetically encoded fluorescent biosensors: purpose - CORRECT ANSWER-determine protein
activity by fusing fluor to protein of interest
genetically encoded fluorescent biosensors: mechanism/detection - CORRECT ANSWER-addition
of signal/activation -> conf change -> brings 2 dif coloured FP close together
= colour change due to FRET
Ca indicators - CORRECT ANSWER-FP whose spectra is altered by Ca binding
intracellular pH indicators - CORRECT ANSWER-FP with emission spectra altered by pH
in microscopy, objective lens is always - CORRECT ANSWER-closest to specimen
electron microscopy types - CORRECT ANSWER-TEM
SEM
TEM stands for - CORRECT ANSWER-transmission electron microscopy
TEM - CORRECT ANSWER-uses e- from above (instead of photons)
specimen on metal grid
