MS: ✗
BD: ✗
Fluorescence Optics (488 nm)
Generally enhances the dynamic rane by extending into the low end of sample concentration into the low nano molar, depending on labeling efficiency and fluorophore quantum yield.
the minimal FDS power setting still enable the use of low-micromolar concentrations
Zhao et al 2014 report that fluorescent optics can provide uable data for concentrations as low micro-molar.
LABELING REQUIREMENTS: fluorphores with 488-nm exitation maxima are ideal to use, but other dyes can be used if they have higher final concentrations for producing a singal equivalent to samples with the 488-nm excitation maxima.
When labing native functional groups, make sure that its behavior is equivalent to that of the unlabelled protein.
When to Use
Pros and Cons
Advantages Exquisite selectivity to measure tagged molecule and can be used against an impure background
Very high sensitivity (down to pM concentrations)
No refractive artifacts
Fast scanning speed (all cells and channels are scanned simultaneously)
No buffer interference
Great for hetero-interactions, in-vivo studies, GFP fusions proteins.
Disadvantages
Relatively high stochastic noise
Most samples must be tagged by a fluoropore.