These microscopes operate by measuring the fluorescent light that some compounds emit naturally or the light emitted by artificial fluorophores
, and by exploiting various quantum properties of the fluorophore
, can deliver a resolution smaller than the one imposed by the diffraction limit.
(1,4,5) Immunofluorescence has a higher sensitivity and allows for the colocalization of several antigens labeled by different fluorophores
compared with immunohistochemistry.
The main applications of FLIM are studies of molecular interaction by Forster Resonance Energy Transfer (FRET), biosensing microenvironmental changes and separation of fluorophores
with overlapping spectral proprieties.
This has led to the use of fluorophores
like 5-ALA (5-aminolaevulinic acid) that highlight the tumour and its margins.2 The use of 5-ALA showed a significant increase in the rate of GTR for MGs.2 However, its use remains limited due to lack of availability and a high cost.
As it is currently accepted, the fluorescence of aquatic CDOM consists of two main bands, the so-called protein-like and humic-like fluorescence coming from two main groups of fluorophores
Galezowski, "Strongly emitting fluorophores
based on 1-azaperylene scaffold," Journal of Organic Chemistry, vol.
A wide range of small molecule fluorophores
and conjugated polymers have been developed for effective sensing, in vapor/solid phase or in organic solvents but not in water.
'This is very different from fluorophores
found in other vertebrates, which are usually proteins or polyenic chains,' Maria Gabriella Lagorio, a photochemist and co-author of the paper, told Chemistry World.
This is readily handled "behind the scenes" by mixing the unrelated primer and probe sets in one reaction, as long as the probe fluorophores
are at distinguishably different wavelengths so that the real-time PCR instrument observes them independently.
The scanner has the ability to scan a 1 cm3 specimen for 20 fluorophores
However, an experimental validation of the suitability of these fluorophores
to retain their selectivity when used simultaneously has not been reported.