If there are two different Nps prepared for two different drugs encapsulated in a single polymeric system(co-delivery) then can the uptake of both the NPs detected using confocal microscopy? In this case two types of fluorescent dyes needed ??
In another case, for cellular localization expt which dyes except Lysotracker can be used?
Chittaranjan. B.
LysoTracker Red DND-99, selectively accumulated in cellular compartments with low internal pH, can be used to study biosynthesis and pathogenesis
lysosomes and cell detection. LysoTracker Red DND-99 easily penetrates cells, accumulates in lysosomes and can be used at very low concentrations. In addition to this fluorescent dye, MitoTracker RED CMXRos, Hoechst 33342 can be used for cell detection.
Modern confocal microscopy allows us to solve three main tasks: the study of the fine structure of a cell, the spatial relationship of two or more substances (nanoparticles) in the cell, as well as the study of dynamic processes occurring in living cells.
Dear Chittaranjan. B. , the simple and general answer to your first question should be clearly negative, the cellular uptake and cellular location of NPs are different experiments. However, they could be the same experiment if all the uptook NP´s are localized at the same organelle or cell region.
With respect to your second question, if you have two different drugs encapsulated in a polymeric delivery system to facilitate the uptake by the cell, they could be detected by confocal microscopy if these two drugs show distinctive fluorescence signals. It means that these two drugs must show auto-fluorescence in the conditions of excitation used and they must show different "colours" to be distinguished. If one or both drugs do not show fluorescence under the working conditions or they show the same or very close colours, then you must use some dye to label each drug. If your delivery system is intended for reaching some cell´s organelle and just there delivering its cargo, may be it could be possible to just follow the whole system (drug A + drug B + polymeric system) and labelling it with the addition of some fluorescent dye, well inside or attached to the polymeric shell. In this way you could see if the whole system reaches the cell and then the target organelle. To use this methodology you need to be sure that both drugs reach the target and that the polymeric shell keeps its cargo till the destination, in other case you would be following an empty- fluorescent- shell.
Hope this helps. Good luck with your research work.
- Badges
- Science topic