title: Advanced fluorescence microscopy methods illuminate the transfection pathway of nucleic acid nanoparticles.
abstract: A great deal of attention in biopharmacy and pharmaceutical technology is going to the development of nanoscopic particles to efficiently deliver nucleic acids to target cells. Despite the great potential of nucleic acids for treatment of various diseases, progress in the field is fairly slow. One of the causes is that development of suitable nanoscopic delivery vehicles is hampered by insufficient knowledge of their physicochemical and biophysical properties during the various phases of the transfection process. To address this issue, in the past decade we have developed and applied advanced fluorescence microscopy techniques that can provide a better insight in the transport and stability of nanoparticles in various biological media. This mini-review discusses the basic principles of fluorescence recovery after photobleaching (FRAP), fluorescence correlation spectroscopy (FCS) and single particle tracking (SPT), and gives an overview of studies in which we have employed these techniques to characterize the transport and stability of nucleic acid containing nanoparticles in extracellular media and in living cells.
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Harelbekestraat 72, 9000 Gent, Belgium.
authors.forename: Kevin
authors.initials: K
authors.lastname: Braeckmans
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Harelbekestraat 72, 9000 Gent, Belgium.
authors.forename: Kevin
authors.initials: K
authors.lastname: Buyens
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Harelbekestraat 72, 9000 Gent, Belgium.
authors.forename: Broes
authors.initials: B
authors.lastname: Naeye
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Harelbekestraat 72, 9000 Gent, Belgium.
authors.forename: Dries
authors.initials: D
authors.lastname: Vercauteren
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Harelbekestraat 72, 9000 Gent, Belgium.
authors.forename: Hendrik
authors.initials: H
authors.lastname: Deschout
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Harelbekestraat 72, 9000 Gent, Belgium.
authors.forename: Koen
authors.initials: K
authors.lastname: Raemdonck
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Harelbekestraat 72, 9000 Gent, Belgium.
authors.forename: Katrien
authors.initials: K
authors.lastname: Remaut
authors.affiliation: Laboratory of Gene Therapy, Ghent University, Heidestraat 19, 9820 Merelbeke, Belgium.
authors.forename: Niek N
authors.initials: NN
authors.lastname: Sanders
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Harelbekestraat 72, 9000 Gent, Belgium.
authors.forename: Jo
authors.initials: J
authors.lastname: Demeester
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Harelbekestraat 72, 9000 Gent, Belgium. Electronic address: Stefaan.DeSmedt@UGent.be.
authors.forename: Stefaan C
authors.initials: SC
authors.lastname: De Smedt
mesh_terms: D031541:Fluorescence Resonance Energy Transfer; D008856:Microscopy, Fluorescence / Q000379:methods*; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D009696:Nucleic Acids / Q000008:administration & dosage* / Q000737:chemistry; D038761:Photobleaching; D013050:Spectrometry, Fluorescence; D014162:Transfection / Q000379:methods*
chemical_list: D009696:Nucleic Acids