title: Optical delivery of liposome encapsulated chemical stimuli to neuronal cells.
abstract: Spatially confined and precise time delivery of neuroactive molecules is an important issue in neurophysiology. In this work we developed a technique for delivering chemical stimuli to cultured neurons consisting in encapsulating the molecules of interest in liposomes. These vectors were then loaded in reservoirs consisting of glass capillaries. The reservoirs were placed in the recording chamber and single liposomes were trapped and transported out by optical tweezers to the site of stimulation on cultured neurons. Finally, the release of liposome content was induced by application of UV-pulses, breaking the liposome membrane. The efficiency of encapsulation and release were first evaluated by loading the liposomes with fluorescein. In order to test the effect of the UV-induced release, liposomes with diameter ranging from 1 to 10 μm (fL to pL volumes), were filled with KCl and tested on neuronal cells. Neuronal cultures, loaded with Ca(2+) dye, were monitored by imaging intracellular Ca(2+). An efficient release from the liposomes was demonstrated by detectable calcium signals, indicating stimulated depolarization of the neuronal cells by KCl. The present technique represents an alternative method for focal chemical stimulation of cultured cells that circumvents some of the limitations of microejection and photorelease of caged compounds.
authors.affiliation: CNR, Istituto Officina dei Materiali, Laboratorio TASC, Ed. MM, Area Science Park- Basovizza, SS 14, Km 163.5, 34149, Trieste, Italy. pinato@tasc.infm.it
authors.forename: Giulietta
authors.initials: G
authors.lastname: Pinato
authors.forename: Tiziano
authors.initials: T
authors.lastname: Raffaelli
authors.forename: Elisa
authors.initials: E
authors.lastname: D'Este
authors.forename: Federica
authors.initials: F
authors.lastname: Tavano
authors.forename: Dan
authors.initials: D
authors.lastname: Cojoc
mesh_terms: D000818:Animals; D002118:Calcium; D002478:Cells, Cultured; D016503:Drug Delivery Systems / Q000295:instrumentation / Q000379:methods*; D019793:Fluorescein; D006624:Hippocampus / Q000166:cytology; D007834:Lasers; D008081:Liposomes / Q000737:chemistry*; D009474:Neurons / Q000737:chemistry* / Q000166:cytology; D052898:Optical Tweezers; D010316:Particle Size; D010782:Photolysis; D011189:Potassium Chloride; D051381:Rats
chemical_list: D008081:Liposomes; D011189:Potassium Chloride; D002118:Calcium; D019793:Fluorescein