title: Assessing delivery and quantifying efficacy of small interfering ribonucleic acid therapeutics in the skin using a dual-axis confocal microscope.
abstract: Transgenic reporter mice and advances in imaging instrumentation are enabling real-time visualization of cellular mechanisms in living subjects and accelerating the development of novel therapies. Innovative confocal microscope designs are improving their utility for microscopic imaging of fluorescent reporters in living animals. We develop dual-axis confocal (DAC) microscopes for such in vivo studies and create mouse models where fluorescent proteins are expressed in the skin for the purpose of advancing skin therapeutics and transdermal delivery tools. Three-dimensional image volumes, through the different skin compartments of the epidermis and dermis, can be acquired in several seconds with the DAC microscope in living mice, and are comparable to histologic analyses of reporter protein expression patterns in skin sections. Intravital imaging with the DAC microscope further enables visualization of green fluorescent protein (GFP) reporter gene expression in the skin over time, and quantification of transdermal delivery of small interfering RNA (siRNA) and therapeutic efficacy. Visualization of transdermal delivery of nucleic acids will play an important role in the development of innovative strategies for treating skin pathologies.
authors.affiliation: Stanford University, Department of Electrical Engineering, Ginzton Laboratory, Stanford, California 94305, USA. hra@stanford.edu
authors.forename: Hyejun
authors.initials: H
authors.lastname: Ra
authors.forename: Emilio
authors.initials: E
authors.lastname: Gonzalez-Gonzalez
authors.forename: Bryan R
authors.initials: BR
authors.lastname: Smith
authors.forename: Sanjiv S
authors.initials: SS
authors.lastname: Gambhir
authors.forename: Gordon S
authors.initials: GS
authors.lastname: Kino
authors.forename: Olav
authors.initials: O
authors.lastname: Solgaard
authors.forename: Roger L
authors.initials: RL
authors.lastname: Kaspar
authors.forename: Christopher H
authors.initials: CH
authors.lastname: Contag
mesh_terms: D000818:Animals; D016503:Drug Delivery Systems; D005528:Foot; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal / Q000379:methods*; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D012867:Skin
chemical_list: D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins