Preparation and characterization of poly(lipid)-coated, fluorophore-doped silica nanoparticles for biolabeling and cellular imaging.
The fabrication, characterization, and implementation of poly(lipid)-coated, highly luminescent silica nanoparticles as fluorescent probes for labeling of cultured cells are described. The core of the probe is a sol-gel-derived silica nanoparticle, 65-100 nm in diameter, in which up to several thousand dye molecules are encapsulated (Lian, W.; et al. Anal. Biochem. 2004, 334, 135-144). The core is coated with a membrane composed of bis-sorbylphosphatidylcholine, a synthetic polymerizable lipid that is chemically cross-linked to enhance the environmental and chemical stability of the membrane relative to a fluid lipid membrane. The poly(lipid) coating has two major functions: (i) to reduce nonspecific interactions, based on the inherently biocompatible properties of the phosphorylcholine headgroup, and (ii) to permit functionalization of the particle, by doping the coating with lipids bearing chemically reactive or bioactive headgroups. Both functions are demonstrated: (i) Nonspecific adsorption of dissolved proteins to bare silica nanoparticles and of bare nanoparticles to cultured cells is significantly reduced by application of the poly(lipid) coating. (ii) Functionalization of poly(lipid)-coated nanoparticles with a biotin-conjugated lipid creates a probe that can be used to target both dissolved protein receptors as well as receptors on the membranes of cultured cells. Measurements performed on single nanoparticles bound to planar supported lipid bilayers verify that the emission intensity of these probes is significantly greater than that of single protein molecules labeled with several fluorophores.
Senarath-Yapa
Muditha D
MD
Department of Chemistry, University of Arizona, Tucson, AZ 85721-0041, USA.
Phimphivong
Sam
S
Coym
Jason W
JW
Wirth
Mary J
MJ
Aspinwall
Craig A
CA
Saavedra
S Scott
SS
D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry*; D008163:Luminescent Measurements / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry*; D017646:Organosilicon Compounds / Q000138:chemical synthesis / Q000737:chemistry; D010316:Particle Size; D011108:Polymers / Q000737:chemistry; D012680:Sensitivity and Specificity; D012822:Silicon Dioxide / Q000737:chemistry*; D013194:Staining and Labeling; D013499:Surface Properties
D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D017646:Organosilicon Compounds; D011108:Polymers; D012822:Silicon Dioxide