Optical nanosensors for chemical analysis inside single living cells. 1. Fabrication, characterization, and methods for intracellular delivery of PEBBLE sensors.
Spherical optical nanosensors, or PEBBLEs (probes encapsulated by biologically localized embedding), have been produced in sizes including 20 and 200 nm in diameter. These sensors are fabricated in a microemulsion and consist of fluorescent indicators entrapped in a polyacrylamide matrix. A generalized polymerization method has been developed that permits production of sensors containing any hydrophilic dye or combination of dyes in the matrix. The PEBBLE matrix protects the fluorescent dye from interference by proteins, allowing reliable in vivo calibrations of dyes. Sensor response times are less than 1 ms. Cell viability assays indicate that the PEBBLEs are biocompatible, with negligible biological effects compared to control conditions. Several sensor delivery methods have been studied, including liposomal delivery, gene gun bombardment, and picoinjection into single living cells.
Clark
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HA
Department of Chemistry, University of Michigan, Ann Arbor 48109-1055, USA.
Hoyer
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M
Philbert
M A
MA
Kopelman
R
R
D002470:Cell Survival; D004396:Coloring Agents; D006801:Humans; D008081:Liposomes / Q000494:pharmacology; D008422:Materials Testing; D008845:Microinjections / Q000295:instrumentation / Q000379:methods; D008854:Microscopy, Electron; D008967:Molecular Biology / Q000379:methods*; D009447:Neuroblastoma / Q000188:drug therapy / Q000473:pathology; D055095:Optics and Photonics / Q000295:instrumentation*; D010316:Particle Size; D019736:Prostheses and Implants; D015203:Reproducibility of Results; D012235:Rhodamines / Q000737:chemistry; D013997:Time Factors; D014407:Tumor Cells, Cultured
D004396:Coloring Agents; D008081:Liposomes; D012235:Rhodamines; C050829:sulforhodamine 101