The goal of this study was to develop an innovative approach to image gene expression in intact 3D tissues. Imaging gene expression of individual cells in 3D tissues is expected to have a significant impact on both clinical diagnostic applications and fundamental biological science and engineering applications in a laboratory setting. To achieve this goal, we have developed an integrated approach that combines: 1) microneedle-based minimally invasive intra-tissue delivery of oligonucleotide probes and Streptolysin O (SLO) or CPP; 2) SLO as a pore forming permeation enhancer to enable intracellular delivery of oligonucleotide probes and CPP peptides can also transport conjugated cargo in cells; and 3) fluorescence resonance energy transfer (FRET) pair of ON probes to improve specificity and sensitivity of RNA detection in tissue models. The results of this study demonstrate uniform coating and rapid release of ON probes from microneedles in a tissue environment. Microneedle assisted delivery of ON probes in 3D tissue does not result in cell damage and the ON probes are uniformly delivered in the tissue. The results also demonstrate the feasibility of FRET imaging of ON probes in 3D tissue and highlight the potential for imaging 28-s rRNA in individual living cells.

Date Published: 30 May 2013
PDF: 7 pages
Proc. SPIE 8725, Micro- and Nanotechnology Sensors, Systems, and Applications V, 872512 (30 May 2013); doi: 10.1117/12.2018135

Published in SPIE Proceedings Vol. 8725:
Micro- and Nanotechnology Sensors, Systems, and Applications V
Thomas George; M. Saif Islam; Achyut K. Dutta, Editor(s)