Oct 4 was not detected with MB in these cells and signal was not increased over time suggesting that MB are generally stable inside the cells. MAP2 and tyrosine hydroxylase mRNAs were expressed 2 and 3 days post induction of differentiation, respectively. Nestin and NeuN mRNA were expressed in approximately 35% of the LUHMES cells grown in growth medium, and in 80–90% of cells after differentiation. Gene expression was detected at different time points using fluorescence microscopy. The cells will then each contain about 60,000 MBs.

Mathematical modeling, simulations and experiments indicated that MB concentration was equal to the MB in the transfection medium after 10 min transfection. MBs were transfected into human mesencephalic cells (LUHMES) using streptolysin-O-based membrane permeabilization. In order to monitor gene expression during nerve cell development, molecular beacon (MB) probes targeting markers representing different stages of neuronal differentiation were designed and synthesized as 2'-O-methyl RNA backbone oligonucleotides. Monitoring gene expression is an important tool for elucidating mechanisms of cellular function. 2Center for Individual Nanoparticle Functionality, Technical University of Denmark, Kgs.1Department of Micro- and Nanotechnology, Technical University of Denmark, Kgs.Mirolyuba Ilieva 1, Paolo Della Vedova 1, Ole Hansen 1,2 and Martin Dufva 1 *

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