RNA Detection

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3.3 Cellular Delivery
of MB–Dextran
Mixtures


dextran into a large number of cells with high viability, as shown in
our previous studies [18, 19].
Day 1:


  1. Coat the glass-bottom wells of the 8-well chambered cover
    glass by adding 250 μL fibronectin per well and incubate
    overnight at 37C(seeNote 10).

  2. Seed cells in T-25 flasks in DMEM growth media without
    phenol red or antibiotics so that they will be no more than
    70% confluent on the day of the experiment (seeNote 11).


Day 2:


  1. Aspirate the media from the cells and rinse the flask using 5 mL
    of prewarmed 1PBS for about 2 min.

  2. Aspirate the PBS and add 1 mL of phenol red-free trypsi-
    n–EDTA; incubate for 1 min at room temperature.

  3. Aspirate half of the trypsin (about 500μL) and incubate at 37C
    to detach all of the cells from the flask surface (seeNote 12).


Fig. 1Fluorescent microscopy images of microemulsion bubbles containing MBs and IRDye®800-labeled
dextran in the presence and absence of nucleic acid targets.FMBdenotes the fluorescence signals of the MB.
FREFdenotes the fluorescence signal of the dextran (scale bar, 10μm)


248 Mingming Chen et al.

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