RNA Detection

adjacent frames. The resolution of the images we typically

acquire are 120 nm120 nm800 nm at 3.3–5 Hz (see

Note 16).

4 Notes

1. The dye should be at least 2–3 nucleotides away from the
   terminus.


2. The temperature of measurement should be the same as the
   temperature of the in vivo experiments.


3. To obtain ovaries that contain all developmental stages and that
   are not in a relative nutrient deficient state, place 5–6 females
   and the same number of males into a fresh vial containing the
   standard food and granular baker’s yeast 16–24 h before
   dissection.


4. The hydrophobic wax covering the cuticle is dissolved by the
   ethanol wash preventing the fly from floating on the surface of
   the dissection buffer. This brief wash does not affect the viabil-
   ity of the ovaries.

Fig. 2InjectedDrosophilaoocyte. (a–a") Z-stack of a Stage 9 oocyte expressing theoskarMS2-MCP-EGFP
reporter system 8 injected with a mixture of three TO-labeled LNA mixmer FIT probes that target
oskarmRNA (magenta,a"). The site of the injection is marked by a microdamage in the follicular epithelium
(arrow,a^0 ). Note that there is a mild aspecific accumulation of the injected probes in the nuclei (e.g., nurse
cells o the left). (b–b") Time projection of 4 s (12 frames) of an in vivo time lapse acquisition.OskarmRNPs are
highlighted with MCP-EGFP (green,a") and TO (magenta,b^0 ).Yellowarrowheads (b) indicate streaks drawn by
mRNPs moving along a linear trajectory. These trajectories are marked by both MCP-EGFP and TO

284 Jasmine Chamiolo et al.