- Press the injection button on the joystick to inject the target
RNA solution into the cell. The needle will return to the search
level after injection. - Move the microinjection capillary out of the field of view using
the joystick. - Acquire fluorescent images of the cell at different time points
using the Cy5 filter set. Example images are shown in Fig.3 (see
Note 24).
4 Notes
- To quantify the extent of MB nonspecific opening in living
cells, it is important to recognize that MBs can still emit
Fig. 3Functionality analysis of 2Me/PSLOOPand conventional MBs in living cells. Excess complementary RNA
targets were injected into HeLa cells 8 and 24 h after microporation of 2Me/PSLOOP, 2Me/PSFULLor 2Me MBs.
Fluorescent images were acquired before and within several minutes after injection. (a) Representative pre-
and post-injection fluorescent images. (b) 2Me/PSLOOPMBs exhibited greatest signal enhancement upon
injection of the targets into the cells. Each data point represents the meanSD from at least 10 cells.
Asteriskrepresents significant differences (p<0.05) from both 2Me/PSFULLand 2Me MBs (scale bar, 10μm)
(Reproduced from [19] with permission from Elsevier)
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