| Method |
FRAP: To measure export kinetics in nuclear envelopes more precisely, researchers employed photobleaching (Keminer and Peters, 1999). After the addition of transport media containing either 15 µM GFP or 7.4 µM Alexa488 NTF2 dimers to a filter-attached nuclear envelope, the transport substrate was allowed to equilibrate between the OSTR chamber and filter pores. Then, employing the time-lapse/bleaching program of the confocal microscope system together with a high-power objective (100-fold, 1.3 numerical aperture), a prebleach scan was acquired, a filter pore selected for 1 s of photobleaching, and the recovery of the fluorescence of the photobleached filter pore followed by a series of scans. |