Figure 3

Electroporation of MBs allows the detection of single molecules and single molecule tracing. Cells were illuminated via HILO to reduce the background fluorescence. A) The overview image shows an electroporated cell. Notice the high fluorescent signal arising from cell debris attached to the cellular membrane. The right panel contains details of a detected single molecule signal in consecutive time frames t0 to t3 (SNR 6,7 ± 2) (5 ms illumination and 30 ms delay between each time frame). B) Heat shock transfection of MBs allows the detection of single molecules. The overview image shows a heat shock treated cell. Since the signal-to-noise-ratio of the detected single molecules is 10.1 ± 0.23, the heat shock performed better than electroporation. The right panel contains details of a detected single molecule signal in consecutive time frames t0 to t3 (5 ms illumination and 30 ms delay between each time frame). The arrowheads points to signals of single molecules and the arrow demarks the signal arising from extracellular debris.