Figure 5: Oscilloscope measurements of sample illumination time. Experiments were conducted on a Leica DMI6000B inverted microscope equipped with a Quo- rum WaveFx-X1 spinning disk confocal system running MetaMorph software (version 7.10.2.240). A 561 nm laser was electronically triggered with an AOTF crystal. Interval imaging resulted in ~42ms IO and ~100ms delay between images. Stream acquisition resulted in ~17ms delay between images (in the form of IO).
Figure 6: (A) MCF7 cells were seeded onto fibronectin-coated dishes and stained with LysoTracker™ Green DND-26. Images were collected on the spinning disk confocal microscope using stream acquisition and a 63×1.4 NA oil immersion objective lens. A 491 nm laser (~0.02mW) was used to excite LysoTracker™. Camera exposure time was set to 200ms. Scale bars are 10 µm for whole-cell images and 2 µm for magnified images. (B,C) Lysosome positions were tracked and quantified for speed and persistence. Frequency distributions show pooled data for lysosomes from three cells. N indicates the number of lysosomes analyzed.
