For telomere imaging in fixed cells, the 4f system consisted of two f=15 cm lenses (Thorlabs), a linear polarizer (Thorlabs) to filter out the light that is polarized in the unmodulated direction of the LC-SLM, a 1920 × 1080 pixel LC-SLM (PLUTO-VIS, Holoeye) and a mirror for beam-steering. A sCMOS camera (Prime95B, Photometrics) was used to record the data. The sample was illuminated with 561 nm fiber-coupled laser-light source (iChrome MLE, Toptica). The excitation light was reflected up through the microscope objective by a multibandpass dichroic filter (TRF89902-EM - ET - 405/488/561/647 nm Laser Quad Band Set, Chroma). Emission light was filtered by the same dichroic and also filtered by another 617 nm band pass filter (FF02-617/73 Semrock).
For volumetric telomere tracking in live cells, images were recorded with an EMCCD camera (Andor iXON), exposure time 100 ms and EM-gain 170. The sample was illuminated at ≈2 kW/cm2 with 561 nm light from a fiber-coupled laser (iChrome MLE, Toptica). All movies were recorded for 50 s (500 frames).
For volumetric telomere tracking in live cells, images were recorded with an EMCCD camera (Andor iXON), exposure time 100 ms and EM-gain 170. The sample was illuminated at ≈2 kW/cm2 with 561 nm light from a fiber-coupled laser (iChrome MLE, Toptica). All movies were recorded for 50 s (500 frames).