A continuous-wave laser centered at 445 nm (iBeam smart, Toptica) is collimated and focused onto the back focal plane of an oil-immersion microscope objective (α Plan-Apochromat ×100, NA 1.46, Zeiss). A coverglass is positioned at the focus of the microscope objective using a piezo positioner (Nano-LPQ, Mad City Labs). The iSCAT field is imaged using a scientific CMOS camera (MV1-D1024E-160-CL, Photonfocus).
TIRF illumination was done with a laser beam at 631 nm, which was directed into the iSCAT pathway via a dichroic mirror (D1, Chroma ZT647rdc-UF3) mounted on a translation stage and a second dichroic mirror (D2, Chroma T480spxxr-UF3). The fluorescence signal was collected via the same microscope objective that was used for the iSCAT measurements. D2 separated the fluorescence from the iSCAT path and transmitted it through D1 onto a CCD (charge-coupled device) camera (Hamamatsu Orca Flash). Here, we also used a band pass filter (ET700/75) in front of the camera (S1).
TIRF illumination was done with a laser beam at 631 nm, which was directed into the iSCAT pathway via a dichroic mirror (D1, Chroma ZT647rdc-UF3) mounted on a translation stage and a second dichroic mirror (D2, Chroma T480spxxr-UF3). The fluorescence signal was collected via the same microscope objective that was used for the iSCAT measurements. D2 separated the fluorescence from the iSCAT path and transmitted it through D1 onto a CCD (charge-coupled device) camera (Hamamatsu Orca Flash). Here, we also used a band pass filter (ET700/75) in front of the camera (S1).
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