Flexafm 5 nir scan head

The FluidFM setup is composed of a FlexAFM 5-NIR scan head controlled by a C3000 controller (Nanosurf, Switzerland), a digital pressure controller (ranging from −800 mbar to +1,000 mbar) and Microfluidic Probes (Cytosurge). The scan head is mounted on an inverted AxioObserver microscope equipped with a temperature-controlled incubation chamber (Zeiss). The microscope is coupled to a spinning disk confocal microscope (Visitron, Germany) with a Yokogawa (Japan) CSU-W1 scan head and an EMCCD camera system (Andor, UK). For all images and videos, a 63× oil objective with 1.4 numerical aperture and a 2× lens switcher was used (without lens switcher: 4.85 pixel/micron and 9.69 pixel per micron with lens switcher); images are in 16 bit format. Image acquisition was controlled using the VisiView software (Visitron); linear adjustments and video editing were made with Fiji [67 (link)]; additionally, images and videos were noise-filtered using the wiener noise filtering function (wiener2; 3 by 3 neighborhood size) in MATLAB R2018a (MathWorks, USA). Movies were created using a self-written MATLAB script in order to visualize several sections or channels within the same movie. Colormaps originate from Thyng and colleagues [68 ]. Images of cantilevers containing extracts (Fig 2) were created summing up the slices of a Z-stack via Fiji and reconverting the image to 16 bit format.