BioAFM methods reporting

Microscope (Make, Model)	Nikon, AXR NSPARC with dual camera dual eTIRF Confocal
Inverted or upright	Inverted
AFM Scan Heads	Select the one/s used: Bruker, CellHesion 300 Bruker, JPK NanoWizard V
Objective: manufacturer, magnification, numerical aperture, immersion, correction (if any)	Select the one/s used: Nikon, 100x Apochromat TIRF, 1.49, oil, collar 0.13-0.2, WD 0.12 Nikon, 100x, 1.45, oil, CFl Plan Apochromat λD, WD 0.13 Nikon, 60x, 1.42, oil, CFl Plan Apochromat λD, WD 0.15 Nikon, 40x, 1.3, oil, CFl Plan Fluor, WD 0.24 Nikon, 20x, 0.8, oil, CFl Plan Apochromat λD, WD 0.8 Nikon, 10x, 0.45, oil, CFl Plan Apochromat λD, WD 4.0
Light source(s)	Select the one/s used: Lasers: AXR NSPARC: LUA-S4 Laser Unit AS405/488/561/640 15mW: 405, 488, 561, 647 nm Dual eTIRF: LUNF 561/640nm 60/50mW FO1,488 nm 60mW FO2: 488, 561, 647 nm.
Detector AX	Tuneable DUX-VB4 detector unit: 3 x GaAsPs + 1x multi alkali PMT Nikon Spatial ARray Confocal (NSPARC)
Scanner AX	Galvano/Resonant Functional 2K: 2k x 2k resonant (operating frequency 7.8KHz) and 8k x 8k galvano scan head
Cameras eTIRF	2x Photometrics 95B Prime sCMOS
Filters, dichroic mirrors, beam splitters	Select the one/s used: AX: DUX-VB4 Dichroic: Quad 405/488/561/640 BS20/80 Emission filters: A1-F 452/45 AX-F 700/75 NSPARC: Emission filters/ Barrier Filters: Quad BA 446/523/600/677 A1-F 452/45 AX-F 525/50 AX-ISM-BA 540/30 AX-F 593/46 AX-F 700/75 AX-ISM-BA 607/LP TIRF: Quad: Chroma 405/488/561/640 Emission filters: DAPI: Semrock FF02- 447/60 nm BrightLine single-band bandpass eGFP: Semrock FF01-525/30 nm BrightLine single-band bandpass YFP: Semrock FF01-542/27 nm BrightLine single-band bandpass TagRFP: Chroma ET610/20 nm Cy5, SPY700:Semrock FF01-708/75 nm BrightLine single-band bandpass
fpbase.org channel set-up
Additional hardware used, e.g., stage motor, incubation chamber and set-up, Piezo stage or objective	Microscope body, Nikon ECLIPSE Ti2-E Objective lens piezo (200 μm). Mad City Labs Nano-F200S JPK Motorized Precision stage. Travel range > 20 mm x 20 mm JPK acoustic enclosure 37.0°C Okolab Black incubation chamber H201 Bruker CO₂ controller 0-20%
Sample Holders	Select the one/s used: JPK Standard for microscopic slide JPK BioCell for round coverslips (22-25.4 mm) with temperature control JPK PetriDishHeater (35 mm dishes) with humidity sensor and temperature control JPK Basic Fluid Cell (Ring on microscopic slide) JPK Sample holder for small samples (AC-00-02)
Acquisition software and settings	NIS Elements v6.xx.xx JPK SPM
AFM modes	CellHesion 300: Contact Mode Force Spectroscopy, Contact Mode Force Mapping, MicroRheology Force Spectroscopy, MicroRheology Force Mapping, Contact Mode Smart Mapping Nanowizard V: PeakForce Tapping™(PFT imaging-biomolecules in liquid, -cells in liquid), Quantitative Imaging (QI imaging, advanced imaging, -biomolecules in liquid, -cells in liquid), PeakForce-QI, Contact mode imaging, AC mode imaging, Detail the scan size, scan rate, force setpoint, and oscillation amplitude.
AFM probe	Manufacturer and model of the cantilever, its resonance frequency, spring constant, and tip radius as well as tip shape. Method used to determine the cantilever's spring constant and the calibration standards.