REAGENT or RESOURCE	SOURCE	IDENTIFIER
Chemicals, peptides, and recombinant proteins
DMEM	Thermo Fisher Scientific	12491015
DMEM without phenol red	Invitrogen	N/A
Antibiotic/antimycotic for IMCD cells	Thermo Fisher Scientific	15240062
DMEM HeLa Cells	Institute of Molecular Biotechnology of the Austrian Academy of Sciences	N/A
penicillin–streptomycin	Sigma-Aldrich	P0781
GlutaMAX	Thermo Fisher Scientific	35050061
rat tail-derived Collagen Type I	Corning	354236
truncated CMV promoter	Addgene	110718
mNeonGreen-Paxillin	Allele Biotechnology	N/A
Alexa Fluor 568	Thermo Fisher Scientific	A20003
Deposited data
Raw test dataset	This paper	https://zenodo.org/record/6881276
Experimental models: Cell lines
Inner medulla collecting duct (IMCD) mouse epithelial cells	ATCC	ATCC: CRL-2123, RRID: CVCL_0429
U2OS female cells	ATCC	ATCC: HTB-96 RRID:CVCL_0042
Sum159O breast cancer cells	Whitehurst68,69	N/A
HeLa Kyoto line	S. Narumiya	RRID:CVCL_1922
Stable Embryonic Stem Cells	Liu56	N/A
Software and algorithms
u-track3D	This paper	https://doi.org/10.5281/zenodo.10055024
MATLAB 2015a-2023a	Mathworks	RRID:SCR_001622
Gmic	D. Tschumperlé/GREYC/CNRS	https://gmic.eu/
AMIRA v6	Thermo Fisher Scientific	RRID:SCR_007353
LabView	Emerson	RRID:SCR_014325
Other
FCS	Thermo Fisher Scientific	A5256701
FBS U2OS	Sigma	F0926-500ML
5 mm diameter coverslips	Thomas Scientific	64–0700
NA 0.71 water dipping illumination objective	Special Optics	54-10-7
25X/NA 1.1 water dipping detection objective	Nikon Instruments	CFI75 Apo LWD 25XY
Camera ORCA-Flash 4.0	Hamamatsu Photonics	C11440-22C
Bidirectional scan unit	Cambridge Technology	6215
Remote focusing system	Nikon Instruments	CFI S Plan Fluor ELWD
Achromatic doublet	Edmund Optics	49–396
Filters	Chroma Technology Corporation	ZET405/488/561/640, ZT568rdc, ET525/50m, ET600/50m
Piezzo Actuator	Physik Instrumente	P-603.1S2
Piezzo controller	Physik Instrumente	E−709.SRG