Table 1. Comparison of the available techniques to investigate immune signaling processes (fps: frame per second).

Technique	XY res (nm)	Z res (nm)	Temporal (fps)	Limitation	Advantages	Typical use
Total internal reflection fluorescence (TIRF) microscopy	250	100	100	Only the interface with coverslip	High contrast	Synapse formation, single-molecule dynamics
Variable angle (VA)-TIRF microscopy	250	10	100	Only the interface with coverslip	Z resolution++	Cell topography
Confocal	250	500	20	Photobleaching	Simplicity	Cell structure, interactions
Airyscan	125	250	5	Photobleaching	Resolution	Cell structure, interactions
Stimulated emission depletion (STED) microscopy, ground state depletion (GSD) microscopy	20	50	1	Toxicity, Adequate dyes	Instant super-resolution	Live cell super-resolution, Combination with single molecule
Instant structural illumination microscopy (SIM)	125	250	100	Photobleaching	Speed, resolution	Dynamic cell–cell interactions
Lattice Light-sheet microscopy	250	500	100	Setup complexity	Gentle and fast imaging	Dynamic cell–cell interactions
Lattice SIM	125	250	20	Setup complexity	Gentle and high resolution imaging	Dynamic cell–cell interactions
Single molecular localization microscopy (SMLM)	15	15	-	Chemically fixed cells, Elaborate data analysis	Axial and lateral resolution	Local and global distribution of molecules
Minimal photon fluxes (MINFLUX) microscopy	2 (20)	2 (20)	- (100 µs)	Chemically fixed cells, Elaborate data analysis	Axial and lateral resolution	Cell structure, interactions, Localization of molecules (tracking)