Table 2.

Classification of imaging infrastructure by “ease of use”, “maintenance”, “data size”, “quantifiable data output”, and “post-processing requirements”. “Simple” instruments can be operated after one day of training. “Advanced” instruments have more complex modalities and parameter settings, and require additional training. “Expert” instruments require multiple weeks of training, including experimental set up, appropriate controls and calibrations. Star (*) denotes that the user-friendliness is strongly dependent on the particular setup. “Maintenance” estimates whether the standard procedures including hardware management can be carried out by a trained user, a specialist, or an engineer, for example in a dedicated imaging facility. “Data size” refers to image files of a typical experiment, ranging from “small” (megabyte range) to “very large” (hundreds of gigabytes). Note that time course or high-throughput experiments significantly increase data size. “Quantifiable data output” denotes if a linear ratio of fluorescence excitation to emission is obtained, which can be used for intensity comparisons between different experimental conditions. “Post-processing” indicates whether images can be directly used for analysis, or if they require additional steps such as alignment, averaging or reconstruction.

	Ease of Use	Maintenance	File Sizes	Quantifiable	Postprocessing
Widefield	Simple	User	Small	Yes	No
CLSM	Simple	Specialist	Moderate	Yes	No
SDCM	Simple	Specialist	Moderate	Yes	No
2-photon	Expert	Engineer	Moderate	Yes	No
Airyscan	Simple	Engineer	Moderate	Yes	Yes
Lightsheet	Advanced	Specialist	Very large	Yes	Yes
STED	Advanced	Specialist	Small	Yes	No
PALM/STORM	Simple–Expert *	Specialist	Very large	No	Yes
SIM	Simple–Expert *	Engineer	Moderate	No	Yes
iSIM	Simple	Engineer	Moderate	Yes	No
DHM	Simple	User	Large	Yes	Yes

Abbreviations: CLSM—Confocal Laser Scanning Microscope, DHM—Digital Holographic Microscope, iSIM—instant Structured Illumination Microscopy, PALM—Photo-Activated Localization Microscopy, SDCM—Spinning Disk Confocal Microscope, SIM—Structured Illumination Microscopy, STED—Stimulated Emission Depletion, STORM—Stochastic Optical Reconstruction Microscopy.