Table 1.
Detailed parametric comparison of the different techniques
| Technique | Wide-field microscopy | Confocal microscopy | Multiphoton microscopy | Light sheet microscopy |
|---|---|---|---|---|
| Standard spatial resolution | < 0.5 µm (lateral) and not applicable (axial) | < 0.4 µm (lateral) and < 2 µm (axial) | < 0.5 µm (lateral) and < 3 µm (axial) | < 0.5 µm (lateral) and < 3 µm (axial) |
| Imaging speed | High (area) | Slow (point scanning) | Slow (point scanning) | High* (volume scan) |
| Imaging depth | Limited | Limited | Best | Limited/high |
| Illumination shape | Wide-field and uniform illumination | Point illumination (hourglass shaped excitation) | Point illumination (nonlinear focal excitation) | Light sheet illumination (decoupled from detection) |
| Detection | Wide-field detector | Point detector | Point detector | Wide-field detector |
| Key application | Thin micro-slice 2D imaging | Very high-volumetric resolution | Deep tissue volume imaging | Fast volume imaging, transparent or optically cleared specimen |
| Main limitation | No axial resolution, photodamage | Imaging speed, imaging depth, and photodamage | Imaging speed and potential nonlinear photodamage | Imaging depth and inhomogeneity of image quality |
*With the use of a faster scanning approach, such as resonant scanning, the speed can reach up to video rate at the cost of a low signal-to-background-noise ratio (SNR)