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. 2018 Jun 15;5:179. doi: 10.3389/fmed.2018.00179

Table 2.

Description of the most commonly used microscopy for imaging.

BrightField Principle Advantages Drawbacks References
WIDEFIELD MICROSCOPY
Sample directly hit by the white light Easy to use Resolution around 200 nm (118)
OPTICAL IMAGING
Fluorescence Microscopy Sample is illuminated by ultra-violet light to excite the fluorescent dye within the sample;
Separation of emitted light from the excitation light (bright), by specific filters		 Possibility to perform experiments under physiological conditions, making unnecessary chemical fixation and, therefore, minimizing artifacts Risk of photobleaching and quenching related to the exposure time (119, 120)
Bioluminescence Imaging Detection of light emitted from cells, in which enzymes generating light are expressed, by genetic engineering. These enzymes belong to the luciferase group High sensitivity;
Cost- effectiveness;
High reproducibility;
Non-invasive		 Use of genetic engineering tools (luc/neo-MSC);
Low resolution;
Low penetration depth;
Low quantification accuracy;
High light scattering		 (121, 122)
Confocal Microscopy Each spot scanned by the laser, back-and-forth Imaging completed pixel by pixel;
Use of the pinhole;
Final image as an ensemble of layers, much detailed;
Non invasive		 Expensive;
High sensitivity;
Trained operators required		 (119, 123, 124)
Two-Photon Microscopy Use of two low-energy photons, usually from the same laser, cooperating to cause a higher-energy electronic transition in a fluorescent molecule (usually near-infrared light is used) High resolution;
Minimized scattering light;
Increased penetration depth;
Reduced photobleaching
Reduced Background strongly;
Increased total signal-to-noise ratio		 Low penetration depth;
Poor possibility of longitudinal studies		 (125, 126)
LightSheet Microscopy Use of a thin plane of light, instead of a point Very fast imaging speed;
Very high resolution;
Very high penetration depth (>1 cm);
Reduced photobleaching;
Reduced phototoxicity		 Dual side illumination not easy to be perfectly aligned in the three-dimensional space, causing a less sample focus, compared to the single side illumination (127)
MRI Principle Advantages Drawbacks References
MAGNETIC RESONANCE IMAGING
Alignment of the magnetic moment from endogenous molecules (1H and 19F) into an external magnetic field High resolution;
High sensitivity of cell detection;
High penetration depth;		 Low possibility of longitudinal studies;
High costs;
Presence of external metal disturb the analysis		 (110)