Intravital microscopy

[2] Mice are the most common choice of animals for intravital microscopy but in special cases other rodents such as rats might be more suitable.

The main advantage of intravital microscopy is that it allows imaging living cells while they are in the true environment of a complex multicellular organism.

Another advantage of intravital microscopy is that the experiment can be set up in a way to allow observing changes in a living tissue of an organism over a period of time.

Advancements in fluorescent protein technology and genetic tools that enable controlled expression of a given gene at a specific time in a tissue of interest also played an important role in intravital microscopy development.

The main consideration for the choice of a particular technique is the penetration depth needed to image the area and the amount of cell-cell interaction details required.

If the area of interest is located more than 50–100 μm below the surface or there is a need to capture small-scale interactions between cells, multiphoton microscopy is required.

[6] The maximum depth for the imaging with multiphoton microscopy depends on the optical properties of the tissue and experimental equipment.

More vascularized tissues are generally more difficult to image because red blood cells cause absorption and scattering of the microscope light beam.

In addition, generating transgenic mice with a phenotype of interest and fluorescent proteins in appropriate cell types is often challenging and time-consuming.