TriMScope - Multiphoton Microscopy

The TriMScope system by LaVison, Göttingen, Germany is the multiphoton microscopy device of the medical faculty of the CAU. Confocal microscopic images can be acquired using illumination by a laser. Unlike conventional laser scanning microscopy two synchronized photons with twice the wavelength (710-980nm) are used to initiate excitation of lower wavelength fluorophores (down to approximately half the excitation wavelength) in a specific plane of imaging. The advantages are lower photo-toxicity effects on living tissue and deeper penetration depth due to lower absorption at longer wavelengths in the so-called optical window of biological material.
In a special mode of operation, Second Harmonic Generation (SHG) imaging can be performed. SHG imaging is used to image non-centrosymmetric protein structures, e.g. to detect organized collagen structures. Like in the case of multiphoton microscopy synchronized higher wavelength excitation photons are transformed to an emission wavelength at exactly half the excitation wavelength. Combination of both techniques allows to synchonously monitor fluorophores and unstained protein structures at high resolution (about 1 µm), e.g. bone marrow stroma (red) and collagen (green), as shown in the figure.