TPE microscopy broke into the arena bringing its intrinsic three-dimensional resolution, the absence of background fluorescence, and the charming possibility of exciting UV excitable fluorescent molecules at infrared wavelengths increasing sample penetration. Moreover, TPE microscopy can be considered an essential imaging system for thick sample and live cell imaging. At present, the main disadvantages of TPE fluorescence microscopy come from the cost of femtosecond laser sources and the difficulty in predicting or measuring two-photon absorption spectra of the fluorescent molecules potentially utilizable. As also pointed out by other researchers once technology becomes less expensive and simpler, there will not be a confocal, or a light, microscope that is not also a two-photon microscope. TPE applications range from calcium imaging to studies on chromatin structure, from photobleaching characterisation to the utilisation of the two-photon microscope as active nano- micropattering device, from live cell studies to transport processes in nanoenviroments as nanocapsules. Nanosurgery applications and FISH applications are in a period of intense growing. Figure 7 reports an example of nanosurgery from a recent paper by Konig.
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