Multiphoton bio-imaging: how modern femtosecond lasers enable new advancements

Multiphoton microscopy has seen widespread adoption from the research community to address the challenges associated with image collection many hundreds of micrometers into living tissue. Over time, technology advancements have contributed to the evolution of 2-photon, 3-photon and other nonlinear microscopy techniques. Deep Tissue imaging necessitates wavelengths spanning from the visible to the near-IR to achieve desirable resolution. Excitation requires a femtosecond laser pulse to minimize out-of-focus fluorescence. Image collection requires novel broadband microscope objectives which must transmit both the near-IR excitation wavelengths and the subsequent visible fluorescence. During this presentation, a concise overview of the historical development of these technologies will be presented through real-life examples that demonstrate how modern research facilities are leveraging these advancements.