This monograph focuses on modern femtosecond laser microscopes for two photon imaging and nanoprocessing, on laser tweezers for cell micromanipulation as well as on fluorescence lifetime imaging (FLIM) in Life Sciences. The book starts with an introduction by Dr. Wolfgang Kaiser, pioneer of nonlinear optics and ends with the chapter on clinical multiphoton tomography, the novel high resolution imaging technique. It includes a foreword by the nonlinear microscopy expert Dr. Colin Sheppard.

Contents
Part I: Basics

Brief history of fluorescence lifetime imaging

The long journey to the laser and its use for nonlinear optics

Advanced TCSPC-FLIM techniques

Ultrafast lasers in biophotonics

Part II: Modern nonlinear microscopy of live cells

STED microscopy: exploring fluorescence lifetime gradients for super-resolution at reduced illumination intensities

Principles and applications of temporal-focusing wide-field two-photon microscopy

FLIM-FRET microscopy

TCSPC FLIM and PLIM for metabolic imaging and oxygen sensing

Laser tweezers are sources of two-photon effects

Metabolic shifts in cell proliferation and differentiation

Femtosecond laser nanoprocessing

Cryomultiphoton imaging

Part III: Nonlinear tissue imaging

Multiphoton Tomography (MPT)

Clinical multimodal CARS imaging

In vivo multiphoton microscopy of human skin

Two-photon microscopy and fluorescence lifetime imaging of the cornea

Multiscale correlative imaging of the brain

Revealing interaction of dyes and nanomaterials by multiphoton imaging

Multiphoton FLIM in cosmetic clinical research

Multiphoton microscopy and fluorescence lifetime imaging for resection guidance in malignant glioma surgery

Non-invasive single-photon and multi-photon imaging of stem cells and cancer cells in mouse models

Bedside assessment of multiphoton tomography