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Updated: Aug 11, 2025

Author Spotlight: Enhanced Multiplex Immunofluorescent Microscopy Protocol for Neuroscience Research
Published on: June 21, 2024
Tutorial: multiphoton microscopy to advance neuroscience research
Iván Coto Hernández1, Jenny Yau1, Lars Rishøj2
1Surgical Photonics & Engineering Laboratory, Mass Eye and Ear, Harvard Medical School, 243 Charles St, Boston, MA, United States of America.
Abstract:
Multiphoton microscopy (MPM) employs ultrafast infrared lasers for high-resolution deep three-dimensional imaging of live biological samples. The goal of this tutorial is to provide a practical guide to MPM imaging for novice microscopy developers and life-science users. Principles of MPM, microscope setup, and labeling strategies are discussed. Use of MPM to achieve unprecedented imaging depth of whole mounted explants and intravital imaging via implantable glass windows of the mammalian nervous system is demonstrated.

