Multiphoton adaptation of a commercial low-cost confocal microscope for live tissue imaging

• Published in: Journal of Biomedical Optics Vol. 14; no. 3; pp. 034048 - 034045
• Main Authors: Mancuso, James J, Larson, Adam M, Wensel, Theodore G, Saggau, Peter
• Format: Journal Article
• Language: English
• Published: United States 01.05.2009
• Subjects: Animals; Computer programs; Confocal; confocal optics; Detectors; Equipment Design; Femtosecond; fiber coupled detection; Fiber optics; Green Fluorescent Proteins - biosynthesis; Green Fluorescent Proteins - chemistry; Green Fluorescent Proteins - genetics; Humans; Image Processing, Computer-Assisted - instrumentation; Image Processing, Computer-Assisted - methods; instrumentation; Lasers; Mice; Mice, Transgenic; Microscopy; Microscopy, Confocal - instrumentation; Microscopy, Confocal - methods; Microscopy, Fluorescence, Multiphoton - instrumentation; Microscopy, Fluorescence, Multiphoton - methods; multiphoton processes; Neurons; Optical Fibers; Pollen - ultrastructure; pulsed laser; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Retina - cytology; Retina - metabolism; Rhodopsin - biosynthesis; Rhodopsin - chemistry; Rhodopsin - genetics
• ISSN: 1083-3668; 1560-2281
• DOI: 10.1117/1.3139850

The Nikon C1 confocal laser scanning microscope is a relatively inexpensive and user-friendly instrument. We describe a straightforward method to convert the C1 for multiphoton microscopy utilizing direct coupling of a femtosecond near-infrared laser into the scan head and fiber optic transmission of emission light to the three-channel detector box. Our adapted system can be rapidly switched between confocal and multiphoton mode, requires no modification to the original system, and uses only a few custom-made parts. The entire system, including scan mirrors and detector box, remain under the control of the user-friendly Nikon EZ-C1 software without modification.