Microscopy Construction

Video-rate multi-photon microscope

A) Overall layout of the multiphoton imaging system. Components and beam path in red represent the femtosecond laser excitation. Components in green represent the fluorescence detection pathways and image acquisition. Components in yellow represent the transmitted light DIC system for visualizing neurons during establishment of whole-cell patch.

B) Photograph of the microscope system. The laser scanner is mounted in the gray box to the left of the microscope. The locations of the upper and lower photomultiplier (PMT) detectors are indicated, together with the dichroic mirror that reflects laser light down to the objective lens. Paths of the laser beam and of emitted fluorescence light are indicated, respectively, in red and green.

C) Close-up photograph looking down on the scan head, after removing the protective cover. The path of the laser beam is indicated in red.

For more information about the basic design, control circuitry, real-time image corrections,
scan head bracket design, aquisition software, and part list visit

Mike Sanderson's webpage on Real-Time CONFOCAL MICROSCOPY

Nguyen, Q.-T., Callamaras, N., Hseieh, C. & Parker, I. Construction of a two-photon microscope for video-rate Ca2+ imaging. Cell Calcium30(6): 383–393, 2001. [PDF]

1146 Mc Gaugh Hall
University of California, Irvine
Irvine, CA 92697-4550
Lab Tel: 949-824-7833
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last updated 01/17/2006. Please send enquiries to ianparkerlabweb@gmail.com