title: A pragmatic guide to multiphoton microscope design.
abstract: Multiphoton microscopy has emerged as a ubiquitous tool for studying microscopic structure and function across a broad range of disciplines. As such, the intent of this paper is to present a comprehensive resource for the construction and performance evaluation of a multiphoton microscope that will be understandable to the broad range of scientific fields that presently exploit, or wish to begin exploiting, this powerful technology. With this in mind, we have developed a guide to aid in the design of a multiphoton microscope. We discuss source selection, optical management of dispersion, image-relay systems with scan optics, objective-lens selection, single-element light-collection theory, photon-counting detection, image rendering, and finally, an illustrated guide for building an example microscope.
authors.affiliation: Center for Microintegrated Optics for Advanced Biological Control, Department of Physics, Colorado School of Mines, 1500 Illinois Street, Golden, Colorado 80401, USA.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Young
authors.affiliation: W. M. Keck Laboratory for Raman Imaging of Cell-to-Cell Communications, Colorado State University, Fort Collins, Colorado 80523, USA; Department of Electrical and Computer Engineering, Colorado State University, Fort Collins, Colorado 80523, USA.
authors.forename: Jeffrey J
authors.initials: JJ
authors.lastname: Field
authors.affiliation: Photonics Research Center, Department of Physics and Nuclear Engineering, United States Military Academy, West Point, New York 10996, USA.
authors.forename: Kraig E
authors.initials: KE
authors.lastname: Sheetz
authors.affiliation: W. M. Keck Laboratory for Raman Imaging of Cell-to-Cell Communications, Colorado State University, Fort Collins, Colorado 80523, USA; Department of Electrical and Computer Engineering, Colorado State University, Fort Collins, Colorado 80523, USA; School of Biomedical Engineering, Colorado State University, Fort Collins, Colorado 80523, USA.
authors.forename: Randy A
authors.initials: RA
authors.lastname: Bartels
authors.affiliation: Center for Microintegrated Optics for Advanced Biological Control, Department of Physics, Colorado School of Mines, 1500 Illinois Street, Golden, Colorado 80401, USA.
authors.forename: Jeff
authors.initials: J
authors.lastname: Squier