Random-Access Multiphoton Microscopy for Fast Three-Dimensional Imaging

  • Gaddum Duemani Reddy
  • Ronald J. Cotton
  • Andreas S. Tolias
  • Peter Saggau
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 859)


Studies in several important areas of neuroscience, including analysis of single neurons as well as neural networks, continue to be limited by currently available experimental tools. By combining molecular probes of cellular function, such as voltage-sensitive or calcium-sensitive dyes, with advanced microscopy techniques such as multiphoton microscopy, experimental neurophysiologists have been able to partially reduce this limitation. These approaches usually provide the needed spatial resolution along with convenient optical sectioning capabilities for isolating regions of interest. However, they often fall short in providing the necessary temporal resolution, primarily due to their restrained laser scanning mechanisms. In this regard, we review a method of laser scanning for multiphoton microscopy that overcomes the temporal limitations of pervious approaches and allows for what is known as 3D Random Access Multiphoton (3D RAMP) microscopy, an imaging technique that supports full three dimensional recording of many sites of interest on physiologically relevant time scales.


Multiphoton Microscopy Three-dimensional Random-access scanning Acousto-optic deflector Spatial dispersion Temporal dispersion Motion tracking di 8 ANEPPS 



The development of AOD-based fast 3D random-access multi-photon microscopy is the result of the continuing efforts of many members and affiliates of the Saggau and Tolias labs. These efforts were supported by numerous grants from NIH and NSF.


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Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • Gaddum Duemani Reddy
    • 1
  • Ronald J. Cotton
    • 1
  • Andreas S. Tolias
    • 1
  • Peter Saggau
    • 1
    • 2
  1. 1.Department of NeuroscienceBaylor College of MedicineHoustonUSA
  2. 2.Allen Institute for Brain ScienceSeattleUSA

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