3D Micropatterned Multiphoton Stimulation via Deep Learning-Based Computer-Generated Holography with Temporal Focusing Confinement

Liang Wei Chen, Feng Chun Hsu, Yvonne Yuling Hu, Chun Yu Lin, Shean Jen Chen*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

1 Scopus citations


The customized 3D illumination patterns can be generated with computer-generated holography (CGH), and the axial confinement of the illumination patterns can be improved by inducing the temporal focusing technique. Through these approaches, the neuron excitation in single-cell resolution can be achieved. However, due to the computation cost of iterative CGH algorithm, the hologram must be pre-calculated to generate the illumination patterns for neuron excitation. This shortcoming makes it difficult to dynamically stimulate the neurons for observing neural activity. To overcome this issue for real-time dynamic neuron stimulation, we develop a neuron stimulation system with single-cell resolution and a real-time CGH algorithm. For single-cell resolution, a diffraction grating is used to generate the temporal focusing effect. Moreover, we design a deep-learning based CGH algorithm considering temporal focusing effect to real-time generate hologram with the pre-trained U-net architecture for producing customized illumination patterns in 3D positions. In our approach, the dynamic 3D micro-patterned single-cell neural excitation can be achieved by inducing temporal focusing technique to improve the axial resolution to few microns level and generating hologram by deep-learning based CGH considering temporal focusing to speed up the computation time to tens of milliseconds.

Original languageEnglish
Title of host publicationBiomedical Spectroscopy, Microscopy, and Imaging II
EditorsJurgen Popp, Csilla Gergely
ISBN (Electronic)9781510651647
StatePublished - 2022
EventBiomedical Spectroscopy, Microscopy, and Imaging II 2022 - Virtual, Online
Duration: 9 May 202220 May 2022

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
ISSN (Print)0277-786X
ISSN (Electronic)1996-756X


ConferenceBiomedical Spectroscopy, Microscopy, and Imaging II 2022
CityVirtual, Online


  • computer-generated hologram
  • deep learning
  • multiphoton excitation
  • spatial light modulator
  • temporal focusing


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