The applications of lattice light-sheet microscopy for functional volumetric imaging of hippocampal neurons in a three-dimensional culture system

Chin Yi Chen, Yen Ting Liu, Chieh Han Lu, Po Yi Lee, Yun Chi Tsai, Jyun Sian Wu, Peilin Chen, Bi Chang Chen*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

The characterization of individual cells in three-dimensions (3D)with very high spatiotemporal resolution is crucial for the development of organs-on-chips, in which 3D cell cultures are integrated with microfluidic systems. In this study, we report the applications of lattice light-sheet microscopy (LLSM) for monitoring neuronal activity in three-dimensional cell culture. We first established a 3D environment for culturing primary hippocampal neurons by applying a scaffold-based 3D tissue engineering technique. Fully differentiated and mature hippocampal neurons were observed in our system. With LLSM, we were able to monitor the behavior of individual cells in a 3D cell culture, which was very difficult under a conventional microscope due to strong light scattering from thick samples. We demonstrated that our system could study the membrane voltage and intracellular calcium dynamics at subcellular resolution in 3D under both chemical and electrical stimulation. From the volumetric images, it was found that the voltage indicators mainly resided in the cytosol instead of the membrane, which cannot be distinguished using conventional microscopy. Neuronal volumetric images were sheet scanned along the axial direction and recorded at a laser exposure of 6 ms, which covered an area up to 4800 μm2, with an image pixel size of 0.102 μm. When we analyzed the time-lapse volumetric images, we could quantify the voltage responses in different neurites in 3D extensions.

Original languageEnglish
Article number599
JournalMicromachines
Volume10
Issue number9
DOIs
Publication statusPublished - 2019 Sept 1
Externally publishedYes

Keywords

  • 3D cell culture system
  • Functional neuron imaging
  • Lattice light-sheet microscopy

ASJC Scopus subject areas

  • Control and Systems Engineering
  • Mechanical Engineering
  • Electrical and Electronic Engineering

Fingerprint

Dive into the research topics of 'The applications of lattice light-sheet microscopy for functional volumetric imaging of hippocampal neurons in a three-dimensional culture system'. Together they form a unique fingerprint.

Cite this