Related Experiment Videos
[Three-dimensional morphology of C6/36 cells infected by dengue virus: a study based on digital holographic
Jian-Hai Yu1, Xu-Ling Liu, Yu-Jing Liu
1Biosafety Level 3 Laboratory, School of Public Health, Southern Medical University, Guangzhou 510515, China.E-mail: chienhai@163.com.
Insights
Digital holographic microscopy reveals distinct 3D morphological changes in C6/36 cells infected with different dengue virus (DENV) serotypes, aiding in understanding DENV infection mechanisms.
Area of Science:
- Cell biology
- Virology
- Microscopy
Background:
- Dengue virus (DENV) infection poses a significant global health challenge.
- Understanding the intricate mechanisms of DENV infection at the cellular level is crucial for developing effective interventions.
- Current methods may not fully capture the dynamic 3D morphological alterations induced by DENV.
Purpose of the Study:
- To monitor the 3D morphological changes of C6/36 cells during DENV infection.
- To utilize live-cell imaging with digital holographic microscopy for this monitoring.
- To gain insights into the mechanisms of DENV infection.
Main Methods:
- C6/36 cells were cultured and optimized for imaging density.
- Cells were subjected to temperature changes (28°C to 37°C).
- Cells were infected with four different DENV serotypes and monitored using HoloMonitor M4 for 3D holograms and morphological parameters.
Main Results:
- Optimal cell density (4×10^5/well) yielded consistent 3D cell morphologies.
- Incubation at 37°C for 24h did not significantly alter uninfected cell morphology.
- DENV infection increased cell area and volume while decreasing thickness; specific serotypes (DENV-1, DENV-2) reduced thickness, while others (DENV-3, DENV-4) increased it.
Conclusions:
- Digital holographic microscopy is effective for monitoring complex cellular changes during DENV infection.
- Different DENV serotypes induce characteristic and distinct cellular damages.
- This technique provides valuable data for understanding DENV pathogenesis.
Objective:
To monitor the 3-dimensional (3D) morphological changes of C6/36 cells during dengue virus (DENV) infection using a live-cell imaging technique based on digital holographic microscopy and provide clues for better understanding the mechanisms of DENV infection.
Methods:
C6/36 cells were seeded in 6-well plates to determine the optimal imaging density under a holographic cell imager, and the morphological changes of the cells were recorded in response to a culture temperature change from 28 degrees celsius; to 37 degrees celsius; C6/36 cells were infected with 4 DENV strains with different serotypes at 28 degrees celsius; and incubated at 37 degrees celsius; for 24 h, and the 3D holograms and relevant morphological parameters were recorded at different time points using HoloMonitor M4 holographic cell imaging and analysis system.
Results:
The holograms of C6/36 cells inoculated at the optimal density for imaging (4×105 per well) showed unified 3D morphologies of the single cells with minimal dispersions in the cell area, thickness and volume (P<0.05), which did not undergo obvious changes when the cells were incubated at 37 degrees celsius; for 24 h (P>0.05). The cell area and volume of the cells infected with the 4 DENV strains all increased and the cell thickness was reduced during incubation. Among the 4 strains, DENV-1 and DENV-2 caused reduced cell thickness while DENV-3 and DENV-4 increased the cell thickness, and the pattern and degree of such changes differ among the 4 strains.
Conclusions:
Digital holographic microscopy allows monitoring of the complex morphological changes of cells during DENV infection. The 4 DENV strains with different serotypes causes characteristic cell damages during infection.