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.
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 DENVinfection. 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 DENVinfection. The 4 DENV strains with different serotypes causes characteristic cell damages during infection.
Authors: María Yazi Mendoza; Juan Santiago Salas-Benito; Humberto Lanz-Mendoza; Salvador Hernández-Martínez; Rosa M del Angel Journal: Am J Trop Med Hyg Date: 2002-07 Impact factor: 2.345
Authors: Tanay M Desai; Mariana Marin; Christopher R Chin; George Savidis; Abraham L Brass; Gregory B Melikyan Journal: PLoS Pathog Date: 2014-04-03 Impact factor: 6.823