Source: Smail, Y., et al. Imaging Ca2+ Responses During Shigella Infection of Epithelial Cells. J. Vis. Exp. (2018).
This video demonstrates a technique for detecting local and global calcium ion response upon Shigella infection of human epithelial cells. Shigella invades host cells and releases invasion effectors, impacting intracellular calcium levels. The amplitude and frequency of the local and global cytosolic calcium are studied using a fluorescent indicator.
1. Preparations
2. Infection and Image Acquisition of Local Ca2+ Responses
3. Analysis
The authors have nothing to disclose.
Fluo-4 AM | Invitrogen | F14201 | |
Metamorph version 7.7 | Universal Imaging | ||
CoolLED illumination system pE-2 | Roper Scientific | ||
Micro-dish 35 mm, high | IBIDI | 81156 | |
Trypticase Soy (TCS) broth | Thermofisher | B11768 | |
TCS agar | Thermofisher | B11043 | |
Congo red | Sigma-Aldrich | 75768 | |
M90T-AfaE | Sun et al. 2017 | Shigella flexneri serotype V. expressing the AfaE adhesin | |
ipgD-AfaE | Sun et al. 2017 | Isogenic ipgD mutant strain expressing the AfaE adhesin |