Detection of Bacterial Adherence to Host Cells Using Fluorescence Imaging
Published: October 31, 2023
Abstract
Source: Yang, J., et al. Automated, High-Throughput Detection of Bacterial Adherence to Host Cells. J. Vis. Exp. (2021)
This video demonstrates the bacterial adherence assay to evaluate GFP-tagged bacterial adherence to epithelial cell culture to study the dynamics of host-pathogen interactions using fluorescence microscopy-based imaging.
Protocol
1. Bacterial adherence and host cell staining First, wash the seeded A549 cell monolayers three times with warm 1x PBS. For each wash, add 100 μL of 1x PBS to each well, gently pipette up and down three times, dispose of 1x PBS or wait for 10 s after addition, and then vacuum to remove 1x PBS. To determine the kinetics of bacterial association, overlay the cells with 100 µL of desired concentrations of bacterial suspension with different MOIs (0, 1, 10, and 100). Spin down the bacte…
Divulgations
The authors have nothing to disclose.
Materials
| 10x PBS | VWR | 45001-130 | |
| 4′,6-diamidino-2-phenylindole (DAPI) | Thermo Fisher | 62248 | Host cell staining dye |
| 96-well plate | Corning | 3882 | Half area well, flat clear bottom |
| A549 cells | ATCC | CCL 185 | Mammalian cell line |
| Centrifuge | Eppendorf | 5810R | |
| OD600 DiluPhotometer | IMPLEN | ||
| Typical soy broth | Growcells | MBPE-4040 | |
| F-12k medium | ATCC | 302004 | A549 cell culture medium |
| Fetal bovine serum | Corning | 35-016-CV |
Tags
Citer Cet Article
Detection of Bacterial Adherence to Host Cells Using Fluorescence Imaging. J. Vis. Exp. (Pending Publication), e21663, doi: (2023).