Detecting Bacterial Contamination Using Magneto-fluorescent Nanosensors
Published: March 29, 2024
Abstract
Source: Shelby, T., Sulthana, S., McAfee, J., Banerjee, T., Santra, S. Foodborne Pathogen Screening Using Magneto-fluorescent Nanosensor: Rapid Detection of E. Coli O157:H7. J. Vis. Exp. (2017).
This video demonstrates a technique to detect bacterial contaminants in food and water samples using nanosensors. The target pathogenic bacteria are detected through a combination of magnetic relaxation and fluorescence emission modalities.
Protocol
1. Rapid Detection of E. coli O157:H7 using MFnS Spike various PBS solutions (1X, pH 7.4, 300 µL) with increasing amounts of the 10-6 bacterial stock, resulting in CFU ranges from 1-100. Add a consistent amount of MFnS (100 µL) to these solutions. Create one baseline solution that contains only PBS (1X, pH 7.4, 300 µL) and MFnS (100 µL). Incubate solutions for 30 min at 37 °C and then allow them to cool to room temperature….
開示
The authors have nothing to disclose.
Materials
| Ferrous Chloride Tetrahydrate | Fisher Scientific | I90-500 | |
| Ferric Chloride Hexahydrate | Fisher Scientific | I88-500 | |
| Ammonium Hydroxide | Fisher Scientific | A669S-500 | |
| Hydrochloric Acid | Fisher Scientific | A144S-500 | |
| Polyacryllic Acid | Sigma-Aldrich | 323667-100G | |
| EDC | Thermofisher Scientific | 22980 | |
| NHS | Fisher Scientific | AC157270250 | |
| Anti-E. coli O111 antibody | Sera care | 5310-0352 | |
| Anti-E. coli O157:H7 antibody [P3C6 | Abcam | ab75244 | |
| DiI Stain | Fisher Scientific | D282 | |
| Nutrient Broth | Difco | 233000 | |
| Freeze-dried E. coli O157:H7 pellet | ATCC | 700728 | |
| Magnetic Relaxomteter | Bruker | mq20 | |
| Zetasizer | Malvern | NANO-ZS90 | |
| Plate Reader | Tecan | Infinite M200 PRO | |
| Magnetic Column | QuadroMACS | 130-090-976 | |
| Centrifuge | Eppendorf | 5804 Series | |
| Centrifuge (accuSpin Micro 17) | Fisher Scientific | 13-100-676 | |
| Floor Model Shaking Incubator | SHEL LAB | SSI5 | |
| Analytical Balance | Metler Toledo | ME104E | |
| Digital Vortex Mixer | Fisher Scientific | 02-215-370 | |
| Open-Air Rocking Shaker | Fisher Scientific | 02-217-765 |
