Bacterial Infection Quantitation Assay: A Method to Quantify Intraocular Bacterial Load in Mouse Model of Bacterial Endophthalmitis
Published: April 30, 2023
Abstract
Source: Mursalin, M. H. et al., Intravitreal Injection and Quantitation of Infection Parameters in a Mouse Model of Bacterial Endophthalmitis. J. Vis. Exp. (2021).
This video describes the technique of quantifying bacterial infection from eye homogenates obtained from a murine model of bacterial endophthalmitis caused by Bacillus cereus. This technique helps study the host immune response and its dependency on the host and ocular bacterial gene expression in murine models.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Harvesting tube preparation Place 1 mm sterile glass beads into 1.5 mL screw cap tubes. Sterilize these tubes in an autoclave on a dry setting. Let the tubes cool to room temperature before use. Add 10 mL of 1x sterile phosphate-buffered saline (PBS) to a sterile 15 mL centrifuge tube. Add 1 tablet of prote…
Representative Results
Figure 1: Harvesting mouse infected eye. Post-infection, after the desired time point, harvest infected mouse eyes using sterile tweezers. (A) PBS containing sterile glass beads. (B) Harvesting of the infected eye. (C) Harvest tube containing a mouse eye. <img alt=…
Disclosures
The authors have nothing to disclose.
Materials
| 2-20 µL pipette | RANIN | L0696003G | NA |
| 37oC Incubator | Fisher Scientific | 11-690-625D | NA |
| Bacto Brain Heart Infusion | BD | 90003-032 | NA |
| Cell Microinjector | MicroData Instrument, Inc. | PM2000 | NA |
| Fine tip forceps Thermo | Fisher Scientific | 12-000-122 | NA |
| Glass beads 1.0 mm | BioSpec | 11079110 | NA |
| Incubator Shaker | New Brunswick Scientific | NB-I2400 | NA |
| Microcapillary Pipets 5 Microliters | Kimble | 71900-5 | NA |
| Micro-Pipette Beveler | Sutter Instrument Co. | BV-10 | NA |
| Microscope Axiostar Plus | Zeiss | NA | |
| Microscope OPMI Lumera | Zeiss | NA | |
| Mini-Beadbeater-16 | BioSpec | Model 607 | NA |
| Needle/Pipette Puller | Kopf | 730 | NA |
| PBS | GIBCO | 1897315 | Molecular grade |
| Protease Inhibitor Cocktail | Roche | 4693159001 | Molecular grade |
| Reverse action forceps | Katena | K5-8228 | NA |
Tags
Cite This Article
Bacterial Infection Quantitation Assay: A Method to Quantify Intraocular Bacterial Load in Mouse Model of Bacterial Endophthalmitis. J. Vis. Exp. (Pending Publication), e20817, doi: (2023).