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.
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
2. Harvesting the eyes
3. Intraocular bacterial count
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.
Figure 2: Processing the harvested eye for intraocular bacterial count. (A) Harvest tube with infected eye clamped tightly to a tissue homogenizer. (B) Infected eyes were homogenized twice for 1 min each. Track dilution (D) of the eye homogenates (C) and subsequent plating (E) for the bacterial quantitation. (E) Representative individual Bacillus cereus colony after track dilution.
The authors have nothing to disclose.
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 |