Assessing Bacterial Colonization in a Mouse Brain following Listeria monocytogenes Infection

Abstract

Source: Ghosh, P., et al., Listeria monocytogenes Infection of the Brain. J. Vis. Exp. (2018)

This video demonstrates the quantification of the infective bacterial load from the perfused mouse brain infected with Listeria monocytogenes.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee. 

1. Organ Harvesting and Determination of Bacterial Burdens

1. Following perfusion, harvest organs (e.g., liver, spleen) by carefully removing any vasculature and ligaments attached and placing the organs separately into a sterile 15 mL conical tube containing 5 mL of sterile cold (4 °C) PBS. Store samples on ice until further processing occurs.
2. To harvest the brain, decapitate the head behind the ears using scissors and cut the scalp skin in between the an....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Brain Heart Infusion media	Becton Dickinson	237200
Streptomycin sulfate	Amresco	0382-50G
Petri dishes	VWR	25384-342
Glycerol	VWR	97062-832
IKA T18 ULTRA-TURRAX Basic Homogenizer	IKA	3352109	Model: T18BS1
Spectrophotometer	Beckman Coulter	DU 800 series
BALB/c mice	Jackson Laboratory	Model #000651
Ethylenediaminetetraacetic acid	Sigma-Aldrich	60004
15 ml conical tubes	VWR	21008-918
Phosphate-buffered saline	Corning	46-013-CM
Stainless steel spatula	VWR	82027-520
Stainless steel scissors (4.5 in)	VWR	82027-578
Stainless steel fine tip forceps (6 in)	VWR	82027-406