Bioluminescence Imaging to Track Bacterial Infection and Wound Healing in a Mouse Model

초록

Source: Hunt, A. M. A., et al. Come to the Light Side: In Vivo Monitoring of Pseudomonas aeruginosa Biofilm Infections in Chronic Wounds in a Diabetic Hairless Murine Model. J. Vis. Exp.(2017).

This video demonstrates the procedure for assessing infection clearance and healing progression in a wounded diabetic mouse using bioluminescent and microscopic images.

프로토콜

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Wound Measurement and Imaging
   1. Evaluate the clinical signs of animals daily.
   2. Provide food, water, and change cages as needed.
   3. Check the integrity of dressings daily. When the dressing is present, only bioluminescence can be measured due to occlusion of the wound. At day 8, dressings are removed and not replaced allowing measurement of wound closure.
   4. Weigh animals every other day.
   5. For all days, place mice individually in an isolation chamber equipped with a HEPA filter and image using the in vivo imaging system daily or every other day until luminescence values fall below the background level.
   6. After day 8, induce general anesthesia and take micrographs of the wound with a microscope with an attached camera using a ruler for reference every other day until wounds are healed.

재료

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이름	회사	카탈로그 번호	댓글
Opsite	Smith & Nephew	Model 66000041	Smith & Nephew Flexfix Opsite Transparent Adhesive Film Roll 4" x 11yards
SKH-1 mice Crl:SKH1-Hrhr	Charles River Breeding Laboratories	SKH1	Hairless mice, 8 weeks old
Streptozotocin (STZ)	Sigma Aldrich	S0130-1G	Streptozocin powder, 1g
AccuChek glucometer	Accu-Chek Roche	Art No. 05046025001	ACCU-CHEK CompactPlus Diabetes Monitoring Care Kit
Pseudomonas aeruginosa Xen 41	Perkin Elmer	119229	Bioluminescent Pseudomonas aeruginosa
Dulbelcco phosphate buffer saline (DPBS)	Sigma Aldrich	D8537	PBS
Tryptic soy agar	Sigma Aldrich	22091	Culture agar
Meloxicam	Henry Schein Animal Health	49755	Eloxiject (Meloxicam) 5mg/mL, solution for injection
10% povidone-iodine (Betadine)	Purdue Products LP	301879-OA	Swabstick, Betadine Solution. Antiseptic. Individ. Wrapped, 200/case
Silicone to make splints	Invitrogen Life Technologies Corp	P-18178	Press-to-Seal Silicone Sheet, 13cm x 18cm, 0.5mm thick, set of 5 sheets
HEPA filter	Teleflex	28022	Gibeck ISO-Gard HEPA Light number 28022
Biopsy punches	VWR International Inc	21909-142	Disposable Biopsy Punch, 5mm, Sterile, pack of 50.
Biopsy punches	VWR International Inc	21909-140	Disposable Biopsy Punch, 4mm, Sterile, pack of 50.
Glucose	J.T.Baker	1916-01	Dextrose, Anhydrous, Powder
Corning 96-well black plates	Fisher Scientific	07-200-567	96-well clear bottom black polysterene microplates
25 gauge 5/8 inch needle	BD	305122	Regular bevel needle