The same cohort of mice was euthanized 1 day after the 6-week antibiotic course. To avoid disturbing the biofilm at the distal end of the implant, the surrounding bone was gently crushed circumferentially with a needle driver and the implant was readily extracted. Bacteria were isolated from the bone and joint tissue by homogenizing tissue from the midpart of the femur through the proximal part of the tibia using a PRO200 Series homogenizer (PRO Scientific). Bacteria adherent to the Kirschner wire implants were isolated by sonication in 0.3% Tween solution (Sigma-Aldrich) for 10 minutes followed by vortexing for 2 minutes. The CFUs were counted after overnight culture with serial dilutions on plates. To further identify any bacteria remaining in the bone or joint tissue or the implants, homogenates and sonicates were cultured for an additional 48 hours (at 37°C in tryptic soy broth at 240 rpm) and the presence or absence of bacterial growth was determined after overnight culture on plates.
Pro200 series
Manufactured by PRO Scientific
Sourced in United States
The Pro200 Series is a line of high-performance laboratory equipment designed for a variety of scientific applications. The core function of the Pro200 Series is to provide precise and reliable measurements, with a focus on accuracy and consistency. The specific capabilities and features of the Pro200 Series may vary depending on the exact model, but the series is generally known for its durability, ease of use, and versatility in the laboratory setting.
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Lab products found in correlation
6 protocols using pro200 series
1
Isolating Bacteria from Implant, Bone, and Tissue
2
Quantifying Bacterial Burden in Implant-Associated Tissues
3
Assessing Implant Infection in Mice
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The experiment was arbitrarily ended on day 28 after the intravenous inoculation. All mice were euthanized, and the bone/joint tissues from the op and contralateral non-op legs as well as the implants were harvested. The bone/joint tissues from each of the op and non-op legs were homogenized (PRO200 Series; PRO Scientific, Oxford, CT, USA), and the tissue homogenates were then serially diluted and cultured overnight on plates at 37 °C. The implants from each of the op legs were sonicated in a 0.3% Tween (Sigma-Aldrich, St. Louis, MO, USA) solution for 10 min, vortexed for 2 min, and then serially diluted and cultured overnight on plates at 37 °C. To further distinguish beTween the presence and absence of an infection, the original tissue homogenates and implant sonicates were incubated in TSB for 48 h at 37 °C with shaking (240 rpm), followed by overnight plating of the broth at 37 °C. Growth in broth and plates were confirmed with BLI of the plates to evaluate for any contamination.
4
Quantifying Bacterial Load in Skin Lesions
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Mice (n = 5/group) were euthanized on day 3 post-infection and 10-mm skin punch biopsies of lesions were homogenized (Pro200 Series homogenizer; Pro Scientific, Oxford, CT) in PBS on ice. Samples were serially-diluted and cultured on TSA plates overnight and CFU were enumerated.
5
Skin Biopsy Homogenization and Ex Vivo CFU
6
Rabbit Model of Staphylococcus Infection
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Two weeks after the last immunization, the IM infection was performed on the right thigh. Rabbits were anesthetized with IM ketamine (20-30 mg/kg) and xylazine (1-2 mg/kg) and general anesthesia was maintained with inhalation isoflurane (∼1.5%). Sterile ophthalmic ointment (Rugby, Livonia, MI, USA) was applied to the eyes. Metoclopramide (Teva) (0.3 mg/kg) was injected subcutaneously as a gastrointestinal promotility agent and sustained-release buprenorphine (ZooPharm) (0.2 mg/kg) was injected subcutaneously for analgesia. For infection, 6-8 ×109 CFU of bioluminescent SF8300 in 100 µl of PBS was injected into the muscle located in the middle of femur with depth of 0.5 cm using a 29-gauge insulin syringe. In vivo BLI was performed using a Lumina III IVIS (PerkinElmer) and maximum flux (photons/s/cm2/steradian) was measured within an 8.1×6.4 cm circular region of interest using Living Image software (PerkinElmer). Thigh width was measured by Pittsburgh digital caliper (Harbor Freight Tools). Ex vivo CFU was enumerated from overnight cultures of serially diluted muscle abscess specimen homogenized at 4°C (Pro200 Series homogenizer; Pro Scientific).
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