B6FVB TRAMP (+) mice were injected intravenously (tail vein injections) with 200 μl sterile PBS containing 1.0 × 107 or 2.5 × 107 CRC2631iRFP720-cat. Mice were euthanized 190 hours post injection. Whole blood, lung, liver, spleen, kidneys, prostate, and proper axial lymph nodes as well as any discrete metastatic tumor masses were collected, weighed, and kept on ice. Whole blood samples were immediately diluted 1/10 in 25% glycerol and PBS and stored at −80°C. Tissue samples were homogenized in 3 mL sterile PBS for 20 seconds on ice using a TissueRuptor homogenizer (Qiagen) with sterile tips, mixed with 3 mL of sterile 50% glycerol, and stored at −80°C. All tissue samples were later thawed, passed through 40 μm sterile filters (BD Biosciences) and immediately diluted, spotted in triplicate on selective LB +200 μg/ml thymine +20 μg/mL chloramphenicol plates, incubated at 37°C and enumerated after 24 h following the Miles and Misra method [50 (link)].
Sterile filters
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Sterile filters are laboratory devices designed to remove microorganisms and other contaminants from liquids or gases. They are used to maintain sterility and purity in various applications, such as pharmaceutical, biotechnology, and food and beverage production processes.
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2 protocols using sterile filters
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Quantifying Metastatic Tumor Burden in Mice
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Quantifying Metastatic Tumor Burden
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B6FVB TRAMP(+) mice were injected intravenously (tail vein injections) with 200 µl sterile PBS containing 1.0x107 or 2.5x107 CRC2631iRFP720-cat. Mice were euthanized 190 hours post injection.
Whole blood, lung, liver, spleen, kidneys, prostate, and proper axial lymph nodes as well as any discrete metastatic tumor masses were collected, weighed, and kept on ice. Whole blood samples were immediately diluted 1/10 in 25% glycerol and PBS and stored at -80 °C. Tissue samples were homogenized in 3 mL sterile PBS for 20 seconds on ice using a TissueRuptor homogenizer (Qiagen) with sterile tips, mixed with 3 mL of sterile 50% glycerol, and stored at -80 °C. All tissue samples were later thawed, passed through 40 µm sterile filters (BD Biosciences) and immediately diluted, spotted in triplicate on selective LB +200 µg/ml thymine +20 µg/mL chloramphenicol plates, incubated at 37 °C and enumerated after 24 h following the Miles and Misra method [50] .
Whole blood, lung, liver, spleen, kidneys, prostate, and proper axial lymph nodes as well as any discrete metastatic tumor masses were collected, weighed, and kept on ice. Whole blood samples were immediately diluted 1/10 in 25% glycerol and PBS and stored at -80 °C. Tissue samples were homogenized in 3 mL sterile PBS for 20 seconds on ice using a TissueRuptor homogenizer (Qiagen) with sterile tips, mixed with 3 mL of sterile 50% glycerol, and stored at -80 °C. All tissue samples were later thawed, passed through 40 µm sterile filters (BD Biosciences) and immediately diluted, spotted in triplicate on selective LB +200 µg/ml thymine +20 µg/mL chloramphenicol plates, incubated at 37 °C and enumerated after 24 h following the Miles and Misra method [50] .
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