Cells were grown overnight at 30°C in YPD medium with 80 mg/l uridine, diluted into fresh medium, and incubated again overnight. Cells were washed with phosphate-buffered saline and then diluted so that the desired inoculum could be injected in 0.2 ml. Cell density was determined using a hemocytometer and confirmed by plating dilutions of cells onto YPD agar plates. Female BALB/c mice were injected with the indicated C. albicans strain into the lateral tail vein. The analysis was carried out in two independent experiments in which at least three mice were infected each time. Mice were considered to be moribund if they could no longer reach food and water and were then killed humanely. Fungal burden was quantified by disrupting kidneys for 30 s with a tissue homogenizer (Pro Scientific, Oxford, CT), plating serial dilutions of the homogenate onto YPD plates, and then determining the number of colony-forming units (CFU) per gram of kidney tissue. Samples of homogenized kidney tissue were also pelleted by centrifugation, resuspended in 20% KOH, incubated for 1 h at room temperature, stained with Calcofluor White (20 ng/ml) for 10 min (Pringle, 1991 (link)), and then examined by fluorescence microscopy.
Tissue homogenizer
Manufactured by PRO Scientific
Sourced in United States
A tissue homogenizer is a laboratory instrument used to disrupt and break down solid tissue samples into a fine, homogeneous suspension. It operates by mechanically shearing the tissue through the use of a rotating blade or pestle, allowing for efficient extraction and analysis of cellular components.
Automatically generated - may contain errors
Lab products found in correlation
Genomic lysis buffer, by Zymo Research (3 mentions)
Zymo spin 3 column, by Zymo Research (3 mentions)
Phosphatase inhibitor cocktail, by Merck Group (2 mentions)
Anti gapdh, by Cell Signaling Technology (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Tissue lysis buffer, by Beyotime (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
7 protocols using tissue homogenizer
1
Quantifying Fungal Burden in Murine Candidiasis
2
Extraction and Purification of DNA from Maize Pests
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Specimens were obtained as larvae from corn (maize) plants at various locations in Togo, Africa from July to November 2016 (Fig 1 ). Specimens were stored either air-dried or in ethanol at room temperature. A portion of each specimen was excised and homogenized in 1.5 ml of phosphate buffered saline (PBS, 20 mM sodium phosphate, 150 mM NaCl, pH 8.0) using a tissue homogenizer (PRO Scientific Inc., Oxford, CT, USA) and the homogenate transferred to a 2-ml microcentrifuge tube. The unused portion was stored in ethanol at -20°C. The homogenized tissue was pelleted by centrifugation at 6000 x g for 5 min. at room temperature and the pellet resuspended in 800 μl of Genomic Lysis buffer (Zymo Research, Orange, CA, USA) and incubated at 55°C for 5–30 min. Debris was removed by centrifugation at 10,000 rpm for 3 min. The supernatant was transferred to a Zymo-Spin III column (Zymo Research, Orange, CA, USA) and processed according to manufacturer’s instructions. The DNA preparation was increased to a final volume of 100 μl with distilled water.
3
Genomic DNA Extraction from Fall Armyworm
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Individual specimens were homogenized in 1.5 ml of phosphate buffered saline (PBS, 20 mM sodium phosphate, 150 mM NaCl, pH 8.0) using a tissue homogenizer (PRO Scientific Inc., Oxford, CT, USA) and the homogenate transferred to a 2-ml microcentrifuge tube. Cells and tissue were pelleted by centrifugation at 6000 g for 5 min. at room temperature. The pellet was resuspended in 800 μl Genomic Lysis buffer (Zymo Research, Orange, CA, USA) by vortexing and incubated at 55°C for 5 min. Debris was removed by centrifugation at 10,000 rpm for 3 min. The supernatant was transferred to a Zymo-Spin III column (Zymo Research, Orange, CA, USA) and processed according to manufacturer’s instructions. The DNA preparation was increased to a final volume of 100 μl with distilled water. Genomic DNA preparations of fall armyworm samples from previous studies were stored at -20°C.
4
Genomic DNA Isolation from Tissue
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To isolate DNA, samples were homogenized in 1.5 ml of phosphate buffered saline (PBS, 20 mM sodium phosphate, 150 mM NaCl, pH 8.0) using a tissue homogenizer (PRO Scientific Inc., Oxford, CT) or hand-held Dounce homogenizer then pelleted by centrifugation at 6000g for 5 min. at room temperature. The pellet was resuspended in 800 µl Genomic Lysis buffer (Zymo Research, Orange, CA) and incubated at 55°C for 15 min, followed by centrifugation at 12,000 rpm for 5 min. The supernatant was then transferred to a Zymo-Spin III column (Zymo Research, Orange, CA) to isolate genomic DNA according to the manufacturer’s instructions. Genomic DNA was stored at -20°C until sequencing.
5
Quantification of Matrix Metalloproteinases
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Protein was extracted from indicated tissues in lysis buffer (Thermo Fisher Scientific, Catalog No. 78510) with a protease inhibitor cocktail (Sigma, Catalog No. P8340) and a phosphatase inhibitor cocktail (Sigma, Catalog No. P5726) using a tissue homogenizer (PRO Scientific). Equal amounts of protein were separated by SDS‐PAGE and transferred to a polyvinylidene fluoride membrane. After blocking with 5% BSA buffer, the membrane was probed with anti‐MMP (matrix metalloproteinase) 3 (Abcam, Catalog No. ab52915), anti‐MMP10 (Abcam, Catalog No. ab199688), and anti‐TIMP1 (tissue inhibitor of metalloproteinase 1; Abcam, Catalog No. ab179580) and then developed with chemiluminescent substrate (Thermo Fisher Scientific, Catalog No. 34577). Anti‐GAPDH (Cell Signaling, Catalog No. 2118) was used as internal control to guarantee equal protein loading.
6
Quantifying Intestinal TNF-α Levels After TBI
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The TNF-α contents in the terminal ileum tissue were expressed as nanograms of TNF-α per gram of tissue protein. At 3, 6, or 12h after TBI±labetalol or sham procedure, the levels of intestinal tumor necrosis factor-α (TNF-α) in the tissue supernatant fluids were measured using an ELISA kit (San Diego, CA,USA) specific for rat TNF-α[21 (link),22 (link)]. The minimum limit of TNF-α detection for this assay was 15 pg/mL. The terminal ileum tissue (100 mg per rat) was transferred into a 5 mL tube, and 1mL tissue lysis buffer (Beyotime, Shanghai, China) supplemented with protease inhibitor cocktail (Roche, 04693132001) and phosphatase inhibitor cocktail (Sigma-Aldrich, Germany) was added. The tissue was homogenized on ice using a tissue homogenizer (Pro Scientific, USA); the tissue homogenate was transferred into a Dounce tissue grinder and further processed. The homogenate then was transferred into 1.5 mL tubes and centrifuged for 15 min at 16000g at 4°C; the supernatant fluid was removed and its protein concentration was determined (Pierce Biotechnology, Rockford, USA). The samples were subsequently diluted with deionized water to achieve a concentration of 4 mg protein in 1 mL total volume.
7
Quantifying Fungal Burden in Murine Tongues
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Tongues were excised from mice at the indicated times after infection, weighed, placed in 5 ml PBS, and homogenized for 30 s with a tissue homogenizer (Pro Scientific, Inc., Oxford, CT). The CFU per gram of tongue tissue was then determined by plating serial dilutions of the homogenates onto YPD medium plates and incubating them at 30°C for 2 days.
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