The DEX concentration in the cochlear perilymph were assessed at 30 minutes, 90 minutes, 3 hours, 1 day, 3 days, and 7 days after IT administration. The sampling and analytical methods were carried out in the same manner as previously reported.26 (link) Small cochleostomy was performed at the tip of the cochlea, and then the capillary tubes (Sigma-Aldrich, St. Louis, MO, USA) were used to collect the perilymph. These operations were performed under a surgical microscope. Finally, all the perilymph and standard samples were diluted and analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) method equipped with an Agilent 1290 Infinity II LC system (Agilent Technologies, Santa Clara, CA, USA) and a QTRAP ® 6500 LC-MS/MS System (SCIEX, Framingham, MA, USA). The date scan was performed using multiple reaction monitoring.
Capillary tube
Manufactured by Merck Group
Sourced in United States
A capillary tube is a small, narrow glass or plastic tube used to hold and transport small volumes of liquid. It functions by utilizing capillary action to draw and hold liquids within the tube.
Automatically generated - may contain errors
Lab products found in correlation
Agilent 1290 infinity 2 lc system, by Agilent Technologies (1 mentions)
Qtrap 6500 lc ms ms system, by AB Sciex (1 mentions)
Octyltrichlorosilane, by Merck Group (1 mentions)
Ultrapure water, by Merck Group (1 mentions)
Methylene dichloride, by Merck Group (1 mentions)
Milli q system, by Merck Group (1 mentions)
Chloroform, by Merck Group (1 mentions)
Ethanol, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Sulfuric acid, by Merck Group (1 mentions)
Microcentrifuge tube, by Eppendorf (1 mentions)
5 protocols using capillary tube
1
Perilymph DEX Concentration Measurement
2
Preparation of Lipid Bilayer Membranes
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Tris buffered saline (TBS; 0.05 M Tris, 0.138 M NaCl, and 0.0027 M KCl, pH 8.0), sulfuric acid, H2O2 (30% w/v), octyltrichlorosilane (OTS), 2-arachidonoyl-1-palmitoyl-snglycero-3-phosphocholine (PAPC), AH, sodium chloride, hydrochloric acid, sodium hydroxide, chloroform, ethanol, methanol, methylene dichloride, and capillary tubes were purchased from Sigma-Aldrich (St. Louis, MO). LC, 4′-pentyl-4-cyanobiphenyl (5CB), was purchased from EM Industries (Hawthorne, NY). Copper specimen grids (50 meshes, 500 μm pitch, 420 μm hole, 80 μm bar, 25 ± 5 μm thickness) were obtained from GILDER GRIDS (Grantham, Lincs). Premium glass microscope slides were purchased from Fisher Scientific (Pittsburgh, PA).
Ultrapure water, with a resistivity of 18.2 MΩ cm, was obtained from a Milli-Q system (Millipore, Bedford, MA).
Ultrapure water, with a resistivity of 18.2 MΩ cm, was obtained from a Milli-Q system (Millipore, Bedford, MA).
3
Measuring Fly Food Intake using CAFE Assay
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Food intake was measured using a slightly modified CAFE assay following Ja et al. (39 (link)). In brief, female flies were placed into 1.5-ml Eppendorf microcentrifuge tubes with an inserted capillary tube (5 μl, Sigma-Aldrich) containing 5% sucrose, 2% yeast extract, and 0.1% propionic acid. To estimate evaporation, three food-filled capillaries were inserted in identical tubes without flies. The final food intake was determined by calculating the decrease in food level minus the average decrease in the three control capillaries. Food consumption was measured daily and calculated cumulatively over 4 consecutive days. For this assay, we used 8–10 flies in each of three biological replicates.
4
Quantifying Fly Feeding and Hydration
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This method was modified from Ja et al., 2007 [36] . Individual flies were carefully placed in 1.5 mL Eppendorf microcentrifuge tubes, each equipped with an inserted capillary tube (5 µL, Sigma). To assess feed intake, the capillary tube was filled with a liquid diet consisting of 5% sucrose, 2% yeast extract, and 0.1% propionic acid. To assess water consumption in relation to food, the flies were provided with a capillary tube filled with Milli-Q water in addition to the capillary tube containing food. As controls to account for evaporation, Eppendorf tubes with food and water capillaries but no flies were included.
The final measurement of food intake or water consumption was calculated by subtracting the decrease in food or water levels from the mean decrease in the control capillaries. Food consumption was monitored daily and calculated cumulatively for four consecutive days, using 8 to 10 three-day-old adult male flies for each of the three biological replicates. The entire experimental setup was maintained at a constant temperature of 25 • C in a 12:12 h light/dark cycle.
The final measurement of food intake or water consumption was calculated by subtracting the decrease in food or water levels from the mean decrease in the control capillaries. Food consumption was monitored daily and calculated cumulatively for four consecutive days, using 8 to 10 three-day-old adult male flies for each of the three biological replicates. The entire experimental setup was maintained at a constant temperature of 25 • C in a 12:12 h light/dark cycle.
5
Fly Food and Water Consumption Assay
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The CAFE assay (Ja et al., 2007) was used for measuring food intake and water consumption. Single female flies were placed in 1.5-ml Eppendorf micro centrifuge tubes with an inserted capillary tube (5 µl, Sigma). For the food intake assay, the capillary tube was filled with liquid food containing 5% sucrose, 2% yeast extract and 0.1% propionic acid. This diet, thus, differs from the one used for HFD. To test water consumption relative to feeding, flies were given a capillary tube filled with milli-Q water, in addition to the capillary tube containing food. Eppendorf tubes with food and water capillaries, but without flies, were used as controls for evaporation. The final food intake or water consumption was determined by calculating the decrease in food or water levels minus the average decrease in the control capillaries.
Food consumption was measured daily and calculated cumulatively over four consecutive days. We used 8-10 flies in each of three biological replicates.
Food consumption was measured daily and calculated cumulatively over four consecutive days. We used 8-10 flies in each of three biological replicates.
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