Prior to the start of the current study, we determined doses of the chemical stimuli that are psychophysically equivalent by eliciting similar numbers of eye wipe responses (Farazifard et al., 2005 (link); Aicher et al., 2015 (link)). The osmolalities of the chemicals used for ocular stimulation were measured on a vapor pressure osmometer (VAPRO Model 5600, Wescor, Inc., Logan, UT). Vehicle control animals received 0.9% saline containing 4% ethanol, 4% Tween-80 (Sigma-Aldrich, St. Louis, MO) (295 mOsm). Chemical stimuli for experimental animals included: 50 mM menthol ((Sigma) in 4% ethanol, 4% Tween-80 in 0.9% saline (298 mOsm)); 33 μM capsaicin ((Sigma) in 0.01% ethanol, 0.01% Tween-80 in 0.9% saline (286 mOsm)); and 5 M hypertonic saline (9430 mOsm).
Vapro model 5600
Manufactured by Wescor
Sourced in United States
The VAPRO Model 5600 is a vapor pressure osmometer designed for measuring the osmolality of a wide range of sample types, including biological fluids, solutions, and other liquid samples. The instrument uses the vapor pressure depression method to determine the osmolality of the sample.
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Lab products found in correlation
5 protocols using vapro model 5600
1
Ocular Stimulation Protocol Optimization
2
Osmotic Pressure Measurement of SOF
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A 10 mL sample of SOF or vitreous fluid was prepared for the osmotic measurement. The osmotic pressure of the sample was measured by a vapor pressure osmometer (Wescor VAPRO Model 5600, Wescor, Logan, UT, USA) following the manufacturer’s instructions.
3
Serelaxin Effects on Murine Metabolic Parameters
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After two weeks of treatment with either placebo or serelaxin, mice were anaesthetised by a cocktail of ketamine (85 mg/kg) and xylazine (8.5 mg/kg) via intraperitoneal injections followed by cardiac puncture for blood collection. Two μL of whole blood was used to measure HbA1c using a cobas b 101 POC system (Roche, Basel, Switzerland). Blood plasma was used to determine plasma osmolality using a vapour pressure osmometer (VAPRO Model 5600, Wescor Inc, Logan, Utah) and concentrations of serelaxin using a Human Relaxin-2 Quantikine ELISA kit (Cat# DRL200; R&D systems, Minneapolis, MN, USA) following manufacturer’s protocol with detection sensitivity of 4.57 pg/mL. Mesenteric arcade, whole heart and aorta were isolated and immediately placed in ice cold Krebs bicarbonate solution (120 mM NaCl, 5 mM KCl, 1.2 mM MgSO4, 1.2 mM KH2PO4, 25 mM NaHCO3, 11.1 mM D-glucose, 2.5 mM CaCl2) and then cleared of fat and connective tissues.
4
Monitoring Cytoplasmic Calcium Dynamics
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The cytoplasmic calcium concentration was monitored by free calcium indicator GCaMP6f (27 (link)). Tmem63b-P2A-GCaMP6f vectors were transfected into N2a cells mounted on the coverslip. GCaMP6f vector was used as a control. 40 h after transfection, the cells were perfused with isotonic extracellular solution (in mm ): 65 NaCl, 5 KCl, 1 CaCl2, 1 MgCl2, and 10 HEPES (pH 7.4 adjusted with NaOH; 300 mOsm/liter adjusted with mannitol). The isotonic solution was exchanged to 170 mOsm/liter hypotonic solution without changing the ionic concentrations by a peristaltic pump (BT100-2J, Longer Precision Pump Co., Ltd., Hebei, China) at a constant speed. The osmolarity was measured by a vapor pressure osmometer (Wescor Vapro Model 5600). The cytoplasmic calcium fluorescence was recorded at 1 Hz for 10 min by laser scanning confocal microscope (Leica Microsystems, TCS SP2) at room temperature (24 ± 2 °C) using 488-nm illumination. The change of fluorescence was normalized by the ratio of real-time intensity (Ft) relative to the initial value (F0). The cells with Ft/F0 > 1.5 were considered as positive cells responding to osmolarity changes.
5
Osmolality Analysis of Diluted Gels
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The osmolality was analyzed using Vapro® Model 5600, (Wescor, Inc., Logan, UT, USA), calibrated with osmolality standards of 100 and 290 mmol/kg. The gels were diluted with DI water to obtain the appropriate consistency in order to completely cover the sample disc. Once analyzed, the measured values were adjusted to the dilution factor and represented as an average of three runs (n = 3).
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