Model 3320

Manufactured by Advanced Instruments

Sourced in United States, Singapore

The Model 3320 is a laboratory instrument that measures the osmolality of samples. It determines the osmotic concentration of a solution by measuring the freezing point depression. The instrument is designed to provide accurate and reliable osmolality measurements for various applications in research and clinical settings.

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Close spelling variants of this product

Model 3320 Osmometer (10 citations) Advanced® Model 3320 Micro-Osmometer (7 citations) Micro-osmometer Model 3320 (6 citations)

26 protocols using model 3320

Hydration Status Assessment in Hot-Dry Conditions

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Urine samples were collected following BASE, DEH, REH, and TT in the hot-dry conditions and at BASE in the thermoneutral conditions to assess baseline hydration status. Urine mass was determined using an electronic balance accurate to the nearest 0.1 g with the mass of the empty container subtracted from the total weighed value. USG was measured at each time point using a refractometer. Urine sodium and potassium concentrations and urine osmolality were measured at each time point in triplicate (Smartlyte, Diamond Diagnostics, Massachusetts, United States) and freezing-point osmometer (Model 3320, Advanced Instruments, Inc., Massachusetts, United States, respectively).
Blood samples were collected in the hot-dry conditions following BASE, DEH, REH, and TT. These samples were collected in serum separator tubes and left in an upright position for 30 min to allow serum clotting to occur. Serum separator tubes were then centrifuged (10 min, 4°C, 4,000 rpm) and serum separated. Serum osmolality was measured immediately after centrifugation at each time point in triplicate using a freezing-point osmometer (Model 3320, Advanced Instruments, Inc., Massachusetts, United States). Hemoglobin and concentrations of hematocrit, glucose, insulin, sodium, potassium, and chloride were analyzed for each time point (Quest Diagnostics) within 2 days of sample collection.

Berry C.W., Wolf S.T., Cottle R.M, & Kenney W.L. (2021). Hydration Is More Important Than Exogenous Carbohydrate Intake During Push-to-the-Finish Cycle Exercise in the Heat. Frontiers in Sports and Active Living, 3, 742710.

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Infant Hormone Risk Assessment Protocol

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Hormone levels are derived from urine samples; the 2-month sample serves as our measure of infant hormonal risk to test Aim 1. Urine osmolality (solute level in urine) will be measured in duplicate using the freezing point depression method (Model 3320, Advanced Instruments, Norwood, MA) to control for infant hydration. Assessment of all infant biomarkers will be done using commercially available ELISAs (leptin; BioVision [Milpitas, CA], adiponectin; Quidel [San Diego, CA], insulin; Sigma [St. Louis, MO]). All samples from an individual will be assayed in a single kit to minimize the effects of inter-assay variability.

Leerkes E.M., Buehler C., Calkins S.D., Shriver L.H, & Wideman L. (2020). Protocol for iGrow (Infant Growth and Development Study): biopsychosocial predictors of childhood obesity risk at 2 years. BMC Public Health, 20, 1912.

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Urinary Protein and Osmolality Analysis

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Urinary proteins level was assessed by Bradford assay (Biorad Protein Assay, Biorad, Segrate, Italy). Urine osmolality was measured by freezing point depression osmometer (Model 3320, Advanced Instruments, inc., MA, United States) as previously reported (Zacchia et al., 2016 (link)). Serum electrolytes were analyzed by Vitrovet (Scil, Treviglio (BG), Italy).

Iervolino A., De La Motte L.R., Petrillo F., Prosperi F., Alvino F.M., Schiano G., Perna A.F., Di Matteo D., De Felice M., Capasso G, & Trepiccione F. (2018). Integrin Beta 1 Is Crucial for Urinary Concentrating Ability and Renal Medulla Architecture in Adult Mice. Frontiers in Physiology, 9, 1273.

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Comprehensive 24-Hour Urine Analysis

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For 24 h hydration status, each 24 h urine sample was assessed for urine volume (U_VOL) to the nearest 0.0001 kg using a digital scale (Ranger 2000, OHAUS Corporation, Parsippany, NY, USA), urine osmolality (U_OSMO) measured in duplicate using the freezing point depression method (Model 3320, Advanced Instruments Inc., Norwood, MA, USA), and urine specific gravity (U_SG) using a digital refractometer (Reichert AR200, Reichert Technologies, Buffalo, NY, USA).

Adams W.M., Wininger M., Zaplatosch M.E., Hevel D.J., Maher J.P, & McGuirt J.T. (2020). Influence of Nutrient Intake on 24 Hour Urinary Hydration Biomarkers Using a Clustering-Based Approach. Nutrients, 12(10), 2933.

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Serum Osmolality Measurement in Mice

Cited 3 times

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As previously described (Mitchell et al., 2018 (link)), blood samples were drawn via cardiac puncture prior to perfusion fixation while mice were deeply anesthetized with isoflurane (3% for 180 seconds). Blood samples (0.3 mL) were centrifuged for 8 minutes at 5000 rpm, and serum was collected to measure osmolality by freezing point depression (Advanced Instruments, Inc., model 3320). Reports are conflicting whether, and over what time course, isoflurane anesthesia might affect inflammatory markers (Wu et al., 2012 (link); Altay et al., 2014 (link)); for the present experiments, all mice were exposed to the same concentration of isoflurane (3% in oxygen) for no longer than 3 minutes immediately prior to perfusion fixation to minimize potential confounds.

Gilman T.L., Mitchell N.C., Daws L.C, & Toney G.M. (2018). Neuroinflammation Contributes to High Salt Intake-Augmented Neuronal Activation and Active Coping Responses to Acute Stress. International Journal of Neuropsychopharmacology, 22(2), 137-142.

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Nanoemulsion Osmolality Measurement

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The osmolality of optimized nanoemulsion was measured by the freezing-point depression method using a semi-micro osmometer (Model 3320, Advanced Instruments, Inc., USA). The instrument was calibrated with reference standards of water for injection and calibration solution NaCl to give values of 0 and 400 mOsm/kg, respectively. The measurements of osmolality were taken three times.

Samiun W.S., Ashari S.E., Salim N, & Ahmad S. (2020). Optimization of Processing Parameters of Nanoemulsion Containing Aripiprazole Using Response Surface Methodology. International Journal of Nanomedicine, 15, 1585-1594.

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Hydration Status Assessment Protocol

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Participants were instructed to provide a pre-exercise nude body mass (BM_nude) measure prior to samples being given. Post-exercise BM_nude was obtained after capillary blood samples were taken and before a urine sample was given. Both BM_nude measures were obtained on every visit and on days one and four of acclimation. Urine samples were collected on days one and four of acclimation and on every visit for HST’s following the same routine as HA measures. Urine specific gravity (SG_u) was calculated using a refractometer (Unicron-N, Urine specific gravity refractometer, Atago CO., Tokyo, Japan) [42 (link)]. Urine colour (colour_u) was measured using a colour_u chart [43 (link)] and urine osmolality (osm_u) was measured using an osmometer (Model 3320, Advanced Instruments Inc., Massachusetts, USA). All measures were collected immediately prior to and post exercise, in duplicate and analysed immediately with the mean value reported.

Shaw J., Walkington C., Cole E., Gleadall-Siddall D.O., Burke R., Bray J., Simpson A.J., Vince R.V, & Garrett A.T. (2022). Effectiveness of short-term isothermic-heat acclimation (4 days) on physical performance in moderately trained males. PLOS ONE, 17(11), e0270093.

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Measuring Urinary Osmolality by Freezing Point

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Urinary osmolality was measured by freezing point depression osmometer (Model 3320, Advanced Instruments, Inc., MA, United States) as previously^{52 (link)}.

Prosperi F., Suzumoto Y., Marzuillo P., Costanzo V., Jelen S., Iervolino A., Guarino S., La Manna A., Miraglia Del Giudice E., Perna A.F., Zacchia M., Cordat E., Capasso G, & Trepiccione F. (2020). Characterization of five novel vasopressin V2 receptor mutants causing nephrogenic diabetes insipidus reveals a role of tolvaptan for M272R-V2R mutation. Scientific Reports, 10, 16383.

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Plasma Biomarker Measurements in Aquatic Organisms

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Plasma concentration of cortisol was determined by radioimmunoassay as described in (Sundh et al., 2011 (link)) (modified from Young, 1986 (link)). Plasma osmolality was determined using a micro‐osmometer (Model 3320; Advanced Instruments). Total plasma potassium (K⁺), sodium (Na⁺) and calcium (Ca²⁺) were measured from whole plasma using a flame emission photometer, with LiCl as internal standard (Eppendorf AG, Hamburg, Germany; model ELEX 6361) (Sillanpää et al., 2016 (link)). Haemolymph glucose was measured with commercially available enzymatic test kit (Glucose HK; Sigma‐Aldrich), with protocols adapted to a 96‐well microplate (Schram et al., 2010 (link)). Samples were first diluted in distilled water (1:6), and 10 μl of the diluted sample or standard (5.55 mmol/L glucose) was mixed with 200 μl reagent provided with the kit and incubated for 15 min at 20°C. Absorbance was read within 30 min at 340 nm on a SpectraMax 190 Microplate Reader (Molecular Devices).

Green L., Faust E., Hinchcliffe J., Brijs J., Holmes A., Englund Örn F., Svensson O., Roques J.A., Leder E.H., Sandblom E, & Kvarnemo C. (2022). Invader at the edge — Genomic origins and physiological differences of round gobies across a steep urban salinity gradient. Evolutionary Applications, 16(2), 321-337.

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Measurement of Urinary Parameters in Water-Deprived Mice

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Mice were acclimated for few days in metabolic cages (3600M021; Techniplast, West Chester, PA, USA). Following acclimation and baseline measurements of urinary parameters, mice were water deprived for 24 hours, as we did previously [25 (link)]. Tails were clipped with sterilized scissors to collect 50 μl blood into heparinized micro-hematocrit capillary tubes (Fischer, Cat. # 22-362-566) before and after water deprivation. Urine and plasma osmolarity was measured in duplicates using freezing point depression osmometer Model 3320 (Advanced Instruments, Norwood, MA, USA). Urinary creatinine concentration was assessed with QuantiChrom Creatinine Assay Kit (DICT-500; BioAssay Systems, Hayward, CA, USA) utilizing the improved Jaffe method, as we did before [30 (link)]. Urinary AVP levels were assayed with Vasopressin Arg⁸-Vasopressin ELISA kit (Enzo Life Sciences, Farmingdale, NY, USA) following the manufacturer’s protocols. Urinary Ca²⁺ concentration was measured using a Jenway PFP7 Flame photometer (Bibby Scientific, Burlington, NJ, USA).

Tomilin V.N., Mamenko M., Zaika O., Ren G., Marrelli S.P., Birnbaumer L, & Pochynyuk O. (2019). TRPC3 determines osmosensitive [Ca2+]i signaling in the collecting duct and contributes to urinary concentration. PLoS ONE, 14(12), e0226381.

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