Urine was collected overnight into specimen vials (animals were placed in metabolism cages at the end of the working day preceding the day of urine collection). The following parameters were determined using a Cobas u 411 semi-automated test strip analyser (Roche): specific gravity (SG1), protein (Prot), ketones (Keto), volume (Vol), creatinine (U-Creat), bilirubin (Bili), colour (Col), glucose (U-Gluc), erythrocytes (U-RBC), appearance (App), nitrite (Nite), pH, leukocytes (U-WBC).
Cobas u 411
Manufactured by Roche
Sourced in Germany
The Cobas u 411 is an automated urine analyzer developed by Roche. It is designed to perform routine urinalysis testing, including the measurement of various urine parameters such as pH, specific gravity, and the detection of certain analytes. The Cobas u 411 is intended to provide reliable and efficient urine analysis for clinical laboratories.
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8 protocols using cobas u 411
1
Comprehensive Urine Analysis Protocol
2
Urinalysis of Animals During Treatment
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Urinalysis was performed on all animals in the main groups during the 13th week of treatment and in the recovery groups during the 4th week of recovery. Freshly voided urine (urine voided within 3 h) was collected, and the following indicators were measured using a Combur10Test®M stick (Roche, Germany) and a urine chemistry analyzer (cobas u 411, Roche, Germany): pH, protein, glucose, ketone body, bilirubin, and occult blood. Urine color and transparency were examined macroscopically, and urine sediment was examined microscopically. Concentrated urine (urine voided within approximately 24 h) was collected to measure volume and specific gravity using a measuring cylinder and a refractometer (Vet360, Reichert, USA), respectively.
3
Automated Urine Analysis: Comparing Methods
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The cobas u 411 and Urisys 2400 analyzers (Roche Diagnostics GmbH, Mannheim, Germany) are urine test strip analyzers that use reflectance photometry. Urisys 2400 test strip cassettes were used with the Urisys 2400 analyzer and Combur [10 (link)] Test® M test strips were used with the cobas u 411 analyzer.
Visual microscopy using KOVA® chamber technology (Cat. No. 3518345001, Kova International Inc., Garden Grove, CA), a standardized methodology according to European urinalysis guidelines [14 (link)], was used as a harmonized comparison method across the three evaluation centers. Two KOVA slides of each sample were prepared and counted using bright field microscopy, each by two experienced operators, to reduce counting error; the mean value was then calculated. Further details on the KOVA procedure are provided inSupplementary Material S2 .
The iQ200 analyzer (Iris Diagnostics, Chatsworth, CA) is a second-generation automated microscopy analyzer that captures images from planar flow of urine particles. A neural network (Auto-Particle Recognition™) is used to classify and quantify particles in the sample; 500 photographs are taken from each urine sample and compared with standard images.
Visual microscopy using KOVA® chamber technology (Cat. No. 3518345001, Kova International Inc., Garden Grove, CA), a standardized methodology according to European urinalysis guidelines [14 (link)], was used as a harmonized comparison method across the three evaluation centers. Two KOVA slides of each sample were prepared and counted using bright field microscopy, each by two experienced operators, to reduce counting error; the mean value was then calculated. Further details on the KOVA procedure are provided in
The iQ200 analyzer (Iris Diagnostics, Chatsworth, CA) is a second-generation automated microscopy analyzer that captures images from planar flow of urine particles. A neural network (Auto-Particle Recognition™) is used to classify and quantify particles in the sample; 500 photographs are taken from each urine sample and compared with standard images.
4
Comprehensive Metabolic Profiling in Rodents
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Animals in the cage were separated and transferred to a metabolic cage or a cage equipped with a device for collection of urine on the day before administration as well as terminal sacrifice (day 29), and recovery sacrifice (day 43). Before urine collection, animals were fasted overnight; however, drinking water was made available. The urine volume was recorded using a measuring cylinder, and the following parameters were measured using a Cobas U411 urine analyzer (Roche, Switzerland) using a urine reagent strip: color, clarity, pH, specific gravity, bilirubin, proteins, urobilinogen level, nitrite level, glucose level, erythrocyte count, ketone, leukocyte count, urine potassium, urine chloride, urine sodium, urine cast and epithelial cell count.
5
Hormonal Profile Assessment in Patients
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All patients had routine blood draws to measure serum prostate-specific antigen (PSA), C-reactive protein (CRP), estrogen, testosterone (normal range: 300–1000 ng/dL), and estradiol. In the central laboratory of the Giessen University Hospital (ADVIA and ADVIA Centaur, Siemens Health Care), routine laboratory procedures were used to evaluate the levels of prolactin, sex hormone-binding globulin (SHBG), albumin, follicle-stimulating hormone (FSH), lung tanning hormone (LH), and albumin in parallel if a decreased testosterone level was discovered. Leukocyturia was identified using an automated quantitative urine particle analyzer (cobas u 411, Roche Diagnostics GmbH) and a urine dipstick. A technician who was blind to the sources of the samples conducted the assays.
6
Urine Analysis for Metabolic Parameters
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Urine was collected in specimen vials using a metabolism cage. The following parameters were determined using a Cobas u 411 semi-automated test strip analyser (Roche): specific gravity (SG1), volume (Vol), colour (Col), appearance (App), pH, nitrite (Nite), protein (Prot), glucose (U-Gluc), ketones (Keto), urobilinogen (Urob), bilirubin (Bili), erythrocytes (U-RBC), leukocytes (U-WBC).
7
Comprehensive Metabolic Profiling in Diabetic Ketoacidosis
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All anthropometric and laboratory measures were obtained upon admission. The anthropometric information included age, gender, BMI, SBP, DBP, HR, and history of T2DM. Laboratory results included serum ketones (β-hydroxybutyrate), urine ketones (acetoacetic acid and acetone), Hb, HbA1c, PG, insulin, C-peptide, liver function tests (ALT and AST), renal function tests (sCr and BUN), UA, CRP, lipid profile (TG, TC, LDL, HDL, and FFA), arterial pH, base excess (BE), bicarbonate, electrolyte levels, and osmolality (Table 1 ). Serum ketones were measured using a MediSense hand-held device (Abbott Corporation, Abbott Park, IL, USA). Urine ketones were measured using the nitroprusside method (Semi-automatic urine analyzer, Cobas u411, Roche, Germany). HbA1c was measured by high performance liquid chromatography (HLC-723G8, Tosoh, Japan). OGTTs were conducted prior to any treatment in all but 40 patients with diabetic ketoacidosis (after correction of acidosis). Insulin and C-peptide were measured during the OGTT. The formula, [sodium (mEq/l) × 2 + glucose (mg/dl)/18]34 (link)49 (link), was used to calculate effective osmolality.
8
Comprehensive Urinalysis: Physical, Chemical, and Microscopic Examination
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Physical, chemical and microscopic examination of urine samples were performed by experienced laboratory personnel. Urine samples were delivered in the laboratory in urine cups or tubes (Greiner Bio-One GmbH, Kremsmu¨nster, Austria). Combur-10 Test Õ strips (Roche Diagnostics, GmbH, Mannheim, Germany) were used for chemical examination of urine. Urine analyzers Cobas u411 and Miditron Junior II (both Roche Diagnostics, GmbH, Mannheim, Germany) were used as semi-automated urinalysis system for test strips scanning. Tests were performed as described in manufacturer's instruction.
For sediment preparation, 10 mL of urine was centrifuged at 500 Â g for 10 min on Hettich Rotofix 32 A benchtop centrifuge (Hettich Lab Technology, Beverly, United States). After discarding supernatant, the sediment was resuspended in 500 L of urine, dropped on glass slide and covered by coverslip. All microscopic examinations were performed by the bright-field microscopy on native urine sediment. Firstly, examination was done under lower magnification of 100 Â and fields were scanned for casts. Afterwards, 400 Â magnification was used for examination of other elements in urine on 10 random fields.
For sediment preparation, 10 mL of urine was centrifuged at 500 Â g for 10 min on Hettich Rotofix 32 A benchtop centrifuge (Hettich Lab Technology, Beverly, United States). After discarding supernatant, the sediment was resuspended in 500 L of urine, dropped on glass slide and covered by coverslip. All microscopic examinations were performed by the bright-field microscopy on native urine sediment. Firstly, examination was done under lower magnification of 100 Â and fields were scanned for casts. Afterwards, 400 Â magnification was used for examination of other elements in urine on 10 random fields.
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