Basal tear samples were carefully obtained to avoid touching the ocular surfaces from the inferior tear meniscus of both eyes by using glass capillary tubes (Corning, Inc., Corning, NY, USA) or micropipettes (Eppendorf, Hamburg, Germany) before and immediately after 1 h and 4 h of smartphone use as described previously.[20 (link)] Twenty microliter tear samples were obtained and diluted with phosphate-buffered saline. Tear samples were placed in microtubes and stored at −70°C until further examination.
Glass capillary tubes
Manufactured by Corning
Sourced in United States
Glass capillary tubes are small, hollow glass cylinders used in various laboratory applications. They are designed to facilitate the precise handling and transfer of small volumes of liquids or gases. The tubes have a uniform and precise internal diameter, allowing for controlled fluid flow and measurement.
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Lab products found in correlation
3 protocols using glass capillary tubes
1
Basal Tear Collection and Analysis
2
Quantifying Tear Neuromediators Using ELISA
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Tear samples were carefully obtained from the inferior tear meniscus of the right eye using glass capillary tubes (Corning, Inc., Corning, NY, USA) to avoid touching the ocular surfaces after examination of their clinical parameters. Two hundred microliters of tear samples were obtained and diluted with phosphate-buffered saline (1:4). The samples were placed in microtubes and stored at −70℃ until further examination.18 (link)
A specific enzyme-linked immunosorbent assay was performed on extracted proteins to quantify the concentrations of neuromediators in the tear samples of subjects following manufacturer instructions. Tear samples were centrifuged at 3000 g for 10 min and filtered through a 0.45-mm filter before the enzyme-linked immunosorbent assay. Protein samples (100 µL, 10 µg/mL) of NGF (Elabscience, Houston, TX, USA), serotonin (Abcam, Cambridge, UK), CGRP (Elabscience), substance P (R&D SYSTEMS, Minneapolis, MN, USA), NPY (Elabscience), and VIP (Elabscience) were absorbed onto each conjugate-coated plate. Levels of neuromediators in a given sample were measured by comparing its absorption with the standard curve. The minimum detectable concentrations of NGF, serotonin, CGRP, substance P, NPY, and VIP were 15.63 pg/mL, 0.5 ng/mL, 15.63 pg/mL, 78.1 pg/mL, 31.25 pg/mL, and 7.81 pg/mL, respectively.
A specific enzyme-linked immunosorbent assay was performed on extracted proteins to quantify the concentrations of neuromediators in the tear samples of subjects following manufacturer instructions. Tear samples were centrifuged at 3000 g for 10 min and filtered through a 0.45-mm filter before the enzyme-linked immunosorbent assay. Protein samples (100 µL, 10 µg/mL) of NGF (Elabscience, Houston, TX, USA), serotonin (Abcam, Cambridge, UK), CGRP (Elabscience), substance P (R&D SYSTEMS, Minneapolis, MN, USA), NPY (Elabscience), and VIP (Elabscience) were absorbed onto each conjugate-coated plate. Levels of neuromediators in a given sample were measured by comparing its absorption with the standard curve. The minimum detectable concentrations of NGF, serotonin, CGRP, substance P, NPY, and VIP were 15.63 pg/mL, 0.5 ng/mL, 15.63 pg/mL, 78.1 pg/mL, 31.25 pg/mL, and 7.81 pg/mL, respectively.
3
Oxidative Stress Biomarkers in Tear Fluids
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As we previously described, basal tear secretions were harvested atraumatically from the both inferior tear meniscus using glass capillary tubes (Corning, Inc., Corning, NY) [23 (link)]. Caution was taken not to irritate the surfaces of the conjunctival and corneal. Fifty-microliter of tear materials were harvested and diluted by phosphate-buffered saline. Tear fluids were stored in microtubes and kept at − 70 °C until additional examination.
Total protein levels of the 8-OHdG (Cell Biolabs, San Diego, CA, USA) and MDA (Cell Biolabs) were detected using enzyme-linked immunosorbent assays (ELISA) in accordance with the producer’s instructions [24 (link)]. Tear specimens were centrifuged at 3000 g during 10 min or filtered through 0.45 mm filter, prior to use in the ELISA. Both 8-OHdG and MDA protein samples (50 μl) were absorbed onto the 8-OHdG and MDA conjugate coated plate, respectively. The level of 8-OHdG and MDA in an unknown sample was measured by comparing its absorption with the known standard curve. The minimal detectable concentrations of the 8-OHdG and MDA were above 0.078 ng/mL and 6 pmol/mg, respectively.
Total protein levels of the 8-OHdG (Cell Biolabs, San Diego, CA, USA) and MDA (Cell Biolabs) were detected using enzyme-linked immunosorbent assays (ELISA) in accordance with the producer’s instructions [24 (link)]. Tear specimens were centrifuged at 3000 g during 10 min or filtered through 0.45 mm filter, prior to use in the ELISA. Both 8-OHdG and MDA protein samples (50 μl) were absorbed onto the 8-OHdG and MDA conjugate coated plate, respectively. The level of 8-OHdG and MDA in an unknown sample was measured by comparing its absorption with the known standard curve. The minimal detectable concentrations of the 8-OHdG and MDA were above 0.078 ng/mL and 6 pmol/mg, respectively.
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