Direct stochastic optical reconstruction microscopy was carried out as described previously28 (link). Briefly, stochastic photoswitching of immunostained samples was initiated with a buffer containing 50 mmol L−1 2-mercaptoethylamine (M9768, Sigma-Aldrich), 40 µg mL−1 catalase (C3155, Sigma-Aldrich), 500 µg mL−1 glucose oxidase (G7141, Sigma-Aldrich), 50% (w/v) D-glucose (Sigma-Aldrich), in PBS pH 7.4. A 643 nm laser set at 20 mW was used to inactivate the AlexaFluor 647 fluorophore into an off-state prior to stochastic reactivation over the acquisition period. A super-resolved composite of 10,000 images acquired over a period of 300 s at 30 ms exposures was then reconstructed using the ThunderSTORM v1.3 ImageJ plugin, using a linear least square localization method. Coordinates of single emitters were filtered based on localization precision and photon count to discard electronic noise (0 nm < localization precision < 7 nm) and sample noise (localization precision >60 nm).
G7141
Manufactured by Merck Group
Sourced in Germany
The G7141 is a laboratory equipment product manufactured by Merck Group. It is a multi-purpose device designed for various scientific applications. The core function of the G7141 is to provide precise measurement and analysis capabilities for researchers and scientists working in various fields.
Automatically generated - may contain errors
Lab products found in correlation
P8250, by Merck Group (2 mentions)
D glucose, by Merck Group (1 mentions)
2 mercaptoethylamine, by Merck Group (1 mentions)
M9768, by Merck Group (1 mentions)
Synergy h1 hybrid multi mode reader, by Agilent Technologies (1 mentions)
4 aminoantipyrine, by Merck Group (1 mentions)
Amplex red, by Thermo Fisher Scientific (1 mentions)
Chitosan, by Merck Group (1 mentions)
A2153, by Merck Group (1 mentions)
Acetic acid, by Merck Group (1 mentions)
Phenol, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)
Fluorescein isothiocyanate (fitc), by Merck Group (1 mentions)
Amplex red reagent, by Thermo Fisher Scientific (1 mentions)
2 6 dichloroindophenol, by Merck Group (1 mentions)
3 5 dinitrosalicylic acid, by Merck Group (1 mentions)
S3139, by Merck Group (1 mentions)
S9763, by Merck Group (1 mentions)
S0505, by Merck Group (1 mentions)
Coomassie brilliant blue g 250 staining, by Merck Group (1 mentions)
5 protocols using g7141
1
Super-resolution Microscopy of Immunostained Samples
2
Colorimetric Assay for Urinary Glucose
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To assess glucose concentration in the urine, mice were restrained and urine was collected with a pipette and frozen at −20 °C. Samples were treated with glucose oxidase (G7141, Sigma-Aldrich) and peroxidase (P8250, Sigma-Aldrich), and the resulting H2O2 concentration was quantified by a colorimetric assay using glucose as standard and employing 4-aminoantipyrine (A4382, Sigma-Aldrich) and sodium 3,5-dichloro-2-hydroxybenzenesulfonate (D4645, Sigma-Aldrich) as coloring reagents. The plate was read at 520 nm on a Synergy H1 Hybrid Multi-Mode Reader (BioTek, Winooski, VT, USA).
3
Glycogen Quantification via Amplex Red
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Glycogen was measured using Amplex Red method. Briefly, liver samples were homogenized in PBS and incubated with or without 1 mg/mL amyloglucosidase (AMG) in 0.2 M NaOAc, pH 4.8 for 1 h at 37 °C to hydrolyze glycogen into glucose. Samples were incubated with 0.25 U/mL glucose oxidase (G7141, Sigma), 0.17 U/mL horseradish peroxidase (P8250, Sigma), and 20 µM Amplex Red (A36006, Invitrogen) in 1 mM EDTA, 1 mM MgCl2, 10 µM FAD (F6625, Sigma), and 100 mM KH2PO4, pH 6.8 for 10 min in the dark at room temperature. Amplex Red emission (587 nm) was measured following excitation at 535 nm. Free glucose was subtracted from total AMG-hydrolyzable glucose to give glycogen values.
4
Rapid Enzyme-Based Biosensor Fabrication
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A simple and rapid electrode functionalisation process was also developed using a plotter cutter. The mask patterns for the selective activation of PB and GOx on the WE and Ag/AgCl on the RE were created on PET films using the same plotter cutter. The masks were sequentially mounted on the fabricated sensor to selectively expose the regions to be opened (Supplementary Fig. S2 k). Sequential deposition of PB/carbon paste (C2070424P2, SunChemical) on the WE and Ag/AgCl paste (011464, ALS, Japan) on the RE was performed by squeezing the paste over screen masks and curing at 100 °C, each for 5 min. The GOx layer was deposited on the PB layer using the drop-casting method. The GOx solution was prepared by mixing 1% acetic acid (695092, Sigma-Aldrich) and 2% chitosan (C3646, Sigma-Aldrich). GOx (G7141, Sigma-Aldrich; 40 mg per 1 mL of phosphate buffered saline (PBS)) and bovine serum albumin (A2153, Sigma-Aldrich; 10 mg per 1 mL of PBS) were added to the acetic acid–chitosan solution. The GOx solution (20 μL) was dropped on the WE and dried in a vacuum chamber for 20 min for a rapid even evaporation (Supplementary Fig. S2 l).
5
Enzymatic Colorimetric Assays for Biomolecules
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Hydrogen peroxide 30 wt.% (VWR Chemicals), horseradish peroxidase ~150 U/mg (77332, Sigma-Aldrich), absolute ethanol (VWR Chemicals, ≥99.8), microcrystalline cellulose
(BASF Basionics, ≥95%), DMSO (Alfa Aesar, ≥99%), glucose oxidase from Aspergillus niger (G7141, Sigma-Aldrich) (AnGOx), bovine serum albumin (05470, Sigma-Aldrich, ≥96%), Sodium iodide (383112, Sigma-Aldrich, ≥99.5%), sodium phosphate dibasic (S9763, Sigma-Aldrich, ≥99%), sodium phosphate monobasic (S3139, Sigma-Aldrich), sodium carbonate (S7795, Sigma-Aldrich, ≥98%), sodium bicarbonate (S5761, Sigma-Aldrich, ≥99.5%), sodium sulfite (S0505, Sigma-Aldrich, ≥98%), Phenol (328111, Sigma-Aldrich, ≥99%), Amplex ® Red reagent (Invitrogen), 2,6-DichloroindoPhenol (D1878, Sigma-Aldrich), Sephadex G-50 Medium (Sigma-Aldrich), 3,5-Dinitrosalicylic acid (D0550, Sigma-Aldrich, ≥98%), Coomassie Brilliant Blue G-250 staining (Sigma, Deisenhofen, Germany), fluorescein isothiocyanate (F7250, Sigma-Aldrich, ≥90%).
(BASF Basionics, ≥95%), DMSO (Alfa Aesar, ≥99%), glucose oxidase from Aspergillus niger (G7141, Sigma-Aldrich) (AnGOx), bovine serum albumin (05470, Sigma-Aldrich, ≥96%), Sodium iodide (383112, Sigma-Aldrich, ≥99.5%), sodium phosphate dibasic (S9763, Sigma-Aldrich, ≥99%), sodium phosphate monobasic (S3139, Sigma-Aldrich), sodium carbonate (S7795, Sigma-Aldrich, ≥98%), sodium bicarbonate (S5761, Sigma-Aldrich, ≥99.5%), sodium sulfite (S0505, Sigma-Aldrich, ≥98%), Phenol (328111, Sigma-Aldrich, ≥99%), Amplex ® Red reagent (Invitrogen), 2,6-DichloroindoPhenol (D1878, Sigma-Aldrich), Sephadex G-50 Medium (Sigma-Aldrich), 3,5-Dinitrosalicylic acid (D0550, Sigma-Aldrich, ≥98%), Coomassie Brilliant Blue G-250 staining (Sigma, Deisenhofen, Germany), fluorescein isothiocyanate (F7250, Sigma-Aldrich, ≥90%).
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