Ten cassava leaves (counting from the top) were harvested at the end of the day or end of the night from each plant, cleared in 80% (v/v) ethanol, rinsed in water and iodine-stained with Lugol’s solution (Sigma-Aldrich).
Lugol s solution
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom
Lugol's solution is an aqueous mixture of potassium iodide and elemental iodine. It is a laboratory reagent used for various applications, such as staining and detection procedures.
Automatically generated - may contain errors
Lab products found in correlation
Improved neubauer, by Boeco (3 mentions)
Crystal violet, by Merck Group (2 mentions)
Sodium hydroxide (naoh), by Merck Group (2 mentions)
Vivact 40, by Scanco (1 mentions)
Formical 2000, by StatLab (1 mentions)
Skyscan 1176, by Bruker (1 mentions)
Formalin solution, by Merck Group (1 mentions)
Nrecon software, by Bruker (1 mentions)
Mayer s hematoxylin, by Merck Group (1 mentions)
Bacto tryptic soy broth, by BD (1 mentions)
Sodium nitrite, by Merck Group (1 mentions)
Masson s trichrome stain kit, by Abcam (1 mentions)
2 2 2 tribromoethanol, by Merck Group (1 mentions)
Tert amyl alcohol, by Merck Group (1 mentions)
Live dead baclight bacterial viability kit, by Thermo Fisher Scientific (1 mentions)
Sodium alginate, by Merck Group (1 mentions)
Glutathione reduced form, by Fujifilm (1 mentions)
Amersham imagequant, by Cytiva (1 mentions)
Eclipse 90i, by Nikon (1 mentions)
Glass fiber filter, by Cytiva (1 mentions)
35 protocols using lugol s solution
1
Cassava Leaf Starch Visualization
2
Visualizing Subchondral Bone Resorption
3
Micro-CT Imaging of Mouse Embryos
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WT or Clspn+/− pairings were mated overnight. Then, male mice were removed (E0.5) and the females were checked for a mucus vaginal plug. At E13.5, female mice were humanely sacrificed by cervical dislocation and the embryos harvested. The yolk sac was removed for genotyping and the embryos fixed in 10% Formalin solution (HT501850, Sigma) for 24 h. Following this, the embryos were embedded in 1% (w/v) agarose and stained with Lugol's solution (1.09261, Sigma) for a further 24 h. Following this, micro-CT was performed on the embryos in collaboration with the Newcastle Preclinical In Vivo Imaging Facility using the Skyscan 1176, Bruker-microCT system. All samples were acquired at an isotropic resolution of 9 μm using the following parameters: a source voltage of 45 kV, source current of 550 μA, 1315 ms exposure time, 0.5 m Aluminium filter, 0.3° rotation step using 360 projections, 4 frame averaging. The total scan duration was 3 h. Image reconstruction was performed using NRecon software (Bruker).
4
Comprehensive Reagent Procurement for Multidisciplinary Research
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Glutathione (reduced form) was purchased from Wako Pure Chemical (Osaka, Japan). Pectin and agar were purchased from Yakuri Co., Ltd. (Osaka, Japan). Sodium alginate, PEG (average molecular weight: 8000), sodium nitrite, crystal violet, Lugol’s solution, 2,2,2 tribromoethanol, tert-amyl alcohol, Mayer’s hematoxylin, and eosin-Y disodium were purchased from Sigma-Aldrich (St. Louis, MO, USA). Bacto™ tryptic soy broth (TSB) and Difco™ cetrimide-agar media were purchased from BD Biosciences (San Jose, CA, USA). A Twort’s Gram stain set was purchased from Newcomer Supply (Middleton, WI, USA). Masson’s trichrome stain kit was purchased from Abcam (Cambridge, MA, USA). The LIVE/DEAD®BacLight™ bacterial viability kit was purchased from Thermo Fisher Scientific (Waltham, MA, USA). All other reagents and solvents were of the highest analytical grade commercially available.
5
Starch-Degrading Activity in Dairy Products
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Starch-degrading activity in dairy products or of purified enzymes was determined with starch zymography. Chocolate pudding aroma as well as liquefied and control chocolate pudding were diluted 1:2 with 35 mM NaCl for homogenization. An amount of 20 µg protein per pudding preparation as well as 0.005–20 µL of the aroma preparation were subjected to 1D SDS-PAGE under denaturing, but non-reducing conditions on 12% gels at 4 °C. SDS gels were incubated overnight in a 0.2 M phosphate buffer (pH 6) containing 1% starch at 4 °C, followed by 2 h incubation at 45 °C with fresh phosphate-starch buffer. Gels were washed in Aqua bidest and subsequently incubated in Lugol’s solution (Sigma-Aldrich) at room temperature until light bands became visible. The stained gels were washed again in Aqua bidest. Starch-degrading activity was detected as transparent bands on a dark background of undegraded substrate. Gels were scanned with an Amersham ImageQuant 800 biomolecular imager (GE Healthcare).
6
Algal Cell Density and Viability Assay
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For cell density estimation, 10 μL of algal culture was mixed with 30 μL of Lugol’s solution (Sigma-Aldrich, St. Louis, MO, United States) and the cell number was counted in duplicate using a light microscope (BX43, Olympus, Tokyo, Japan) and a hemacytometer (Improved Neubauer, Boeco, Germany) as mentioned above. The cell density was calculated according to the manufacturer’s manual and expressed as units of 106⋅ mL–1.
After 6 h of treatments, the viability of algal cells was estimated by loading 2 μL of cell suspension on TAP agar plates, and incubating the plates for 72 h at 28°C under illumination at 50 μmol⋅ m–2⋅ s–1 intensity. The colonies were imaged with a digital Nikon camera, and the final composite images were constructed using Adobe Photoshop (Adobe Systems, San Jose, CA, United States). Together with cell density curve, the cell viability assessed by growth ability (the color and size of the colony) was used to evaluate the effects of chemical challenges.
After 6 h of treatments, the viability of algal cells was estimated by loading 2 μL of cell suspension on TAP agar plates, and incubating the plates for 72 h at 28°C under illumination at 50 μmol
7
Quantifying Trichodesmium Colonies and Pigments
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Individual Trichodesmium colonies for cell counts (using microscopes Eclipse 90i, Nikon Instruments, Düsseldorf, Germany/Axiovert 135, Zeiss, Jena, Germany) were preserved in Lugol's solution (Sigma Aldrich). To quantify the number of cells per colony, the cumulative length of all trichomes in a colony was divided by the average cell length. Samples for analysis of species composition were coated with 0.1% agarose and stained with 4,6-diamidino-2-phenylindole before epi-fluorescence microscopy (Axioplan2 imaging, Zeiss). Trichodesmium species were identified based on cell shape and size following Hynes et al. (2012) (link). For quantification of chl a concentrations, 15 colonies per replicate were collected on glass fiber filters (25 mm diameter, Whatman, Maidstone, UK), and the chl a extracted in 5 ml of 90% acetone at −20 °C before fluorometric analysis (Turner Designs 10-AU, Sunnyvale, CA, USA; Strickland and Parsons, 1972 ). For determination of particulate organic C and N (POC and PON) contents, as well as dry weight, 15–25 colonies per replicate were collected on pre-combusted glass fiber filters (25 mm, Whatman). Before mass spectrometry (ThermoFinnigan DeltaXP, Bremen, Germany), filters were acidified (HCl fume, >12 h) and dehydrated (50 °C, >12 h).
8
Starch Staining of Tobacco Leaves
9
Histochemical Assays for Plant Seedling Analysis
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For β-glucuronidase (GUS) staining, 5-day-old etiolated seedlings were vacuum-infiltrated with a solution consisting of 50 mM sodium phosphate buffer pH 7.2, 250 µM K3Fe(CN)6, 250 µM K4Fe(CN)6, 2% Triton X-100, and 1 mM 5-bromo-4-chloro-3-indolyl-β-D-glucuronic acid (X-GlcA, Duchefa) for 1 h and then incubated at 37 °C until color development.
For starch detection, 5-day-old etiolated seedlings were incubated for 5 min in Lugol’s solution (Sigma-Aldrich), followed by 10 min washing with water.
Improved propidium iodide staining was done as described in (Truernit et al., 2008 (link)) with modifications (80 °C prewarmed ethanol was applied for 1 min and Hoyer’s solution was not used).
Cuticular defects were detected using an aqueous solution of 0.05% toluidine blue (Sigma-Aldrich) for 10 min, followed by washing with water.
For starch detection, 5-day-old etiolated seedlings were incubated for 5 min in Lugol’s solution (Sigma-Aldrich), followed by 10 min washing with water.
Improved propidium iodide staining was done as described in (Truernit et al., 2008 (link)) with modifications (80 °C prewarmed ethanol was applied for 1 min and Hoyer’s solution was not used).
Cuticular defects were detected using an aqueous solution of 0.05% toluidine blue (Sigma-Aldrich) for 10 min, followed by washing with water.
10
Postnatal Brain Imaging and Lesion Analysis
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Postnatal pups were anesthetized with Avertin (Sigma-Aldrich; T48402) and underwent intracardiac perfusion with PBS and 2% paraformaldehyde (w/v). Brains were immediately placed in 4% PFA/PBS fixative. Brains remained in fixative until staining with Lugol’s solution (Sigma-Aldrich; L6146) for 48 hours and were subjected to μCT imaging. Brains were randomized and scanned by blinded operators using an Xradia Micro-CT system (Xradia MicroXCT-400, Xradia). Images were acquired at 50 kV, 10 W, 721 projections, and 3-second integration per 180° rotation.
For postlesion labeling, the brain image stacks were volume rendered and overlaced with the labeled lesions in the Avizo 3D environment. All tissue processing, imaging, and volume quantification were done in a blinded manner by investigators at Tianjin Medical University without any knowledge of experimental details.
For postlesion labeling, the brain image stacks were volume rendered and overlaced with the labeled lesions in the Avizo 3D environment. All tissue processing, imaging, and volume quantification were done in a blinded manner by investigators at Tianjin Medical University without any knowledge of experimental details.
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