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урок по технологии 6 класс фгос интерьер жилого дома

урок по технологии 6 класс фгос интерьер жилого дома

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урок по технологии 6 класс фгос интерьер жилого дома, Advances in Thin Tissue Golgi-Cox Impregnation: Fast, Reliable Methods , Step by Step Golgi-Cox Staining for Cryosection - Frontiers, Golgi-Cox Staining Step by Step - PMC, Golgi-Cox Staining Protocol for Neurons and Processes, A modified and highly sensitive Golgi–Cox method to enable complete and , Golgi Stain - Histology, A modified Golgi staining protocol for use in the human brain stem and .
Sections selected for TH and D1R dual-label immunoEM were rinsed several times in 0.1M PB then incubated in a 1% solution of sodium borohydride for 30 minutes at RT. After rinsing, sections were blocked in 10% NGS (Invitrogen, Inc.) containing 0.03% Tx for 1 hour at RT, then incubated for 36 hours at RT in a solution containing mouse anti-TH (1:1000, Millipore). Following extensive rinsing, sections were incubated in a 10% solution of NGS containing biotinylated goat anti-mouse IgG (1:200, Vector Labs) for 2 hours at RT and, again after rinsing, were incubated with ABC (Vector Labs) for 1 hour at RT. The sections were reacted with 0.05% DAB containing 0.01% H2O2 for 3 minutes. After rinsing, sections were incubated for 36 hours at RT in a solution containing rat anti-D1R (1:500, Millipore). Following extensive rinsing, sections were incubated in a 10% solution of NGS containing biotinylated goat anti-rat IgG (1:200, Vector Labs) for 2 hours at RT and, again after rinsing, were incubated with ABC for 1 hour at RT. The sections were reacted with Vector Slate Grey (SG) (Vector Labs) for 3 minutes. Sections selected for TH and D2R dual-label IHC () were rinsed several times in 0.1M PB then incubated in a 1% solution of sodium borohydride for 30 minutes at RT. After rinsing, sections were blocked in 10% NGS (Invitrogen, Inc.) containing 0.03% Tx for 1 hour at RT, then incubated for 36 hours at RT in a solution containing mouse anti-TH (1:1000, Millipore). Following extensive rinsing, sections were incubated in a 10% solution of NGS containing biotinylated goat anti-mouse IgG (1:200, Vector Labs) for 2 hours at RT and, again after rinsing, were incubated with ABC for 1 hour at RT. The sections were reacted with 0.05% DAB containing 0.01% H2O2 for 3 minutes. After rinsing, sections were incubated for 36 hours at RT in a solution containing rabbit anti-D2R (1:500, Millipore). Following extensive rinsing, sections were incubated in a 10% solution of NGS containing biotinylated goat anti-rabbit IgG (1:200, Vector Labs) for 2 hours at RT and, again after rinsing, were incubated with ABC for 1 hour at RT. The sections were reacted with SG for 3 minutes. After the completion of the IHC reactions, the sections were fixed in a solution of 1% osmium tetroxide (Electron Microscopy Sciences) and 1.5% potassium ferricyanide (Sigma-Aldrich Corp) in 0.1M PB for 30 min at RT in the dark. The sections were dehydrated, stained with 1% uranyl acetate en bloc, and flat embedded in Durcupan epoxy resin (components A–D, Sigma-Aldrich Corp.) between two sheets of aclar fluorohalocarbon film. The epoxy resin was allowed to polymerize at 70°C for 72 hours. An area of known size (0.5 × 0.5 mm) was selected from the striatum. Ultrathin serial sections (silver, ~60–70 nm) were cut (~10 per grid), mounted on formvar-coated copper slot grids, and stained with uranyl acetate and lead citrate prior to viewing. The electron microscopic images were captured on a JEOL 100 CXII electron microscope equipped with a side mounted MegaView digital camera coupled to analySIS® image analysis software., We describe a series of reliable, modified ‘Golgi-Cox’ impregnation methods that complement some existing methods and have several advantages over traditional whole brain ‘Golgi’ impregnation., In the present study, we described a protocol of Golgi-Cox Staining for Cryosection, with the modified cryosection protection solution based on the Golgi-Cox method, which makes cryosection easy to apply in the section of the Golgi-Cox impregnated tissue..
Here, we report in detail all steps of the Golgi-Cox staining method on adult mouse brain vibratome sections required for a reliable high quality staining in an acceptable time frame and with well-preserved tissue quality., Pugh BC and Rossi ML (1993) A paraffin wax technique of Golgi-Cox impregnated CNS that permits the joint application of other histological and immunocytochemical techniques., Here, we provide a modified method using the FD Rapid GolgiStain Kit proceeded by an aldehyde fixation step to achieve stable and successful impregnation of embryonic neurons. 2. Materials and methods. 2.1. Aldehyde fixation treatment., The Golgi stain, also known as the Golgi method or Golgi impregnation, is a histological technique used to visualize neurons in the brain. This staining method was developed by Italian physician Camillo Golgi in 1873., Silver stains are notoriously unpre-dictable and staining of neurons appears random (Pasternak and Woolsey, 1975). In the same section, areas of intense stain can be interspersed with regions of no stain, and a protocol that works well on 1 day may produce negligible results on another., Most researchers today rely on a two-step Golgi-Cox staining procedure (Zhong et al., 2019); chromation (potassium chromate and potassium dichromate solution) followed by silver nitrate impregnation that subsequently allows the formation of silver chromate crystals, visualized as brownish-black structures (Zhang et al., 2010)..
 
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