Fluorescent Silver Staining of Proteins in Polyacrylamide Gels

Wong, Alex Y. H.; Xie, Sheng; Tang, Ben Zhong; Chen, Sijie

doi:10.3791/58669

Cited by 3 publications

(2 citation statements)

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“…in this study for the reason that it uses the same monomers of acrylamide and bis-acrylamide as the polyacrylamide gel which are compatible with TPE-4TA stain. 28,29 After tissue clearing with passive CLARITY, silver-AIE staining was carried out with double silver impregnation.…”

Section: Workflow Of Fluorogenic Silver-aie Staining In Hydrogel-or Paraffin-embedded Brain Sectionsmentioning

confidence: 99%

“…We have recently proposed a fluorogenic silver-AIE staining method and utilized AIEgen TPE-4TA to visualize proteins in the silver-impregnated polyacrylamide gel. 28,29 Taking the advantages of Ag + binding by tetrazole-functionalized TPE-4TA accompanied with fluorescence emission by the AIE properties, here we explore the strategy of fluorogenic AIE-based silver staining in brain slices in addressing the lack of an efficient and reliable staining method in large-scale brain analysis. A staining protocol modified from the classic silver staining methods (especially the Bielschowsky and Cajal's stain) is established and silver-impregnated brain tissues are analyzed by using advanced fluorescence microscopy.…”

Section: Introductionmentioning

confidence: 99%

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Fluorescent Silver-AIE Visualization of Neurons and Fibres in the Cleared Brain

Xie

Wong

Kam

et al. 2021

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Brain-mapping projects for reconstructing structures and networks in three-dimensions (3D) demand costeffective, robust, and histologically homogeneous chemical stains for the applications of high-throughput staining in large tissues. Current staining methods such as immunohistology are practically unsustainable for large-scale specimens due to their limited dye-permeability which leads to inhomogeneous performance in thick samples. Herein, we report a novel fluorogenic visualization method for mouse brain tissues by making use of a Ag + -specific aggregation-induced emission (AIE) strategy after silver impregnation. This novel fluorogenic silver-AIE method surpasses the chromogenic detection used by many conventional silver staining protocols, which have historically made revolutionary contributions to neuroanatomy. This fluorescent silver-based stain allows for analysis of images acquired using wide-field, confocal, SIMs, or light-sheet fluorescence microscopy. We validated the sensitivity, specificity, and versatility of the silver-AIE stain as compared to conventional silver stains and other neuron-specific ones. The fluorogenic AIE method eliminates the high background, artefacts, and poor reproducibility often associated with the reduction of silver solutions. The silver-AIE stain is demonstrated to be highly compatible with immunofluorescence co-staining in paraffin sections and can be applied to hydrogel-based cleared tissue by the passive CLARITY method for image analysis of large tissue slices of 300 μm thick in this study.From this fluorogenic strategy, it is possible for vast types of classic silver staining methods to be modified and re-vitalized.

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Section: Workflow Of Fluorogenic Silver-aie Staining In Hydrogel-or Paraffin-embedded Brain Sectionsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%