An article An advanced optical clearing protocol allows label-free detection of tissue necrosis via multiphoton microscopy in injured whole muscle WOS:000604981700024 published article about 2ND-HARMONIC GENERATION; SHG MICROSCOPY; SKELETAL-MUSCLE; HIGH-RESOLUTION; MICROARCHITECTURE; REGENERATION; MORPHOMETRY; EXCITATION in [Schneidereit, Dominik; Broellochs, Anita; Ritter, Paul; Kreiss, Lucas; Schuermann, Sebastian; Friedrich, Oliver] Friedrich Alexander Univ Erlangen Nurnberg, Inst Med Biotechnol, Paul Gordan Str 3, D-91052 Erlangen, Germany; [Schneidereit, Dominik; Ritter, Paul; Kreiss, Lucas; Schuermann, Sebastian; Friedrich, Oliver] Friedrich Alexander Univ Erlangen Nurnberg, Grad Sch Adv Opt Technol SAOT, Paul Gordan Str 7, D-91052 Erlangen, Germany; [Mokhtari, Zeinab; Beilhack, Andreas] Wurzburg Univ, Med Sch, Interdisciplinary Ctr Clin Res, Lab Expt Stem Cell Transplantat,Dept Med 2, Zinklesweg 10, D-97078 Wurzburg, Germany; [Kroenke, Gerhard; Ackermann, Jochen A.; Faas, Maria; Grueneboom, Anika] Friedrich Alexander Univ Erlangen Nurnberg, Clin Internal Med 3, Ulmenweg 18, D-91054 Erlangen, Germany; [Kroenke, Gerhard; Ackermann, Jochen A.; Faas, Maria; Grueneboom, Anika] Friedrich Alexander Univ Erlangen Nurnberg, Clin Rheumatol & Immunol, Ulmenweg 18, D-91054 Erlangen, Germany; [Schuermann, Sebastian; Friedrich, Oliver] Muscle Res Ctr Erlangen MURCE, Henkestr 91, D-91052 Erlangen, Germany; [Friedrich, Oliver] Univ New South UNSW, Sch Med Sci SOMS, Kensington Campus,High St, Sydney, NSW 2052, Australia in 2021, Cited 60. The Name is Benzyl ether. Through research, I have a further understanding and discovery of 103-50-4. Product Details of 103-50-4
Rationale: Structural remodeling or damage as a result of disease or injury is often not evenly distributed throughout a tissue but strongly depends on localization and extent of damaging stimuli. Skeletal muscle as a mechanically active organ can express signs of local or even systemic myopathic damage, necrosis, or repair. Conventionally, muscle biopsies (patients) or whole muscles (animal models) are mechanically sliced and stained to assess structural alterations histologically. Three-dimensional tissue information can be obtained by applying deep imaging modalities, e.g. multiphoton or light-sheet microscopy. Chemical clearing approaches reduce scattering, e.g. through matching refractive tissue indices, to overcome optical penetration depth limits in thick tissues. Methods: Here, we optimized a range of different clearing protocols. We find aqueous solution-based protocols employing (20-80%) 2,2′-thiodiethanol (TDE) to be advantageous over organic solvents (dibenzyl ether, cinnamate) regarding the preservation of muscle morphology, ease-of-use, hazard level, and costs. Results: Applying TDE clearing to a mouse model of local cardiotoxin (CTX)-induced muscle necrosis, a complete loss of myosin-II signals was observed in necrotic areas with little change in fibrous collagen or autofluorescence (AF) signals. The 3D aspect of myofiber integrity could be assessed, and muscle necrosis in whole muscle was quantified locally via the ratios of detected AF, forward- and backward-scattered Second Harmonic Generation (fSHG, bSHG) signals. Conclusion: TDE optical clearing is a versatile tool to study muscle architecture in conjunction with label-free multiphoton imaging in 3D in injury/myopathy models and might also be useful in studying larger biofabricated constructs in regenerative medicine.
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