![]() One of the widely employed strategies is to slice the tissues and analyze layer-by-layer. It remains challenging to synchronously unite accuracy and large-scale acquirement of information through optical scanning with current technologies. ![]() Overall, this method improves immunofluorescence signal sensitivity, specificity and stability in the whole brain of juvenile and adult zebrafish, which is applicable for fine structural analysis, neural circuit mapping and three-dimensional cell counting.ĭue to the complexity of the biological systems such as the brain, it is essential to image the whole organisms or organs at cellular or molecular levels to study their structures and functions. Moreover, both the structure and the fluorescent signal persist for at least 16 months with excellent signal retention ratio. Compared to traditional method, TSA-PACT achieves approximately tenfold signal amplification and twofold improvement in signal-to-noise ratio (SNR). We show that TSA-PACT is able to reduce the opacity of the zebrafish brain by more than 90% with well-preserved structure. To address this issue, we developed TSA-PACT, a method combining tyramide signal amplification (TSA) and PACT, to transform samples into hydrogel polymerization frameworks with covalent fluorescent biomarkers assembled. However, it has been difficult to acquire high quality results through immunofluorescence (IF) staining on the cleared samples. CLARITY, PACT) provide new solutions by homogenizing the refractive index of the samples to create transparency. ![]() Recent advances in tissue clearing technology (e.g. It requires large-scale volumetric imaging in cellular or molecular resolution, which could be quite challenging. The Creative Commons Public Domain Dedication waiver ( ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.įor comprehensive studies of the brain structure and function, fluorescence imaging of the whole brain is essential. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made.
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