ArticleCell & bioscience2023
TSA-PACT: a method for tissue clearing and immunofluorescence staining on zebrafish brain with improved sensitivity, specificity and stability.
Article in Cell & bioscience, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed, 7 citations in OpenAlex.
- Article
- EpiBrain: the brain's epigenetic landscape in a snapshot.bioRxiv : the preprint server for biology · 2026Article
- Current progress of labeling strategies in tissue clearing for large-scale biological visualization.Frontiers in cell and developmental biology · 2026Review
- An optimized click chemistry method allows visualization of proliferating neuronal progenitors in the mouse brain.Cell reports methods · 2025Article
- Signal Amplification for Fluorescent Staining of Single Particles in Liquid Biopsies: Circulating Tumour Cells and Extracellular Vesicles.Journal of extracellular vesicles · 2025Article
- Imaging cellular activity simultaneously across all organs of a vertebrate reveals body-wide circuits.bioRxiv : the preprint server for biology · 2025Article
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Authors and funding
13 authors at 4 institutions in 1 country.
Funding
Abstract
For comprehensive studies of the brain structure and function, fluorescence imaging of the whole brain is essential. It requires large-scale volumetric imaging in cellular or molecular resolution, which could be quite challenging. Recent advances in tissue clearing technology (e.g. CLARITY, PACT) provide new solutions by homogenizing the refractive index of the samples to create transparency. However, it has been difficult to acquire high quality results through immunofluorescence (IF) staining on the cleared samples. 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. We show that TSA-PACT is able to reduce the opacity of the zebrafish brain by more than 90% with well-preserved structure. Compared to traditional method, TSA-PACT achieves approximately tenfold signal amplification and twofold improvement in signal-to-noise ratio (SNR). Moreover, both the structure and the fluorescent signal persist for at least 16 months with excellent signal retention ratio. 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.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.