ArticleDevelopmental biology2025
Comparative analysis of fixation techniques for signal detection in avian embryos.
Article in Developmental biology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed.
- Advances in Understanding Calcium Sensitization in Gastrointestinal Smooth Muscles: Pathways, Interstitial Cells, and Disease.Journal of neurogastroenterology and motility · 2026Review
- A spatial and temporal atlas of tubulin isotype expression during neural crest EMT.bioRxiv : the preprint server for biology · 2026Article
- A molecular and spatial resource defining tubulin isotype organization during corneal development.bioRxiv : the preprint server for biology · 2026Article
- Fixation matters: duration in fixative prior to immunofluorescent analysis directly impacts macrophage visualization in epithelial tissues.Discovery immunology · 2026Article
- The Protein Kinase aPKC as Well as the Small GTPases RhoA and Cdc42 Regulates Neutrophil Chemotaxis Partly by Recruiting the ROCK Kinase to the Leading Edge.Genes to cells : devoted to molecular & cellular mechanisms · 2025Article
- Spatiotemporal characterization of cyclooxygenase pathway enzymes during vertebrate embryonic development.Developmental biology · 2025Article
- Characterization and neurogenic responses of primary and immortalized Müller glia.Frontiers in cell and developmental biology · 2025Article
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Authors and funding
3 authors.
Funding
Abstract
The choice of fixation method significantly impacts tissue morphology and visualization of gene expression and proteins after in situ hybridization chain reaction (HCR) or immunohistochemistry (IHC), respectively. In this study, we compared the effects of paraformaldehyde (PFA) and trichloroacetic acid (TCA) fixation techniques prior to HCR and IHC on chicken embryos. Our findings underscore the importance of optimizing fixation methods for accurate visualization and subsequent interpretation of HCR and IHC results, with implications for probe and antibody validation and tissue-specific protein localization studies. We found that TCA fixation resulted in larger and more circular nuclei and neural tubes compared to PFA fixation. Additionally, TCA fixation altered the subcellular fluorescence signal intensity of various proteins, including transcription factors, cytoskeletal proteins, and cadherins. Notably, TCA fixation revealed protein signals in tissues that may be inaccessible with PFA fixation. In contrast, TCA fixation proved ineffective for mRNA visualization. These results highlight the need for optimization of fixation protocols depending on the target and model system, emphasizing the importance of methodological considerations in biological analyses.
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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 Socratic graph.