ArticleThe journal of histochemistry and cytochemistry : official journal of the Histochemistry Society2016
An Improved Immunostaining and Imaging Methodology to Determine Cell and Protein Distributions within the Bone Environment.
Article in The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 11 citations in OpenAlex.
- Oxidative stress and impaired insulin secretion in cystic fibrosis pig pancreas.Advances in redox research · 2022Article
- Silicon Oxynitrophosphide Nanoscale Coating Enhances Antioxidant Marker-Induced Angiogenesis During in vivo Cranial Bone-Defect Healing.JBMR plus · 2021Article
- Effect of rhBMP-2 applied with a 3D-printed titanium implant on new bone formation in rabbit calvarium.Journal of applied oral science : revista FOB · 2021Article
- Bone Phenotyping Approaches in Human, Mice and Zebrafish - Expert Overview of the EU Cost Action GEMSTONE ("GEnomics of MusculoSkeletal traits TranslatiOnal NEtwork").Frontiers in endocrinology · 2021Review
- Aberrant BMP2 Signaling in Patients Diagnosed with Osteoporosis.International journal of molecular sciences · 2020Article
- An Improved Methodology to Evaluate Cell and Molecular Signals in the Reparative Callus During Fracture Healing.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2020Article
- Tissue Preparation and Immunostaining of Mouse Craniofacial Tissues and Undecalcified Bone.Journal of visualized experiments : JoVE · 2019Article
- Confocal/two-photon microscopy in studying colonisation of cancer cells in bone using xenograft mouse models.BoneKEy reports · 2016Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Bone is a dynamic tissue that undergoes multiple changes throughout its lifetime. Its maintenance requires a tight regulation between the cells embedded within the bone matrix, and an imbalance among these cells may lead to bone diseases such as osteoporosis. Identifying cell populations and their proteins within bone is necessary for understanding bone biology. Immunolabeling is one approach used to visualize proteins in tissues. Efficient immunolabeling of bone samples often requires decalcification, which may lead to changes in the structural morphology of the bone. Recently, methyl-methacrylate embedding of non-decalcified tissue followed by heat-induced antigen retrieval has been used to process bone sections for immunolabeling. However, this technique is applicable for bone slices below 50-µm thickness while fixed on slides. Additionally, enhancing epitope exposure for immunolabeling is still a challenge. Moreover, imaging bone cells within the bone environment using standard confocal microscopy is difficult. Here we demonstrate for the first time an improved methodology for immunolabeling non-decalcified bone using a testicular hyaluronidase enzyme-based antigen retrieval technique followed by two-photon fluorescence laser microscopy (TPLM) imaging. This procedure allowed us to image key intracellular proteins in bone cells while preserving the structural morphology of the cells and the bone.
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Registered trials
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