Hi everyone,
I established a murine spheroid for glioblastoma, and I want to fix and stain them to conduct immunofluorescences. I have tried several times, but the size of my spheroids are small, and I cannot see them during Paraffin embedding step. I would be grateful if you could give me some hints on doing this step.
Samir Ranjan Panda
Hi Vahid Khaki
Please find the attached protocols.
Protocol for preparation of blocks from small spheroids.
- Spheroid Processing and Embedding for Histology (Corning): https://www.corning.com/catalog/cls/documents/protocols/CLS-AN-431_DL.pdf: https://www.corning.com/catalog/cls/documents/protocols/CLS-AN-431_DL.pdf
- Histological Processing of Three-Dimensional Cell Cultures (National Cancer Institute): https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3524419/: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3524419/
- Microarray Embedding/Sectioning for Parallel Analysis of 3D Cell Spheroids (PMC): https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6872729/: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6872729/
Staining Protocol
- Immunofluorescent staining of cancer spheroids and fine-needle aspiration-derived organoids https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5709388/
- Quantitative three-dimensional evaluation of immunofluorescence staining for large whole mount spheroids with light sheet microscopy https://pubmed.ncbi.nlm.nih.gov/34136836/
- Fluorescence-Based Quantitative and Spatial Analysis of Tumour Spheroids: A Proposed Tool to Predict Patient-Specific Therapy Response https://pubmed.ncbi.nlm.nih.gov/34136836/
Thanks,
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