In research in pathology, specimens of cells and tissues are observed to reveal the development and progression of a disease. Various methods have been established to visualize target tissues, cells, and molecules (proteins, nucleic acids, etc.) for observation, including staining with dyes, immunostaining, and in situ hybridization. Fujifilm Wako offers a wide line-up of reagents necessary for various processes of pathological research.
Processes from Specimen Preparation to Microscopic Observation
Sample processing varies depending on the cells or tissues to be observed and the method of staining. The general processes from paraffin section preparation to observation by light microscope involve the following steps:
- 1. Dissection
Target tissue is excised from the animal. Perfusion fixation may be necessary depending on the tissue to be observed.
- 2. Fixation
Molecular (mainly proteins) in the tissue are immobilized to keep the condition of the specimen as close to that of the living tissue as possible for an extended period of time.
Available reagents: Fixatives
- 3. Decalcification and defatting (depending on the sample)
Hard tissues such as bone are decalcified, and lipid-rich tissues such as brain, skin, or adipose tissue are defatted.
Available reagents: Tissue Decalcifying Solutions
- 4. Dehydration
For paraffin embedding, the water in the tissue is removed with alcohol, and then the alcohol is replaced with a clearing agent miscible with paraffin.
Available reagents: Clearing Agents
- 5. Embedding
Soft tissues are difficult to cut into thin sections as they are. Embedding tissues in paraffin or other material makes the tissue sufficiently hard.
Available reagents: Tissue Embedding Agents
- 6. Sectioning
Embedded specimens are cut into thin sections for staining and observation.
- 7. Staining
Thin sections are stained with dyes or labeled with antibodies and probes. Pretreatment such as deparaffinization, antigen retrieval, and blocking may be performed. It is also possible to decolorize stained specimens with a decolorizing reagent and apply another stain.
Available reagents: Antigen Retrieval Reagents / Tissue Decolorizing Reagents
- 8. Mounting
A mounting agent is added to the glass slides to fix and preserve specimens on the slides for long-term storage. Dehydration may be performed when a solvent-based mounting medium is used.
- 9. Observation
Prepared specimens are observed under a microscope.
- Yamada, K.: “Histochemistry”, Nankodo, Japan, (1987). (Japanese)
- “Handbook for Staining and Bioimaging Experiments 5th ed.” ed. by Takata, K., Saito, N. and Kawakami, H., Yodosha, Japan, (2012). (Japanese)
- Okubo, K.: “Master of cell and tissue staining”, Yodosha, Japan, (2018). (Japanese)
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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