Collection, Embedding and Special Staining of Liver Samples

Collection, Embedding and Special Staining of Liver Samples


The liver, a vital organ in the human body, plays a crucial role in various physiological processes such as metabolism, detoxification, and digestion. Understanding the microscopic structure of the liver is essential for diagnosing and studying various liver diseases. This article aims to delve into the process of collecting liver samples, embedding them for microscopic analysis, and employing special staining techniques to enhance visualization.

Collection of Liver Samples

Before delving into the microscopic examination of liver tissue, it is imperative to collect high-quality samples. Liver biopsy, the most common method for obtaining liver tissue, involves the insertion of a thin needle into the liver to extract a small piece of tissue. Alternatively, during surgery, larger liver samples can be collected for analysis.

When performing a liver biopsy, physicians use imaging techniques such as ultrasound to guide the needle to the desired location within the liver. This ensures precise sampling and minimizes the risk of complications. Once the tissue sample is obtained, it is immediately placed in a fixative solution, such as formalin, to preserve its structure and prevent decay.

Embedding of Liver Samples

After collection, liver tissue samples undergo a series of processing steps to prepare them for microscopic analysis. The first step involves dehydration, where the tissue is gradually immersed in a series of alcohol solutions of increasing concentration to remove water from the tissue while maintaining its structural integrity.

Following dehydration, the tissue is infiltrated with a clearing agent such as xylene to remove the alcohol and make the tissue transparent. This step is crucial for subsequent embedding, as it allows for the infiltration of embedding medium into the tissue.

Once cleared, the tissue is embedded in a solid medium to provide support and facilitate thin sectioning for microscopic examination. Paraffin wax is the most commonly used embedding medium due to its compatibility with both tissue structure and staining techniques. The tissue is immersed in molten paraffin wax, allowing it to impregnate the tissue's spaces and solidify, forming a block that encases the tissue.

Special Staining Techniques

While routine hematoxylin and eosin (H&E) staining provides basic visualization of liver tissue architecture, special staining techniques are employed to highlight specific components or structures within the tissue. These techniques provide additional information that may aid in the diagnosis of certain liver diseases.

Typical scored immunohistochemical staining of liver tissue specimens.Figure 1. Typical scored immunohistochemical staining of liver tissue specimens. (Wang Q, et al.; 2011)

One such technique is Masson's trichrome staining, which is used to visualize collagen fibers within the liver tissue. Collagen deposition is a hallmark of fibrosis, a common feature of chronic liver diseases such as cirrhosis. Masson's trichrome staining differentiates collagen fibers (blue) from other tissue components, allowing for the assessment of fibrotic changes in the liver.

Another important staining technique is Periodic Acid-Schiff (PAS) staining, which highlights glycogen and glycoproteins within the liver tissue. Glycogen accumulation is a characteristic feature of certain liver disorders, such as glycogen storage diseases. PAS staining allows for the detection of glycogen within hepatocytes, aiding in the diagnosis of these conditions.

Additionally, immunohistochemistry (IHC) staining is used to detect specific proteins or antigens within the liver tissue. This technique involves the use of antibodies that bind to target proteins, which are then visualized using chromogenic or fluorescent markers. IHC staining can be used to assess the expression of specific markers associated with liver diseases, such as alpha-fetoprotein in hepatocellular carcinoma.


The microscopic examination of liver tissue plays a crucial role in the diagnosis and study of various liver diseases. Collection of high-quality liver samples, proper embedding techniques, and utilization of special staining methods are essential steps in the histological analysis of liver tissue. By employing these techniques, researchers and clinicians can gain valuable insights into the structural and pathological changes occurring in the liver, ultimately leading to improved diagnosis and treatment of liver diseases.

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  1. Wang Q, et al.; Zinc finger protein ZBTB20 expression is increased in hepatocellular carcinoma and associated with poor prognosis. BMC Cancer. 2011, 11:271.

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