What is Hank's Balanced Salt Solution (HBSS)?
Hank's Balanced Salt Solution (HBSS) is a specially formulated buffer solution that is widely used in
cell culture and histology. It provides essential inorganic ions and maintains the osmotic balance, pH, and other physiological conditions necessary for the survival of cells outside their natural environment.
Components of HBSS
HBSS typically contains a balanced mixture of
salts, glucose, and sometimes phenol red as a pH indicator. Key components include sodium chloride, potassium chloride, calcium chloride, magnesium sulfate, sodium bicarbonate, and glucose. These components collectively simulate the ionic composition of the human body fluids, creating an ideal environment for cells.
1. Cell Viability: HBSS helps to maintain cell viability during the preparation and processing of histological specimens.
2. Tissue Preservation: It is used to keep tissues hydrated and prevent them from drying out, which is crucial for accurate histological analysis.
3. Transport Medium: HBSS serves as an excellent medium for transporting tissues from the surgical suite to the histology lab without compromising cell integrity.
How is HBSS Prepared?
To prepare HBSS, specific quantities of its components are dissolved in distilled water, and the pH is adjusted to around 7.2-7.4. The solution is then sterilized, typically through filtration, to ensure it is free from contaminants. Commercially available HBSS can be purchased in ready-to-use formats, saving time and ensuring consistency.
Applications of HBSS in Histology
HBSS is used in various stages of histological workflows:1. Sample Collection: During biopsy or surgery, tissues are often placed in HBSS to maintain cellular integrity until they can be processed.
2. Rinsing and Washing: HBSS is used to rinse tissues and cells to remove blood, debris, and other contaminants.
3. Enzyme Reactions: It provides a suitable environment for enzyme reactions during tissue processing, such as digestion or dissociation of tissues.
4. Cryopreservation: HBSS can be used as a base solution for cryopreserving cells and tissues, ensuring they remain viable when thawed for later analysis.
HBSS vs. Other Solutions
Compared to other solutions, such as
PBS (Phosphate Buffered Saline) or
DPBS (Dulbecco's Phosphate Buffered Saline), HBSS is often preferred for specific applications that require the presence of calcium and magnesium ions, which are necessary for certain cellular functions. However, PBS and DPBS are also widely used in histology, particularly for washing and diluting cells.
Considerations When Using HBSS
When using HBSS, it is essential to consider the following:1. Sterility: Ensure that HBSS is sterile to prevent contamination of tissue samples.
2. Component Concentrations: Use the correct formulation of HBSS (with or without calcium and magnesium) depending on the specific application.
3. Storage: Store HBSS at the recommended temperature, usually 2-8°C, to maintain its integrity.
Conclusion
HBSS is a critical tool in histology, providing a stable environment for cells and tissues during various stages of processing and analysis. Its balanced ionic composition and ability to maintain physiological conditions make it indispensable for preserving cell viability and integrity. By understanding its components, applications, and proper usage, histologists can ensure the accuracy and reliability of their tissue analyses.
