What is the Difference Between HBSS and PBS

The main difference between HBSS and PBS is that HBSS contains various salts, glucose, and buffers to maintain physiological pH, while PBS contains phosphate ions, chloride ions, and sodium ions.

HBSS (Hank’s Balanced Salt Solution) and PBS (Phosphate Buffered Saline) are both types of buffer solutions commonly used in biological and biochemical research.

Key Areas Covered

1. What is HBSS  
      – Definition, Features, Applications
2. What is PBS
      – Definition, Features, Applications 
3. Similarities Between HBSS and PBS
      – Outline of Common Features
4. Difference Between HBSS and PBS
      – Comparison of Key Differences
5. FAQ: HBSS and PBS
      – Frequently Asked Questions

Key Terms

Hank’s Balanced Salt Solution, HBSS, PBS, Phosphate Buffered Saline

Difference Between HBSS and PBS - Comparison Summary

What is HBSS

Hank’s Balanced Salt Solution (HBSS) is a crucial component in various biological and biomedical applications, particularly in cell culture and experimental settings. Its composition is meticulously designed to mimic the physiological conditions of living organisms, providing an isotonic and buffered environment for cells.

The primary constituents of HBSS include salts such as sodium chloride, potassium chloride, calcium chloride, magnesium sulfate, sodium phosphate, and sodium bicarbonate. These components maintain osmotic balance, ensuring that the concentration of solutes inside and outside the cells is similar. This isotonic nature prevents cell swelling or shrinkage, maintaining cell integrity during experiments or storage.

Calcium and magnesium ions in HBSS play essential roles in various cellular processes. Calcium, for instance, is a critical signaling molecule involved in processes like cell adhesion, migration, and muscle contraction. Magnesium is crucial for stabilizing nucleic acids and participating in enzymatic reactions. The precise concentration of these ions in

HBSS is carefully formulated to support cellular functions without inducing unwanted effects. Furthermore, the buffering capacity of HBSS, often achieved through the inclusion of sodium bicarbonate, helps maintain a stable pH level.

HBSS is commonly used for washing and maintaining cells during experimental procedures. Its ability to preserve cell viability and functionality makes it a preferred choice for procedures like cell dissociation, washing cells before analysis, or diluting samples. Moreover, the absence of proteins and other biological molecules in its formulation reduces the risk of interference with experimental outcomes.

Differentiate HBSS vs PBS

In addition to its use in cell culture, HBSS is useful in techniques such as fluorescence-activated cell sorting (FACS) and flow cytometry. The balanced salt solution provides a neutral environment for these sensitive analytical methods, preventing alterations in cell properties that could skew results.

What is PBS

PBS (Phosphate Buffered Saline) solution is a solution made of phosphate ions, sodium chloride, and sometimes potassium chloride, and serves as a versatile buffer with a pH close to the physiological range, typically around 7.4. This pH stability is crucial for maintaining the integrity and functionality of biological molecules.

The phosphate component in PBS plays a pivotal role in buffering. Phosphates can exist in multiple ionization states, allowing them to act as a weak acid or base depending on the solution’s pH. This property helps PBS resist drastic changes in acidity or alkalinity, providing a stable environment for delicate biological processes. Moreover, the choice of sodium and potassium salts contributes to the osmotic balance within the solution, mimicking the physiological conditions essential for cellular studies.

The concentration of salts in PBS is carefully calibrated to mirror the ionic composition of the human body. Sodium chloride, a common salt, mirrors the concentration found in extracellular fluids, ensuring that cells immersed in PBS experience minimal osmotic stress. In fact, the inclusion of potassium chloride, which is prevalent in intracellular fluids, helps maintain a balance that supports the functionality of cellular components.

PBS is frequently employed in various laboratory procedures, such as cell culture, molecular biology, and immunohistochemistry. In cell culture, PBS is used for washing and rehydrating cells without introducing foreign ions that might interfere with experiments. In molecular biology, it provides an ideal medium for enzymatic reactions, as its buffering capacity stabilizes pH, ensuring optimal conditions for enzymes. Additionally, PBS is useful in immunohistochemistry to prepare tissues for analysis, offering a gentle yet effective means of preserving the structural and antigenic integrity of biological specimens.

Similarities Between HBSS and PBS

  • Both are isotonic solutions commonly used in biological and medical research.
  • Moreover, they are designed to maintain a stable pH. 

Difference Between HBSS and PBS


HBSS is a balanced salt solution that closely mimics the electrolyte composition of the extracellular fluid in the body. PBS, on the other hand, is a buffer solution containing phosphate salts. 


HBSS contains various salts, glucose, and buffering agents, while PBS contains phosphate salts, sodium chloride, and sometimes potassium chloride.


While HBSS typically has a pH of around 7.2 to 7.4, PBS usually has a pH of around 7.4.


HBSS is commonly used in cell culture, tissue dissociation, and maintaining cell viability during experiments, whereas PBS is used for various biological and biochemical applications, including cell washing, dilution, and as a buffer for immunohistochemistry.


What is HBSS used for?

HBSS is used for washing cells before dissociation, transporting cells, or diluting cells.

Is PBS or HBSS better for cell culture?

PBS is versatile and suitable for many applications, especially when live cells are not involved. HBSS, with added components like glucose, is preferred for experiments with live cells to support their metabolic needs.

Can PBS be used instead of HBSS?

In many cases, PBS (Phosphate Buffered Saline) can be used as a substitute for HBSS (Hank’s Balanced Salt Solution).


In conclusion, the main difference between HBSS and PBS is their content. HBSS contains various salts, glucose, and buffers to maintain physiological pH, while PBS contains phosphate ions, chloride ions, and sodium ions.


1. “HBSS.” Wikipedia. Wikipedia Foundation. 
2. “Phosphate-Buffered Saline” Wikipedia. Wikipedia Foundation. 

Image Courtesy:

1. “Laboratory glassware with colored liquids on light background”  By Marco Verch Professional Photography (CC BY 2.0 DEED) via Flickr

About the Author: Hasini A

Hasini is a graduate of Applied Science with a strong background in forestry, environmental science, chemistry, and management science. She is an amateur photographer with a keen interest in exploring the wonders of nature and science.

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