PBS is short for Phosphate Buffered Solution, while HBSS stands for Hank’s Balanced Salt Solution. As the name suggests, both solutions have different ingredients and forms. PBS solution is mostly in liquid form while HBSS is available in both powder and liquid form.
The chemical compounds used to make both solutions vary depending on the manufacturer or availability of the chemical.
When it comes to cell biology or biochemistry, saline is common in both streams. Saline solutions are chemicals required in laboratories to carry out the various tests. PBS and HBSS are the most commonly used saline solutions in any cell biology laboratory.
But what use does this saline solution have? and what are the differences with this solution? Let’s discuss each solution in detail.
PBS Vs HBSS
As we discussed above, these two solutions are the most common solutions in the field of cell biology. But did you know what the main differences are between PBS and HBSS, aside from the name and chemical compound?
The first and most important difference between PBS and HBSS solutions is that PBS solutions are only used in biochemical and molecular biology experiments, while HBSS solutions are used in all biological research. Both solutions are salt based and their main function is to maintain a neutral pH. However, they are made of different materials.
As you know, “PBS” stands for Phosphate Buffer Saline and “HBSS” stands for Hank’s Balanced Salt Solution. These are the two special forms of salt solution to which a special substance or chemical is added to achieve their purpose.
How can you use PBS and HBSS solutions in biological experiments? Pour both solutions into the appropriate Petri dishes, then add the cells or tissue you want to study or use during an activity. The way both solutions are used may look the same, but the roles they play are different.
If you use PBS in your experiment or test, you need to know that PBS is a buffer solution that maintains the neutral pH of the solution and cell osmolarity and protects your test sample from destruction. It is also non-toxic to cells.
But what exactly is a buffer solution? Buffer solution is made from the “weak” acid or base and the corresponding acid or base. Normally, buffer solutions are added to the other solution to maintain the specific pH of that solution.
In addition to the Bffer function of the PBS, it is also used to dilute the materials and wash the containers in which you want to keep the cells or tissue in order to maintain effective hygiene.
On the other hand, if you use Hank’s balanced salt solution, it is an isotonic solution specifically designed to maintain the specific pH and osmotic pressure of the test sample. Its main role is to supply enough salt ions to the test sample to maintain intracellular water content.
While PBS only protects your sample from destruction, HBSS is used to preserve the sample in its natural state and structure. However, adding an inappropriate amount of salt ions in the solution it will cause the cells to shrink or rupture due to the imbalance in osmotic pressure.
The main ingredients in making a PBS solution are water, sodium chloride, as well as sodium phosphate, potassium chloride and potassium phosphate. All these salts are added in a certain amount to make a PBS solution.
On the other hand, if you want to make an HBSS solution, you will need many more ingredients than those used in the PBS solution.
The main ingredients used in the preparation of HBSS solution are potassium chloride, disodium hydrogen phosphate, potassium dihydrogen phosphate, sodium formate, calcium chloride, magnesium chloride and sodium chloride. In addition to this ingredient, glucose is also used to prepare the HBSS solution.
Some other ingredients can also be added to make HBSS, including hydrated magnesium sulfate and sodium bicarbonate.
However, because of these many ingredients, producing an HBSS solution carries risks. Most people buy them instead of making them themselves.
Comparison Table Between PBS And HBSS
| Parameters of Comparison | PBS | HBSS |
| Stands for | Phosphate Buffered Saline | Hank’s Balanced Salt Solution |
| Ingredients | NaCl, KCl, Na2HPO4, KH2PO4. | Inorganic salts, glucose or pyruvate, phenol red, and sodium bicarbonate. |
| Forms | Made in liquid form only. | Can be made in both liquid and powdered form. |
| Storage Temperature | 15-30 Celsius Degrees | 4-8 Celsius Degrees. |
| Applications | washing cells, transporting tissue, diluting cells, and preparing reagents. | Mammalian Cell culture, transport media |
What Is PBS?
PBS or phosphate buffer saline is a water-based buffered saline solution used to maintain the neutral pH of the test sample. The water level and ion concentration of the PBS solution are equivalent to those of the human body, indicating that the PBS solutions are also isotonic, which helps maintain osmotic pressure.
Since PBS solutions are non-toxic to cells, they have many uses including substance dilution and detachment of clumped cells, etc. In recent years, PBS solutions have been in great demand as they are used as an alternative to viral transport media for RNA storage in viruses including SARS-CoV-2 or Coronavirus.
A PBS solution can be prepared by various methods. When the buffer is used on living tissue, the preparation requires potassium chloride or magnesium chloride, while when the buffer is used on fixed tissue, neither potassium chloride nor magnesium chloride is required.
During the preparation, the pH value can be adjusted with hydrochloric acid or sodium hydroxide.
The most common components of a PBS solution are NaCl, KCl, Na2HPO4, and KH2PO4 in precisely calculated amounts. PBS solutions can also be prepared using commercially manufactured PBS-buffered tablets or capsules. PBS solutions are stored either at room temperature or in the refrigerator.
What Is HBSS?
HBSS stands for Hank’s Balanced Salt Solution. This solution was named after biologist John H. Hanks, who formulated it in 1940. This solution is used as a buffering system in cell culture and helps maintain optimal pH or cell growth. The optimum pH values are between 7.0 and 7.4.
Because HBBS solutions contain glucose and sodium bicarbonate, they are used for short-term cell maintenance outside of the growth medium.
These solutions are also used for a variety of cell culture applications such as washing cells, transporting cells, or diluting cells. HBSS solutions formed with calcium and magnesium are used as transport media or to prepare reagents.
An HBSS solution is prepared from a powdered base medium combined with culture-grade tissue water. The usual components of an HBSS solution are inorganic salts, glucose or pyruvate, phenol red, and sodium bicarbonate. After preparation, samples of HBSS solutions are tested to confirm the absence of bacterial or fungal contamination.
HBSS solutions are sensitive and are supplied in gamma-irradiated, sterile bottles. HBSS solutions are stored at a low temperature of 4-8 °C and protected from direct light. They are also more dangerous to manufacture compared to PBS solutions because human cells contain biologically hazardous materials.
Main Difference Between PBS And HBSS
A PBS solution is generally easier to prepare, while the manufacturing process of an HBSS solution can be hazardous and extra precautions are therefore important.
PBS solutions can be stored at a temperature ranging from 15 to 30°C while HBSS solutions are stored at a temperature ranging from 4 to 8°C.
PBS solutions can only be made in liquid form while HBSS solutions can be made in both liquid and powder form.
PBS solutions are pure by default. On the other hand, HBSS solutions must be checked for bacterial or fungal contamination.
PBS solutions are non-toxic to cells while HBSS can cause cell death.