Protocol for hbss

Written by deborah farson
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Protocol for hbss
Cells growing on plates and flasks in an incubator (culture plates and flasks image by Fotocie from

Hank's buffered salt solution (HBSS), also known as Hank's buffer, is widely used in laboratories to maintain cultured cells in a healthy state. The buffer keeps the cells from becoming too acidic or basic and provides essential salts.

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John H. Hanks developed this buffer while at Johns Hopkins University and called it Buffered Saline Solution (BSS). The buffer became famous in 1949, when it was used by scientists to help cells in culture grow the polio virus for the first time.


The term "pH" is used to describe how basic or acidic a solution is on a scale of 1-12, with 1 being the most acidic, 12 the most basic and 7 neutral. A buffer is a mixture of acids and bases that have the capacity to absorb more acids or bases without disturbing the pH. The human blood is buffered to maintain a pH around 7.35.


The buffer contains a specific ratio of sodium chloride, potassium chloride, sodium hydrogen phosphate, potassium dihydrogen phosphate, calcium chloride, magnesium chloride and sodium bicarbonate. Glucose -- sugar -- may also be added. This buffer can be made in the laboratory or purchased commercially. It is usually stored at room temperature.

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