Artificial cerebrospinal fluid

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Artificial cerebrospinal fluid (aCSF) is a buffer solution that is used experimentally to immerse isolated brains, brain slices, or exposed brain regions to supply oxygen, maintain osmolarity, and to buffer pH at biological levels. ACSF is commonly used for electrophysiology experiments to maintain the neurons that are being studied.

Composition

One protocol for electrophysiology recording suggests the following composition for aCSF, with the pH and oxygen level stabilized by bubbling with carbogen (95% O2 and 5% CO2): [1]

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3
.

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Soil pH measure of the acidity or alkalinity in soils

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2
O
, a pale greenish-yellow solid which is not explosive and is stable if kept refrigerated.

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Sodium thiosulfate chemical compound

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Phosphate-buffered saline is a buffer solution commonly used in biological research. It is a water-based salt solution containing disodium hydrogen phosphate, sodium chloride and, in some formulations, potassium chloride and potassium dihydrogen phosphate. The buffer helps to maintain a constant pH. The osmolarity and ion concentrations of the solutions match those of the human body (isotonic).

TAE buffer is a buffer solution containing a mixture of Tris base, acetic acid and EDTA.

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Gibbs–Donnan effect

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Tris-buffered saline (TBS) is a buffer used in some biochemical techniques to maintain the pH within a relatively narrow range. Tris has a slightly alkaline buffering capacity in the 7–9.2 range. The conjugate acid of Tris has a pKa of 8.07 at 25 °C. The pKa declines approximately 0.03 units per degree Celsius rise in temperature. This can lead to relatively dramatic pH shifts when there are shifts in solution temperature. Sodium chloride concentration may vary from 100 to 200 mM, tris concentration from 5 to 100 mM and pH from 7.2 to 8.0. A common formulation of TBS is 150 mM NaCl, 50 mM Tris-HCl, pH 7.6. TBS can also be prepared by using commercially made TBS buffer tablets or pouches.

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A balanced salt solution (BSS) is a solution made to a physiological pH and isotonic salt concentration. Solutions most commonly include sodium, potassium, calcium, magnesium, and chloride. Balanced salt solutions are used for washing tissues and cells and are usually combined with other agents to treat the tissues and cells. They provide the cells with water and inorganic ions, while maintaining a physiological pH and osmotic pressure.

Slice preparation

The slice preparation or brain slice is a laboratory technique in electrophysiology that allows the study of a synapse or neural circuit in isolation from the rest of the brain, in controlled physiological conditions. Brain tissue is initially sliced via a tissue slicer then immersed in artificial cerebrospinal fluid (aCSF) for stimulation and/or recording. The technique allows for greater experimental control, through elimination of the effects of the rest of the brain on the circuit of interest, careful control of the physiological conditions through perfusion of substrates through the incubation fluid, to precise manipulation of neurotransmitter activity through perfusion of agonists and antagonists. However, the increase in control comes with a decrease in the ease with which the results can be applied to the whole neural system.

References

  1. "Brain slice electrophysiology video protocol". abcam. Retrieved 28 March 2017.