Slice Buffer Preparation
Slice Buffer Preparation - Related Videos
02:56
Buffers
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2020
A solution containing appreciable amounts of a weak conjugate acid-base pair is called a buffer solution, or a buffer. Buffer solutions resist a change in pH when small amounts of a strong acid or a strong base are added. A solution of acetic acid and sodium acetate is an example of a buffer that consists of a weak acid and its salt: CH3COOH (aq) + CH3COONa (aq). An example of a buffer that consists of a weak base and its salt is a solution of ammonia and ammonium chloride: NH3 (aq) + NH4Cl...
08:13
Buffers - Instructor Prep
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2020
Source: Smaa Koraym at Johns Hopkins University, MD, USA Preparation of SolutionsExpand Here, we show the laboratory preparation for 10 students working in pairs, with some excess. Please adjust quantities as needed. To set up for this lab experiment, wear the appropriate personal protective equipment, including a lab coat, chemical splash goggles, and gloves. All solutions for this lab should be prepared in a chemical fume hood. Prepare 100 mL of a 1 M NaOH solution. Note:...
04:35
Buffers - Concepts
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2020
Buffers When an Arrhenius acid (HA) is added to water, it dissociates into its conjugate base (A-) and a hydrogen cation (H+). HA + H2O → H+(aq) + A-(aq) The amount of hydrogen ions present in the solution determines the acidity of the solution, where more hydrogen ions indicate a lower or more acidic pH. Similarly, when a strong Arrhenius base (BOH) is added to water, it dissociates into its conjugate acid (B+) and a hydroxide ion (OH-). BOH + H2O → B+(aq) + OH-(aq) Generally, the addition of...
14:31
Buffers - Student Protocol
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2020
Source: Smaa Koraym at Johns Hopkins University, MD, USA Preparing 50 mM NaH2PO4 Buffer, pH 7Expand In the first part of this experiment, you will prepare a sodium phosphate solution buffered at pH 7.0. Monosodium phosphate is a weak acid with the conjugate base, disodium phosphate. Unadjusted monosodium phosphate solutions usually have a pH of about 4 - 6. Buffers are most effective close to their pKa, which is 6.8 to 7.2 for monosodium phosphate. So, you will use NaOH to push the...
08:12
Thin Sectioning of Slice Preparations for Immunohistochemistry
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Cited by 7 •
2007
The present method allows reproducible cryostat sectioning of small, difficult-to-manage, tissue pieces, such as biopsies and brain slices. We utilize a simple aluminum freezing stage to facilitate handling of tissue and a standard cryostat to routinely produce 5-10 micron serial sections from 400 micron thick brain slices.
FAQs
Ionic composition, osmolarity, pH, temperature, and oxygenation must be controlled together rather than treated as independent settings. Their combined balance helps the solution reproduce important features of extracellular fluid while limiting physiological disturbance during tissue removal. Variations in any of these properties can increase stress on neurons and glial cells and reduce the functional quality of the resulting slices.
The buffer limits several stresses that arise when brain tissue is removed and cut. Appropriate ionic conditions and osmolarity help protect cell membranes, while controlled pH, temperature, and oxygenation support tissue stability. These conditions also help limit metabolic stress and excessive excitotoxic activity, preserving neuronal and glial integrity sufficiently for subsequent functional experiments.
After removal from the brain, tissue no longer receives support through its original circulation, so the preparation must maintain conditions that help sustain cellular function. Oxygenation is therefore a central part of Slice Buffer Preparation, alongside temperature and chemical control. Maintaining these conditions during sectioning helps reduce metabolic stress and supports the survival and activity of cells in acute slices.
A preparation workflow starts by formulating the aqueous solution, then checking and controlling its ionic composition, osmolarity, pH, temperature, and oxygenation. The solution should be maintained under the intended conditions while tissue is sectioned, rather than adjusted only afterward. This coordinated handling gives the brain tissue consistent protection throughout the transition from intact tissue to acute slice.
The main outcome is tissue that remains viable and functionally intact after sectioning. Researchers can evaluate whether neurons and glial cells tolerate the preparation well enough for experiments that depend on physiological activity. A suitable buffer supports slices for electrophysiology, calcium imaging, pharmacological studies, and analyses of synaptic transmission or neural circuit function.
Carefully prepared slices provide an experimental system for examining neural function after tissue removal. Electrophysiology can assess cellular or synaptic activity, calcium imaging can follow activity-related signals, and pharmacological studies can test responses to applied compounds. The same preparation also supports investigations of synaptic transmission and neural circuit function, making buffer quality relevant across several neuroscience approaches.