Abstract
A method is presented for measuring intracellular pH (pHi) spectrophotometrically in hippocampal slices using the pH dye indicator, Neutral red (NR). Measurements of pHi by NR were compared directly with the creatine kinase (CK) equilibrium method. Slices were bathed in artificial cerebrospinal fluid buffered with bicarbonate/CO2. Intracellular pH in hippocampal slices was found to be more alkaline (approximately 0.3 pH units) than buffer pH and far more alkaline (0.5 pH units) than extracellular pH. Resting intracellular alkalinity was observed by using both the NR and the CK equilibrium methods. The method may be useful for studies of pH regulation in intact functioning tissues in vitro where rapid and repeated measurements are necessary or where cell size precludes measurements with pH-sensitive microelectrodes.
| Original language | English |
|---|---|
| Pages (from-to) | 25-34 |
| Number of pages | 10 |
| Journal | Journal of Neuroscience Methods |
| Volume | 27 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jan 1 1989 |
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Keywords
- Brain slice
- Creatine kinase equilibrium
- Hydrogen ion
- Neutral red
- pH
- Spectrophotometry
ASJC Scopus subject areas
- Neuroscience(all)
Cite this
Determination of intracellular pH in the in vitro hippocampal slice preparation by transillumination spectrophotometry of Neutral red. / Sick, Thomas; Whittingham, Tim S.; LaManna, Joseph C.
In: Journal of Neuroscience Methods, Vol. 27, No. 1, 01.01.1989, p. 25-34.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Determination of intracellular pH in the in vitro hippocampal slice preparation by transillumination spectrophotometry of Neutral red
AU - Sick, Thomas
AU - Whittingham, Tim S.
AU - LaManna, Joseph C.
PY - 1989/1/1
Y1 - 1989/1/1
N2 - A method is presented for measuring intracellular pH (pHi) spectrophotometrically in hippocampal slices using the pH dye indicator, Neutral red (NR). Measurements of pHi by NR were compared directly with the creatine kinase (CK) equilibrium method. Slices were bathed in artificial cerebrospinal fluid buffered with bicarbonate/CO2. Intracellular pH in hippocampal slices was found to be more alkaline (approximately 0.3 pH units) than buffer pH and far more alkaline (0.5 pH units) than extracellular pH. Resting intracellular alkalinity was observed by using both the NR and the CK equilibrium methods. The method may be useful for studies of pH regulation in intact functioning tissues in vitro where rapid and repeated measurements are necessary or where cell size precludes measurements with pH-sensitive microelectrodes.
AB - A method is presented for measuring intracellular pH (pHi) spectrophotometrically in hippocampal slices using the pH dye indicator, Neutral red (NR). Measurements of pHi by NR were compared directly with the creatine kinase (CK) equilibrium method. Slices were bathed in artificial cerebrospinal fluid buffered with bicarbonate/CO2. Intracellular pH in hippocampal slices was found to be more alkaline (approximately 0.3 pH units) than buffer pH and far more alkaline (0.5 pH units) than extracellular pH. Resting intracellular alkalinity was observed by using both the NR and the CK equilibrium methods. The method may be useful for studies of pH regulation in intact functioning tissues in vitro where rapid and repeated measurements are necessary or where cell size precludes measurements with pH-sensitive microelectrodes.
KW - Brain slice
KW - Creatine kinase equilibrium
KW - Hydrogen ion
KW - Neutral red
KW - pH
KW - Spectrophotometry
UR - http://www.scopus.com/inward/record.url?scp=0024528013&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0024528013&partnerID=8YFLogxK
U2 - 10.1016/0165-0270(89)90050-2
DO - 10.1016/0165-0270(89)90050-2
M3 - Article
C2 - 2918752
AN - SCOPUS:0024528013
VL - 27
SP - 25
EP - 34
JO - Journal of Neuroscience Methods
JF - Journal of Neuroscience Methods
SN - 0165-0270
IS - 1
ER -