Determination of intracellular pH in the in vitro hippocampal slice preparation by transillumination spectrophotometry of Neutral red

Thomas Sick, Tim S. Whittingham, Joseph C. LaManna

Research output: Contribution to journalArticle

11 Citations (Scopus)

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 languageEnglish
Pages (from-to)25-34
Number of pages10
JournalJournal of Neuroscience Methods
Volume27
Issue number1
DOIs
StatePublished - Jan 1 1989

Fingerprint

Transillumination
Neutral Red
Spectrophotometry
Creatine Kinase
In Vitro Techniques
Microelectrodes
Bicarbonates
Cell Size
Cerebrospinal Fluid
Buffers
Coloring Agents

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 journalArticle

@article{1c05ac97664c4738890bbf61efe8b381,
title = "Determination of intracellular pH in the in vitro hippocampal slice preparation by transillumination spectrophotometry of Neutral red",
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.",
keywords = "Brain slice, Creatine kinase equilibrium, Hydrogen ion, Neutral red, pH, Spectrophotometry",
author = "Thomas Sick and Whittingham, {Tim S.} and LaManna, {Joseph C.}",
year = "1989",
month = "1",
day = "1",
doi = "10.1016/0165-0270(89)90050-2",
language = "English",
volume = "27",
pages = "25--34",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",
number = "1",

}

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 -