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Determination of intracellular pH and compartmentation using diffusion-weighted NMR spectroscopy with pH-sensitive indicators

✍ Scribed by Jack S. Cohen; Menachem Motiei; Shani Carmi; Dana Shiperto; Odeya Yefet; Israel Ringel


Publisher
John Wiley and Sons
Year
2004
Tongue
English
Weight
102 KB
Volume
51
Category
Article
ISSN
0740-3194

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✦ Synopsis


Abstract

The intracellular pH (pHi) of a series of cancer cell lines was determined using the pH‐sensitive indicators imidazole (Im) or histidine (His) and diffusion‐weighted (DW) proton NMR spectroscopy. The DW method allows the observation at high magnetic field gradient values of only the slow‐moving (intracellular) components, thus ensuring complete separation between intra‐ and extracellular components. Using the chemical shift difference (Δδ) between the imidazole ring C2‐H and C4(5)‐H peaks, we were able to measure the pHi independently of chemical shift standardization. With His, the cell lines gave pHi values of ∼6.5–7.0, whereas with Im, a second, more acidic compartment (pHi = 5.5–5.8) was also observed. An inverse correlation was also found between pHi and the intracellular lactate concentration. This method may be applicable to in vivo pH determinations. Magn Reson Med 51:900–903, 2004. © 2004 Wiley‐Liss, Inc.