A novel fluorescence ratiometric method confirms the low solvent viscosity of the cytoplasm

K. Luby-Phelps, S. Mujumdar, R. B. Mujumdar, L. A. Ernst, W. Galbraith, A. S. Waggoner

Research output: Contribution to journalArticle

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Abstract

Two homologous indocyanine dyes, Cy3.18 and Cy5.18, can be used as a ratio pair for fluorometric determination of solvent viscosity. Succinimidyl ester derivatives of these dyes can be attached to inert carrier macromolecules, such as Ficoll 70, for measurement of intracellular or intravesicular solvent viscosity. When the viscosity of the solvent was varied by various methods, the fluorescence intensity ratio (Cy3/Cy5) in a mixture of Cy3.18-Ficoll 70 (Cy3F70) and Cy5.18-Ficoll 70 (Cy5F70) in solution was found to be solely a function of solvent viscosity and was insensitive to other solvent parameters such as dielectric constant, temperature, and the ability of the solvent to form hydrogen bonds. Most important, it was insensitive to the presence of large macromolecules, such as proteins, which increase the shear viscosity but have little effect on solvent viscosity. Following microinjection into the cytoplasm of living tissue culture cells, no binding of Cy3F70 or Cy5F70 to intracellular components was detected by fluorescence recovery after photobleaching. Fluorescence intensity ratio imaging of Cy3F70 and Cy5F70 in nonmotile interphase CV1 and PtK1 cells showed that the solvent viscosity of cytoplasm was not significantly different from water and showed no spatial variation.

Original languageEnglish (US)
Pages (from-to)236-242
Number of pages7
JournalBiophysical Journal
Volume65
Issue number1
StatePublished - 1993

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Ficoll
Viscosity
Cytoplasm
Fluorescence
Coloring Agents
Fluorescence Recovery After Photobleaching
Interphase
Microinjections
Hydrogen
Esters
Cell Culture Techniques
Temperature
Water

ASJC Scopus subject areas

  • Biophysics

Cite this

Luby-Phelps, K., Mujumdar, S., Mujumdar, R. B., Ernst, L. A., Galbraith, W., & Waggoner, A. S. (1993). A novel fluorescence ratiometric method confirms the low solvent viscosity of the cytoplasm. Biophysical Journal, 65(1), 236-242.

A novel fluorescence ratiometric method confirms the low solvent viscosity of the cytoplasm. / Luby-Phelps, K.; Mujumdar, S.; Mujumdar, R. B.; Ernst, L. A.; Galbraith, W.; Waggoner, A. S.

In: Biophysical Journal, Vol. 65, No. 1, 1993, p. 236-242.

Research output: Contribution to journalArticle

Luby-Phelps, K, Mujumdar, S, Mujumdar, RB, Ernst, LA, Galbraith, W & Waggoner, AS 1993, 'A novel fluorescence ratiometric method confirms the low solvent viscosity of the cytoplasm', Biophysical Journal, vol. 65, no. 1, pp. 236-242.
Luby-Phelps K, Mujumdar S, Mujumdar RB, Ernst LA, Galbraith W, Waggoner AS. A novel fluorescence ratiometric method confirms the low solvent viscosity of the cytoplasm. Biophysical Journal. 1993;65(1):236-242.
Luby-Phelps, K. ; Mujumdar, S. ; Mujumdar, R. B. ; Ernst, L. A. ; Galbraith, W. ; Waggoner, A. S. / A novel fluorescence ratiometric method confirms the low solvent viscosity of the cytoplasm. In: Biophysical Journal. 1993 ; Vol. 65, No. 1. pp. 236-242.
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