Time-domain fluorescence lifetime imaging for intracellular pH sensing in living tissues

AbstractpH sensing in living cells represents one of the most prominent topics in biochemistry and physiology. In this study we performed one-photon and two-photon time-domain fluorescence lifetime imaging with a laser-scanning microscope using the time-correlated single-photon counting technique for imaging intracellular pH levels. The suitability of different commercial fluorescence dyes for lifetime-based pH sensing is discussed on the basis of in vitro as well of in situ measurements. Although the tested dyes are suitable for intensity-based ratiometric measurements, for lifetime-based techniques in the time-domain so far only BCECF seems to meet the requirements of reliable intracellular pH recordings in living cells.

[1]  Alan G. Ryder,et al.  A Compact Violet Diode Laser-Based Fluorescence Lifetime Microscope , 2002, Journal of Fluorescence.

[2]  Sytsma,et al.  Time‐gated fluorescence lifetime imaging and microvolume spectroscopy using two‐photon excitation , 1998 .

[3]  B. Walz,et al.  Dopamine-induced epithelial K(+) and Na(+) movements in the salivary ducts of Periplaneta americana. , 2001, Journal of insect physiology.

[4]  W. Boron,et al.  Regulation of intracellular pH. , 2004, Advances in physiology education.

[5]  A. Baumann,et al.  The aminergic control of cockroach salivary glands. , 2006, Archives of insect biochemistry and physiology.

[6]  K. Svoboda,et al.  Principles of Two-Photon Excitation Microscopy and Its Applications to Neuroscience , 2006, Neuron.

[7]  Y. K. Levine,et al.  Quantitative pH imaging in cells using confocal fluorescence lifetime imaging microscopy. , 1995, Analytical biochemistry.

[8]  Borivoj Vojnovic,et al.  Monitoring conformational changes of proteins in cells by fluorescence lifetime imaging microscopy. , 2003, The Biochemical journal.

[9]  W. Denk,et al.  Two-photon laser scanning fluorescence microscopy. , 1990, Science.

[10]  A Kusumi,et al.  Comparison of two‐photon excitation laser scanning microscopy with UV‐confocal laser scanning microscopy in three‐dimensional calcium imaging using the fluorescence indicator Indo‐1 , 1997, Journal of microscopy.

[11]  B. Lindemann,et al.  Multi-photon microscopy of cell types in the viable taste disk of the frog , 2003, Cell and Tissue Research.

[12]  S. Frings,et al.  Chloride Accumulation in Mammalian Olfactory Sensory Neurons , 2004, The Journal of Neuroscience.

[13]  Henryk Szmacinski,et al.  Fluorescence lifetime-based sensing and imaging. , 1995, Sensors and actuators. B, Chemical.

[14]  A Liljeborg,et al.  Characterization of probe binding and comparison of its influence on fluorescence lifetime of two pH-sensitive benzo[c]xanthene dyes using intensity-modulated multiple-wavelength scanning technique. , 2000, Analytical biochemistry.

[15]  T. Wensel,et al.  Nanosecond fluorescence microscopy. Emission kinetics of fura-2 in single cells. , 1991, Biophysical journal.

[16]  Hans C. Gerritsen,et al.  Fluorescence lifetime imaging using a confocal laser scanning microscope , 1992 .

[17]  R. Tsien,et al.  A new generation of Ca2+ indicators with greatly improved fluorescence properties. , 1985, The Journal of biological chemistry.

[18]  B. Walz,et al.  Salivary glands of the cockroach, Periplaneta americana: New data from light and electron microscopy , 1994, Journal of morphology.

[19]  K. König,et al.  Fluorescence lifetime imaging by time‐correlated single‐photon counting , 2004, Microscopy research and technique.

[20]  P. Hegyi,et al.  Measurement of Intracellular pH in Pancreatic Duct Cells: A New Method for Calibrating the Fluorescence Data , 2004, Pancreas.

[21]  P. French,et al.  Imaging fluorescence lifetime heterogeneity applied to GFP‐tagged MHC protein at an immunological synapse * , 2005, Journal of microscopy.

[22]  Hermann Marsoner,et al.  Fluorimetric analysis , 1983 .

[23]  T Wilson,et al.  Low‐cost, frequency‐domain, fluorescence lifetime confocal microscopy , 2004, Journal of microscopy.

[24]  M. Rubart,et al.  Two-photon microscopy of cells and tissue. , 2004, Circulation research.

[25]  J. Lichtman,et al.  Optical sectioning microscopy , 2005, Nature Methods.

[26]  C. Hille,et al.  A vacuolar-type H+-ATPase and a Na+/H+ exchanger contribute to intracellular pH regulation in cockroach salivary ducts , 2007, Journal of Experimental Biology.

[27]  Paul Urayama,et al.  A UV–Visible–NIR fluorescence lifetime imaging microscope for laser-based biological sensing with picosecond resolution , 2003 .

[28]  C G Morgan,et al.  Direct modulation of the effective sensitivity of a CCD detector: a new approach to time‐resolved fluorescence imaging , 2002, Journal of microscopy.

[29]  W. Denk,et al.  Deep tissue two-photon microscopy , 2005, Nature Methods.

[30]  Petr Herman,et al.  Fluorescence lifetime‐resolved pH imaging of living cells , 2003, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[31]  P. French,et al.  High speed optically sectioned fluorescence lifetime imaging permits study of live cell signaling events. , 2007, Optics express.

[32]  N. Marcotte,et al.  Carboxy SNARF-4F as a fluorescent pH probe for ensemble and fluorescence correlation spectroscopies. , 2005, The journal of physical chemistry. B.

[33]  A. Draaijer,et al.  Fluorescence lifetime imaging of oxygen in living cells , 2007, Journal of Fluorescence.

[34]  W. Webb,et al.  Multiphoton fluorescence excitation: new spectral windows for biological nonlinear microscopy. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[35]  P J Tadrous,et al.  Methods for imaging the structure and function of living tissues and cells: 2. Fluorescence lifetime imaging , 2000, The Journal of pathology.

[36]  Walz,et al.  The effects of serotonin and dopamine on salivary secretion by isolated cockroach salivary glands , 1996, The Journal of experimental biology.

[37]  A Liljeborg,et al.  Confocal pH imaging of microscopic specimens using fluorescence lifetimes and phase fluorometry: influence of parameter choice on system performance , 2000, Journal of microscopy.

[38]  W. Boron.  Intracellular pH regulation in epithelial cells. , 1986, Annual review of physiology.

[39]  I. Madshus,et al.  Regulation of intracellular pH in eukaryotic cells. , 1988, The Biochemical journal.

[40]  E. V. van Munster,et al.  φFLIM: a new method to avoid aliasing in frequency‐domain fluorescence lifetime imaging microscopy , 2004, Journal of microscopy.

[41]  A Correlation between pH and Fluorescence Lifetime of 2',7'-Bis(2-Carboxyethyl)-5(6)-carboxyfluorescein (BCECF) In Vivo and In Vitro , 2007 .

[42]  Squire,et al.  Multiple frequency fluorescence lifetime imaging microscopy , 2000, Journal of microscopy.

[43]  E. Racker,et al.  Intracellular pH measurements in Ehrlich ascites tumor cells utilizing spectroscopic probes generated in situ. , 1979, Biochemistry.

[44]  W. Leung,et al.  Synthesis and photophysical properties of new fluorinated benzo[c]xanthene dyes as intracellular pH indicators. , 2001, Bioorganic & Medicinal Chemistry Letters.

[45]  Rafael Yuste,et al.  Fluorescence microscopy today , 2005, Nature Methods.

[46]  B. Vojnovic,et al.  Multiphoton-FLIM quantification of the EGFP-mRFP1 FRET pair for localization of membrane receptor-kinase interactions. , 2005, Biophysical journal.

[47]  J. Lakowicz,et al.  Fluorescence lifetime characterization of novel low-pH probes. , 2001, Analytical biochemistry.

[48]  E. Gratton,et al.  NHE 1 Regulates the Stratum Corneum Permeability Barrier Homeostasis MICROENVIRONMENT ACIDIFICATION ASSESSED WITH FLUORESCENCE LIFETIME IMAGING * , 2002 .

[49]  A. Bergmann,et al.  Multispectral fluorescence lifetime imaging by TCSPC , 2007, Microscopy research and technique.

[50]  P. French,et al.  Time-resolved fluorescence microscopy , 2005 .

[51]  F. Waharte,et al.  Correlated fluorescence lifetime and spectral measurements in living cells , 2007, Microscopy research and technique.

[52]  Enrico Gratton,et al.  Two-photon fluorescence lifetime imaging of the skin stratum corneum pH gradient. , 2002, Biophysical journal.

[53]  H Szmacinski,et al.  Optical measurements of pH using fluorescence lifetimes and phase-modulation fluorometry. , 1993, Analytical chemistry.

[54]  R. Tsien,et al.  Cytoplasmic pH and free Mg2+ in lymphocytes , 1982, The Journal of cell biology.