Non-invasive membrane potential measurements using endogenous markers

Author(s):  
Georgi I. Petrov ◽  
Vladislav V. Yakovlev
1996 ◽  
Vol 150 (3) ◽  
pp. 445-456 ◽  
Author(s):  
M Mattioli ◽  
B Barboni ◽  
L Gioia

Abstract We have studied the changes in membrane potential induced by LH in cumulus and granulosa cells isolated from sheep antral follicles. The investigation was carried out by using a non-invasive technique based on the use of a membrane potential sensitive probe, bis-oxonol. The membrane potential of mural granulosa cells was totally unaffected by LH, while that of cumulus or corona cells showed a marked depolarisation, starting 2–3 min after the addition of the hormone and plateauing after 5–10 min. None of the cells tested reacted to FSH. In the second part of the experiment the role of protein kinase A (PKA) and protein kinase C (PKC) in mediating the effect of LH was studied. The selective activation of PKA or PKC induced in cumulus-corona cells a rapid hyperpolarisation due to increased Cl and K conductance respectively. By contrast, the simultaneous activation of the two kinases induced a rapid membrane depolarisation due to the progressive decrease in K conductance. The activation of each kinase or their combined stimulation did not induce any change in the membrane potential of mural granulosa cells. These data demonstrated that LH has a depolarising effect regionally circumscribed to cumulus-corona cells and that this depolarisation depends on a reduction of K conductance caused by the activation of PKA and PKC. Journal of Endocrinology (1996) 150, 445–456


2019 ◽  
Author(s):  
Ella Matamala ◽  
Cristian Castillo ◽  
Juan Pablo Vivar ◽  
Sebastian Brauchi

AbstractEukaryotic cells are complex systems compartmentalized in membrane-bound organelles. Visualization of organellar electrical activity in living cells requires both a suitable reporter and non-invasive imaging at high spatiotemporal resolution. Here we present hVoSorg, an optical method to monitor changes in the membrane potential of subcellular membranes. This method takes advantage of a FRET pair consisting of a membrane-bound voltage-insensitive fluorescent donor and a colorless voltage-dependent acceptor that rapidly moves across the membrane in response to changes in polarity. Compared to the currently available techniques, hVoSorg has advantages including simple and precise subcellular targeting, the ability to record from individual organelles, and the potential for optical multiplexing.SummaryBy adapting a hybrid-FRET voltage sensor we report here the resting membrane potential of different organelles in living cells.


2019 ◽  
Vol 2 (1) ◽  
pp. 93-97 ◽  
Author(s):  
Eugen Gheorghiu

Abstract Non-invasive, label-free assessment of membrane potential of living cells is still a challenging task. The theory linking membrane potential to the low frequency α dispersion exhibited by suspensions of spherical shelled particles (presenting a net charge distribution on the inner side of the shell) has been pioneered in our previous studies with emphasis on the permittivity spectra. Whereas α dispersion is related to a rather large variation exhibited by the permittivity spectrum, we report that the related decrement presented by the impedance magnitude spectrum is either extremely small, or occurs (for large cells) at very small frequencies (~mHz) explaining the lack of experimental bioimpedance data on the matter. We stress that appropriate choice of the parameters (as revealed by the microscopic model) may enable access to membrane potential as well as to other relevant parameters when investigating living cells and charged lipid vesicles. We analyse the effect on the low frequency of the permittivity and impedance spectra of: I. Parameters pertaining to cell membrane i.e. (i) membrane potential (through the amount of the net charge on the inner side of the membrane), (ii) size of the cells/vesicles, (iii) conductivity of the membrane; II. Parameters of the extra cellular medium (viscosity and conductivity). The applicability of the study has far reaching implications for basic (life) sciences (providing non-invasive access to the dynamics of relevant cell parameters) as well as for biosensing applications, e.g. assessment of cytotoxicity of a wide range of stimuli.


Author(s):  
H.W. Deckman ◽  
B.F. Flannery ◽  
J.H. Dunsmuir ◽  
K.D' Amico

We have developed a new X-ray microscope which produces complete three dimensional images of samples. The microscope operates by performing X-ray tomography with unprecedented resolution. Tomography is a non-invasive imaging technique that creates maps of the internal structure of samples from measurement of the attenuation of penetrating radiation. As conventionally practiced in medical Computed Tomography (CT), radiologists produce maps of bone and tissue structure in several planar sections that reveal features with 1mm resolution and 1% contrast. Microtomography extends the capability of CT in several ways. First, the resolution which approaches one micron, is one thousand times higher than that of the medical CT. Second, our approach acquires and analyses the data in a panoramic imaging format that directly produces three-dimensional maps in a series of contiguous stacked planes. Typical maps available today consist of three hundred planar sections each containing 512x512 pixels. Finally, and perhaps of most import scientifically, microtomography using a synchrotron X-ray source, allows us to generate maps of individual element.


Author(s):  
R H. Selinfreund ◽  
A. H. Cornell-Bell

Cellular electrophysiological properties are normally monitored by standard patch clamp techniques . The combination of membrane potential dyes with time-lapse laser confocal microscopy provides a more direct, least destructive rapid method for monitoring changes in neuronal electrical activity. Using membrane potential dyes we found that spontaneous action potential firing can be detected using time-lapse confocal microscopy. Initially, patch clamp recording techniques were used to verify spontaneous electrical activity in GH4\C1 pituitary cells. It was found that serum depleted cells had reduced spontaneous electrical activity. Brief exposure to the serum derived growth factor, IGF-1, reconstituted electrical activity. We have examined the possibility of developing a rapid fluorescent assay to measure neuronal activity using membrane potential dyes. This neuronal regeneration assay has been adapted to run on a confocal microscope. Quantitative fluorescence is then used to measure a compounds ability to regenerate neuronal firing.The membrane potential dye di-8-ANEPPS was selected for these experiments. Di-8- ANEPPS is internalized slowly, has a high signal to noise ratio (40:1), has a linear fluorescent response to change in voltage.


Author(s):  
Leslie M. Loew

A major application of potentiometric dyes has been the multisite optical recording of electrical activity in excitable systems. After being championed by L.B. Cohen and his colleagues for the past 20 years, the impact of this technology is rapidly being felt and is spreading to an increasing number of neuroscience laboratories. A second class of experiments involves using dyes to image membrane potential distributions in single cells by digital imaging microscopy - a major focus of this lab. These studies usually do not require the temporal resolution of multisite optical recording, being primarily focussed on slow cell biological processes, and therefore can achieve much higher spatial resolution. We have developed 2 methods for quantitative imaging of membrane potential. One method uses dual wavelength imaging of membrane-staining dyes and the other uses quantitative 3D imaging of a fluorescent lipophilic cation; the dyes used in each case were synthesized for this purpose in this laboratory.


2001 ◽  
Vol 120 (5) ◽  
pp. A266-A266
Author(s):  
R BUTLER ◽  
B ZACHARAKIS ◽  
D MOORE ◽  
K CRAWFORD ◽  
G DAVIDSON ◽  
...  

2001 ◽  
Vol 120 (5) ◽  
pp. A491-A491 ◽  
Author(s):  
A LEODOLTER ◽  
D VAIRA ◽  
F BAZZOLL ◽  
A HIRSCHL ◽  
F MEGRAUD ◽  
...  
Keyword(s):  

Sign in / Sign up

Export Citation Format

Share Document