Activity driven fluctuations in living cells

We propose a model for the dynamics of a probe embedded in a living cell, where both thermal fluctuations and nonequilibrium activity coexist. The model is based on a confining harmonic potential describing the elastic cytoskeletal matrix, which undergoes random active hops as a result of the nonequilibrium rearrangements within the cell. We describe the probe's statistics and we bring forth quantities affected by the nonequilibrium activity. We find an excellent agreement between the predictions of our model and experimental results for tracers inside living cells. Finally, we exploit our model to arrive at quantitative predictions for the parameters characterizing nonequilibrium activity, such as the typical time scale of the activity and the amplitude of the active fluctuations.Comment: 6 pages, 4 figure

Energetics of active fluctuations in living cells

The nonequilibrium activity taking place in a living cell can be monitored with a tracer embedded in the medium. While microrheology experiments based on optical manipulation of such probes have become increasingly standard, we put forward a number of experiments with alternative protocols that, we claim, will provide new insight into the energetics of active fluctuations. These are based on either performing thermodynamic--like cycles in control-parameter space, or on determining response to external perturbations of the confining trap beyond simple translation. We illustrate our proposals on an active itinerant Brownian oscillator modeling the dynamics of a probe embedded in a living medium

Containment and reciprocity in biological systems : a putative psychophysical organising principle

The stuff of life, the living substance that is common to all biological organisms, is the aqueous society of biochemical activity ongoing in every cell in every living body. The basic biochemical ‘reactions’ of life are largely similar with variations of a theme played out in different cells living in different environment, e.g. the core biochemical metabolic processes of all life likely stem from an ancient, early-earth ancestor (Smith & Morowitz, 2004). However, even more common to life than shared biochemistry are the basic structural properties of all cells and all living organisms into complexes of compartmentalised units. In this paper, I will argue there are common feelings driving the generation of these ubiquitous structures in nature and that these feelings may constitute one of several primary forms of feeling in living systems

Force measurements with optical tweezers inside living cells

The force exerted by optical tweezers can be measured by tracking the momentum changes of the trapping beam, a method which is more general and powerful than traditional calibration techniques as it is based on first principles, but which has not been brought to its full potential yet, probably due to practical difficulties when combined with high-NA optical traps, such as the necessity to capture a large fraction of the scattered light. We show that it is possible to measure forces on arbitrary biological objects inside cells without an in situ calibration, using this approach. The instrument can be calibrated by measuring three scaling parameters that are exclusively determined by the design of the system, thus obtaining a conversion factor from volts to piconewtons that is theoretically independent of the physical properties of the sample and its environment. We prove that this factor keeps valid inside cells as it shows good agreement with other calibration methods developed in recent years for viscoelastic media. Finally, we apply the method to measuring the stall forces of kinesin and dynein in living A549 cells.Publisher PD

Wide-Field Multi-Parameter FLIM: Long-Term Minimal Invasive Observation of Proteins in Living Cells.

Time-domain Fluorescence Lifetime Imaging Microscopy (FLIM) is a remarkable tool to monitor the dynamics of fluorophore-tagged protein domains inside living cells. We propose a Wide-Field Multi-Parameter FLIM method (WFMP-FLIM) aimed to monitor continuously living cells under minimum light intensity at a given illumination energy dose. A powerful data analysis technique applied to the WFMP-FLIM data sets allows to optimize the estimation accuracy of physical parameters at very low fluorescence signal levels approaching the lower bound theoretical limit. We demonstrate the efficiency of WFMP-FLIM by presenting two independent and relevant long-term experiments in cell biology: 1) FRET analysis of simultaneously recorded donor and acceptor fluorescence in living HeLa cells and 2) tracking of mitochondrial transport combined with fluorescence lifetime analysis in neuronal processes