Peeking Into Live Cells

Arnaud, Celia

doi:10.1021/cen-v085n023.p013

Cited by 1 publication

(5 citation statements)

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“…Molecular analysis in terms of single molecules is becoming a necessity in new tools for experimental and molecular medicine, medical genomics and proteomics, biochemistry, molecular biology and immunology [1]. The achieved detection sensitivity in analytical chemistry allows real-time measurements of a single molecule in solution or membrane or within a live cell [1][2][3][4][5][6][7].…”

Section: Discussionmentioning

confidence: 99%

“…The achieved detection sensitivity in analytical chemistry allows real-time measurements of a single molecule in solution or membrane or within a live cell [1][2][3][4][5][6][7]. Modern diagnostic assays are generally capable of detecting specific molecules below ten.…”

Section: Discussionmentioning

confidence: 99%

“…Prominent among these are a variety of technologies that rely on fluorescence in response to monochromatic light [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17].…”

Section: Introductionmentioning

confidence: 99%

“…Singlemolecule live-cell imaging is still in its early days, and opportunities are plentiful to keep many scientists busy for a long time. Originally, single-molecule work was restricted to a handful of researchers, but over time, it will become easier, because one will eventually have better probes and pushbutton devices that will make things simple enough for "everybody" to use it [1].…”

Section: Introductionmentioning

confidence: 99%

“…Quantitative understanding of molecular interactions at the level of single molecules within single cells is the next step in basic and applied biomedical research for the analysis of the dynamics and localization of molecules in a variety of physiological and pathophysiological processes [1][2][3][4][5][6][7]. However, what is the measurement time in which one is able to study just one single molecule in solution without immobilization or hydrodynamic focusing?…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Fluorescence Fluctuation Spectroscopic Approaches to the Study of a Single Molecule Diffusing in Solution and a Live Cell without Systemic Drift or Convection: A Theoretical Study

Földes-Papp

2007

CPB

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Reentries of a single molecule in the confocal, femtoliter-sized probe region (about 10(-16) L and less) are significant because during measurement times they give rise to fluctuation phenomena such as molecule number fluctuations at the single-molecule level in solution without immobilization or hydrodynamic focusing. These fluctuations are the fundamental physical process on which, for example, fluorescence correlation spectroscopy and two-color fluorescence cross-correlation spectroscopy are based. The reentries of just one molecule in the confocal probe region are theoretically examined in this original article using a hidden, continuous-time Markov model. The system is not set up to have systemic drift or convection. It is found that the reentries obey certain conditions and analytical expressions for the reentry probabilities are obtained first. In particular, the time constant of the mean value and the variance of the reentry probabilities are obtained. The fractions of non-meaningful reentries and meaningful reentries are found for these experimental situations. Therewith, the concentration dependence of the meaningful time that one can study bimolecular reactions of the selfsame molecule in the confocal probe region is derived for the first time. The meaningful time in the probe volume is proportional to the diffusion time of the selfsame molecule and related inversely to the size of the given confocal probe volume. For small molecules, i.e. small diffusion times at a given size of the confocal probe region, one needs lower concentrations of molecules of the same kind in the bulk phase, whereas large molecules can be studied at higher concentrations. The selfsame molecule scenario is compared with the molecular scenario that a second molecule enters the probe volume at random as a function of the meaningful time. The analytical solutions of the physical reentry model (mechanism) hold for the one-, two- (membrane), or three- (solution, live cell) dimensional Brownian motion.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…Prominent among these are a variety of technologies that rely on fluorescence in response to monochromatic light [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17].…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%