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Quantifying E. coli Proteome and Transcriptome with Single-Molecule Sensitivity in Single Cells
System-wide analyses of protein and mRNA expression in individual cells with single-molecule sensitivity using a newly constructed yellow fluorescent protein fusion library for Escherichia coli found that almost all protein number distributions can be described by the gamma distribution with two fitting parameters which, at low expression levels, have clear physical interpretations as the transcription rate and protein burst size. Expand
Probing Transcription Factor Dynamics at the Single-Molecule Level in a Living Cell
In a living Escherichia coli cell, specific binding of a lac repressor, labeled with a fluorescent protein, to a chromosomal lac operator is observed and the kinetics of binding and dissociation of the repressor in response to metabolic signals are measured using single-molecule detection techniques. Expand
Genome-Wide Detection of Single-Nucleotide and Copy-Number Variations of a Single Human Cell
A new amplification method—multiple annealing and looping-based amplification cycles (MALBAC)—that offers high uniformity across the genome that achieves 93% genome coverage ≥1x for a single human cell at 25x mean sequencing depth is reported. Expand
Probing Gene Expression in Live Cells, One Protein Molecule at a Time
It is found that the protein molecules are produced in bursts, with each burst originating from a stochastically transcribed single messenger RNA molecule, and that protein copy numbers in the bursts follow a geometric distribution. Expand
Label-Free Biomedical Imaging with High Sensitivity by Stimulated Raman Scattering Microscopy
Three-dimensional multiphoton vibrational imaging technique based on stimulated Raman scattering (SRS) is reported, which offers background-free and readily interpretable chemical contrast in biomedical imaging and shows a variety of biomedical applications. Expand
Linking stochastic dynamics to population distribution: an analytical framework of gene expression.
This model allows for the extraction of kinetic parameters of gene expression from steady-state distributions of protein concentration in a cell population, which are available from single cell data obtained by flow cytometry or fluorescence microscopy. Expand
Three-Dimensional Vibrational Imaging by Coherent Anti-Stokes Raman Scattering
Confocal [1] and multiphoton [2] fluorescence microscopy have become powerful techniques for threedimensional imaging of chemical and biological samples, especially for live cells. This coincidesExpand
Single Molecule Imaging of Transcription Factor Binding to DNA in Live Mammalian Cells
Reflected light-sheet microscopy (RLSM), a fluorescence microscopy method allowing selective plane illumination throughout the nuclei of living mammalian cells, is developed and single-molecule measurements and statistical analysis revealed dynamic properties of transcription factors. Expand
Potent Neutralizing Antibodies against SARS-CoV-2 Identified by High-Throughput Single-Cell Sequencing of Convalescent Patients’ B Cells
It is shown that human neutralizing antibodies could be efficiently discovered by high-throughput single B-cell sequencing in response to pandemic infectious diseases. Expand
Stochastic protein expression in individual cells at the single molecule level
A microfluidic-based assay is demonstrated that allows real-time observation of the expression of β-galactosidase in living Escherichia coli cells with single molecule sensitivity and shows that protein production occurs in bursts, with the number of molecules per burst following an exponential distribution. Expand