scholarly journals Towards mapping the 3D genome through high speed single-molecule tracking of functional transcription factors in single living cells

Methods ◽  
2020 ◽  
Vol 170 ◽  
pp. 82-89 ◽  
Author(s):  
Adam J.M. Wollman ◽  
Erik G. Hedlund ◽  
Sviatlana Shashkova ◽  
Mark C. Leake
Keyword(s):  
Transcription Factors ◽  
Single Molecule ◽  
High Speed ◽  
Living Cells ◽  
Single Molecule Tracking ◽  
3D Genome ◽  
Single Living Cells ◽  
Single Living

scholarly journals Mapping the 3D genome through high speed single-molecule tracking of functional transcription factors in single living cells

2019 ◽  
Author(s):  
Adam J. M. Wollman ◽  
Erik G. Hedlund ◽  
Sviatlana Shashkova ◽  
Mark C. Leake
Keyword(s):  
Single Molecule ◽  
High Speed ◽  
Fluorescent Protein ◽  
Super Resolution ◽  
Living Cells ◽  
3D Genome ◽  
Straightforward Method ◽  
Single Genome ◽  
Position Data ◽  
Single Living Cells

AbstractHow genomic DNA is organized within the cell nucleus is a long-standing question. We describe a single-molecule bioimaging method with super-resolution localization precision and very rapid millisecond temporal resolution, coupled to fully quantitative image analysis tools, to help to determine genome organization and dynamics using budding yeast Saccharomyces cerevisiae as a model eukaryotic organism. We utilize astigmatism imaging, a robust technique that enables extraction of 3D position data, on genomically encoded fluorescent protein reporters that bind to DNA. Our relatively straightforward method enables snapshot reconstructions of 3D architectures of single genome conformations directly in single functional living cells.

High-speed separation system of randomly suspended single living cells by laser trap and dielectrophoresis

2001 ◽  
Vol 22 (2) ◽  
pp. 283-288 ◽  
Author(s):  
Fumihito Arai ◽  
Akihiko Ichikawa ◽  
Masanobu Ogawa ◽  
Toshio Fukuda ◽  
Koji Horio ◽  
...  
Keyword(s):  
High Speed ◽  
Living Cells ◽  
Separation System ◽  
Laser Trap ◽  
Single Living Cells ◽  
Single Living

Single molecule detection in single living cells

2000 ◽  
Author(s):  
Tyler A. Byassee ◽  
Warren C. W. Chan ◽  
Shuming Nie
Keyword(s):  
Single Molecule ◽  
Living Cells ◽  
Single Molecule Detection ◽  
Single Living Cells ◽  
Single Living

Application of FRAP and digitized fluorescence microscopy to problems in cell membrane dynamics

1988 ◽  
Vol 46 ◽  
pp. 118-119
Author(s):  
K. Jacobson ◽  
A. Ishihara ◽  
B. Holifield ◽  
F. Zhang
Keyword(s):  
Fluorescence Microscopy ◽  
Cell Biology ◽  
Extended Period ◽  
Dynamic Structure ◽  
Living Cells ◽  
Membrane Dynamics ◽  
Molecular Cell Biology ◽  
Image Averaging ◽  
Single Living Cells ◽  
Single Living

Our laboratory is concerned with understanding the dynamic structure of the plasma membrane with particular reference to the movement of membrane constituents during cell locomotion. In addition to the standard tools of molecular cell biology, we employ both fluorescence recovery after photo- bleaching (FRAP) and digitized fluorescence microscopy (DFM) to investigate individual cells. FRAP allows the measurement of translational mobility of membrane and cytoplasmic molecules in small regions of single, living cells. DFM is really a new form of light microscopy in that the distribution of individual classes of ions, molecules, and macromolecules can be followed in single, living cells. By employing fluorescent antibodies to defined antigens or fluorescent analogs of cellular constituents as well as ultrasensitive, electronic image detectors and video image averaging to improve signal to noise, fluorescent images of living cells can be acquired over an extended period without significant fading and loss of cell viability.

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