Cellgraph - Visualizing Dynamic Gene Expression Networks with Luciferase Markers
Living cell activities are regulated by the complex gene expression
networks, which have been analyzed and illuminated with varieties of
reporter gene methods using fluorescent proteins (GFP etc.) and/or
bioluminescent proteins (firefly luciferase etc.).
Luciferases offer some advantages over fluorescent proteins, i.e. minimizing phototoxic effects causing damage to living cells, higher S/N without undesirable autofluorescence background emission or scatter generated by intracellular components.
Luciferase markers are also advantageous in long-term (up to several days) assay of living cells due to relatively low cytotoxicity of the substrate luciferin.
Cellgraph AB-3000B is an imaging system designed to detect extremely weak bioluminescent signals from a single cell. In combination with luciferase markers, Cellgraph can visualize the cell dynamics - time dependent changes of transcriptional activities in individual cells and localization to inner cell components (organelles).
Advanced Features of Cellgraph
Cellgraph provides a variety of advanced features to serve living
- The ultrasensitive cooled and back-illuminated EMCCD (Electron Multiplying CCD) offers single photon sensitivity, certified with a patented procedure to measure total luminescence intensity as photon counts.
- The incubation system enables long-term culture under controlled temperature and 5% carbon dioxide gas circulation.
- The multicolor detection system is equipped for multi reporter assay with up to 2 different color filters, which enhances sensitivity to extra weak bioluminescence and gives an internal standard basis.
- Versatile image capturing modes include; time-lapse capture at user-set interval, z-stepping mode to optimize Z-stage position (focal plane), switching mode to capture in a dark view and an illuminated view (a luminescent image and a bright field image) alternately.
- Powerful tools for data analysis, editing, and presentation are also provided, i.e. grid measurement at view areas divided into the lattice of user-specified size, spot (ROI) measurement at user-selected areas of a chosen shape (ellipse, rectangle, polygon, or spline curve) and sizes, a movie maker converting time-lapse images into a movie file of AVI format.
Applications of Cellgraph AB-3000B
- Cellgraph has a huge potential to serve various fields of
researches focussing gene expression in living cells, tissues,
i.e. studies on chronobiology, development, cell differentiation, drug design, cancer, etc.
- An original paper by Hoshino, H. et al, which refers to Cellgraph, has been published on Nature Methods in August.
also provides Kronos Dio, a luminometer for real-time reporter assay;
- Kronos Dio is a Luminometer for real-time reporter assay with 8 dish incubator and multicolor measurement system. Kronos Dio provides a long-term multi culture monitoring tool which can be used in screening, examining different culture conditions on gene expression, etc.
More information: KronosDio.pdf
More information: Cellgraph.pdf
More information in general: