Xcyto 10-workshop

On 12 September, a workshop and introduction lecture on the Xcyto 10 will be held. The system is installed at CFIM for anybody to test. 

If you have any questions, please contact Lars D. Johansen at LAJ@chemometec.com.

the introduction lecture will take place in Faculty Club (16.6) at 10:30. 

Xcyto® 10

Quantitative Cell Imager
Made in Denmark

The Danish company ChemoMetec A/S has developed a new automated microscope that is available in a free trial period at CFIM (Panum Institute). You are invited to attend the seminar 10:30 September 12th at ? outlining how to use the Xcyto® 10 system. The Xcyto® 10 will be available at CFIM at least two months and hands on training is offered.

The Xcyto® 10 image cytometer combines the statistical power and fluorescence sensitivity of standard flow cytometry with the spatial resolution and quantitative morphology of digital microscopy. This unique combination allows for new applications that would be unfeasible using the two methods separately. The Xcyto® 10 system integrates image acquisition, image analysis, feature extraction and data presentation in one step, providing seamless quantitative data of individual cells. The system uses relatively low magnification (4x and 20x) enabling acquisition of data from a large amount of cells to use in statistical analysis of results.

Cytometry

  • Statistical robustness
  • Flexibility in multicolor panel choice
  • High sensitivity for detection of dim photometric features
  • Segregation of cell populations and organelles in an intuitive and automated interface

Quantitative Microscopy

  • Spatial localization and abundance of fluorescence
  • Translocation and colocalization of markers
  • Cellular or organellar morphometric and physiometric features
  • Quantifiable insights into dynamic processes in living cells

How Xcyto® 10 resolves quantitative data of individual cells

Cytometric features from each cell are automatically extracted from the images and presented in plots.

Suspension cells:

This method relies on the UV phase channel to segment the cells in suspension.

  • After standard staining your cells with fluorescent probes load a Xcyto® 2-Chamber slide.
  • Select to use the 4x or 20x objective and the channels to acquire.
  • Insert slide and press run.
  • After approximately 1 minute the first data will be presented as images and plots.

Adherent cells:

This method relies on the DAPI channel to find the nucleus of each cell an on the BlueMask™ channel to find the cell borders.

  • Seed your cells on 4 and 8 well culture slides or on coverslip glasses overnight.
  • Fix the cells and stain with fluorescent probes.
  • After wash with PBS stain the cells with BlueMask™ and DAPI for 30 minutes.
  • Wash with PBS and mount coverslips on either culture slides or Xcyto® 2-Sample Slides.
  • Select to use the 4x or 20x objective and the channels to acquire.
  • Insert slide and press run.
  • After approximately 2 minutes the first data will be presented as images and plots.

Selected examples of applications

  • Endocytosis and Autophagy: Tracking e.g. phagocytosis on markers for autophagy.
  • Biochemistry: Trace and quantify enzymatic activity.
  • Cell Cycle and Proliferation: Multidimensional analysis and spatial resolution of probes.
  • Parasitology and Virology: E.g. detection of infected cells.
  • Cell Signaling: E.g. nuclear translocation of transcription factor.
  • Protein-Protein Interaction: E.g. FRET, BiFC.
  • Multiplexing: E.g. Fluorescence In Situ Hybridization (FISH) coupled with immunofluorescence.
  • DNA Damage and Repair: E.g. spot abundance of ɣH2AX recruitment to DNA lesions.
  • Immunology: Multicolor cytotoxicity and immunophenotyping coupled with imagery.
  • Morphology: Changes in cell shape in response to e.g. macrophage activation and response to drugs.
  • Protein Expression and Localization: Quantification of subcellular protein localization in single cells.
  • Cell Death: E.g. apoptosis and necrosis markers in intact (non-trypsinized) cells.

36+ Channels

Maximum 7 fluorescence channels at a time