HCA Modules

Translocation Assays

We have developed automated solutions for the translocation assays commonly used in target identification/validation and compound screening in drug discovery.

  • Nucleus-Cytoplasm translocation
  • Cytoplasm-Vesicle translocation

We have research collaborations in which we have custom developed solutions for Nucleus-Cytoplasm translocation for more difficult primary cells (found below).

Commercial Partners

Our conventional translocation analysis code has been integrated into the HCA cellular imaging systems of:

  • Molecular Devices (formerly Axon Instruments) (http://www.axon.com) are a major biotechnology instrumentation company that CSIRO has worked with in the development of their ImageXpressTM cellular screening system.

If you would like to licence this module for integration into your software platform please see the contact details below.

Custom Solutions

We develop custom solutions for non-standard assays. If you’d like a quote please see the contact details below.

Nucleus-Cytoplasm translocation for commonly used cell lines

A typical cytoplasm channel image with densely packed cells from a common cell line. When well separated, these cells are roughly elliptical in shape and the cytoplasm is symmetrically located around the nucleus.

 

In widefield microscopy, it is difficult to segment the cytoplasm directly. So cytoplasm measurements are usually taken using a surrogate for the cytoplasm – an isotropic “donut” region around the nucleus.

 

When cells are densely packed, isotropic donuts will include some background. Our solution allows the use of anisotropic “donuts”. This reduces the likelihood that low signal background will bias  the cytoplasm measurement.

 

fluore1  fluore2  fluore3

Nucleus-Cytoplasm translocation for more difficult primary cells

A typical cytoplasm channel image with densely packed primary cells. Even when well separated, these cells are not elliptical in shape and nor is the cytoplasm symmetrically located around the nucleus.

 

 We have developed a new cytoplasm surrogate using adaptive donuts. These expand within the cytoplasm differentially to avoid the background. This significantly reduces the false alarm rate from incorrect cytoplasm measurements.
 C_-_10_fld_3_cytoplasm_sub C_-_10_fld_3_adaptive_cytoplasm_overlay_sub

Cytoplasm-vesicle translocation

3_Cyto_Stain_sub  3_Cyto_Stain_dot_binary_sub  3_Cyto_Stain_dot_overlay_sub
A typical cytoplasm-to-vesicle translocation assay – a negative result is shown above, a positive result is shown below.  Standard solutions just count the vesicles/dots in the full image. So heterogeneous responses cannot be detected.   Our solution allocates each vesicle/dot to a single cell. So atypical cells with high or low response can be detected.
4_Cyto_Stain_sub  4_Cyto_Stain_dot_binary_sub  4_Cyto_Stain_dot_overlay_sub