Articles NUI Galway Flow Cytometry Core Facility
The NUIG Flow Cytometry Core Facility is proud to announce the delivery and successful installation of its newest acquisition, the ImageStream®X Mark II Imaging Flow Cytometer, funded by an SFI Infrastructure award.
The revolutionary Amnis ImageStream®X Mark II Imaging Flow Cytometer (Merck Millipore) combines the speed, sensitivity, and phenotyping abilities of flow cytometry with the detailed imagery and functional insights of microscopy. This unique combination provides the tools for numerous applications that cannot be pursued using either technique alone.
Flow cytometry is a technology that allows quantitative and qualitative analysis of cell populations at a single cell level, providing multi-parametric data based on measurements of scattered light and fluorescent signals produced by cells as they pass through a laser light source. The addition of the image acquisition feature of Imaging Flow Cytometry greatly increases the amount and value of the information obtained from each experiment when compared to either of the techniques alone (Flow Cytometry and Microscopy). This instrument produces multiple high-resolution images of every cell as it flows (at a rate of up to 5000 cells per second), including brightfield and darkfield (SSC), and up to 10 fluorescent markers with sensitivity exceeding conventional flow cytometers. When compared to conventional Flow Cytometry, researchers can obtain numerous additional parameters on their samples and quantify the intensity, specific location, and distribution of signals within tens of thousands of cells per sample. These include cell morphology, nuclear shape and subcellular localization and distribution of target molecules which enable for multiple features to be analysed in great detail. The majority of the dyes used in conventional Flow Cytometry can also be detected by the instrument with compensation being also applied to the images, rendering the users with the ability to use many more dyes than they could ever use when performing microscopic analysis. Furthermore, it allows for the visual confirmation that rare events are real cells and not just artefacts.
Our system has 4 lasers (405nm, 488nm, 561nm, 642nm and 785nm (SSC)), three different objectives (20x, 40x and 60x) and the extended depth of field option (EDF). EDF keeps the depth of cell in focus without loss of fluorescence sensitivity and can be of great value when imaging FISH spots or other sub-cellular features. The data analysis software, IDEAS®, offers powerful tools for graphical representation and quantification of more than 85 parameters per cell. Coupled with the short acquisition time, this technology allows the analysis of statistically relevant numbers of images, which is not feasible using classical microscopy.
Taken together, the capabilities of the ImageStream®X Mark II system make it equivalent or superior to traditional flow applications for multi-parameter cell/particle analysis while also integrating the scope of flow cytometry and microscopy.
Typical applications include the study of:
- Protein internalization, nuclear translocation and subcellular localization
- DNA damage response
- Nuclear architecture
- Cell signalling
- Cell-cell interactions and immune synapse formation
- Cell cycle
- Nanoparticle uptake
We are keen to talk with researchers in NUI Galway and their collaborators at other institutions about potential applications of imaging flow cytometry to their ongoing and planned projects.
For more information on the capabilities of this equipment and consultation on how it can be of value to your research programme, please contact Dr. Joana Cabral at firstname.lastname@example.org