Flow cytometery is a powerful technique to perform study at single cell level within a large number of homo- or heterogeneous cell populations at high speed. Taking advantage of various combination of marker specific fluorescence dyes, specific cell type/stage can be identified based on the specific combination and intensity of dyes. Yet this powerful technique has its limitations, as it does not allow cell type/stage differentiation base on cellular localization/morphology as a result of the lack of any imaging component.
||The breakthrough bought about by Amnis ImageStream is the introduction of imaging capability to a flow cytometer (Multispectral Imaging Flow Cytometry).
This technology enables the identification of single cell based on fluorescence dyes intensity, morphology of the cell via the bright field imaging, as well as morphology / distribution of fluorescence marker(s).
This capability allows the ImageStream to differential cell populations that are normally indistinguishable on conventional flow cyctomter, thus make the ImageStream an extremely powerful instrument for the study of events such as translocation, internalization, cell cycle, pseudopod formation and virtually any morphological changes that can be detected by conventional transmitted light and/or epi-fluorescence imaging.
In conjunction with the powerful IDEAS® analytical software, the post-acquisition image analysis is greatly simplified. The IDEAS® allows you to quantify the morphological changes and display the results in formats that are similar to that of conventional 1-D and 2-D flow cytometry plot, only you can plot many more parameters such as area, aspect ratio, perimeter, max./min./mean/median/integrated intensity, translocation/co-localization index. You can also perfrom mathametical calculation with the features and plot the result with the 1-D or 2-D plot.
Since ImageStream is built upon a flow cytometer and behaves like one, most of the existing sample staining protocols used on a standard flow cytometer can be used to stain sample for ImageStream with minimal modification. However, since the ImageStream utilize a slower fluidic system to facilitate image capturing, the requirement for cell density is different. Please see the sample preparation guide for detail information.
With proper training, the ImageStream is a user operatable instrument. The operation training generally takes around two ours, and includes instrument start up, instrument setup, sample switching, instrument shut down, as well as simple troubleshooting. All the analyses, as well as spectral compensation, are performed post-acquisition using the analytical software.
** Free "Proof-of-concept" run is available upon request, please contact the core facility manager, Albert Tai, to schedule. **
- Instrument operation training: 2hrs per session
- Data analysis consultant, if applicable: same as instrument rate
Workstations for data analysis are available free of charge.
- Three solid-state excitation laser
488nm sapphire laser @ 200mW
405nm violet laser @ 350mW
658nm red laser @ 90mW
- Five fluorescence emission filters
||Darkfield / Side scatter only
|DAPI, Hoechst, Pacific Blue, Cascade Blue and Alexa Fluor 405 & 430
|FITC, Alexa fluor 488, (E)GFP, Syto Green, Spectrum Green, YFP, Sybr Green and Pacific Orange
|PE, Cy3, Alexa Fluor 546 & 555 and DsRed
|7-AAD, PE/Alexa Fluor 610, PI, PE/Texas Red and ECD
- 36x 0.75NA objective equivalent, 0.5 x 0.5μm per pixel pf CCD camera, 45μm image view width.
- Up to 100-200 events per second and 6 images per event
Please visit the manufacturer website
for additional information.