Main ContentAmnis ImageStreamX MkII
The Amnis ImageStreamX is equipped with two CCD cameras, which can acquire up to 12 images simultaneously of each cell or particle. However, brightfield images must be collected on both Camera 1 and Camera 2 to precisely align all 12 images during analyses. Therefore, the system can locate and quantify the intensity and distribution of up to 10 different fluorescent signals on, within, or between cells in cell conjugates. Applications include:
- Intracellular co-localization
- Nuclear translocation
- Internalization and phagocytosis
- Cell-cell interaction
- Cell death and autophagy
- Cell cycle and mitosis
For more information, visit the manufacturer's website: https://cytekbio.com/pages/imagestream
When planning an imaging flow cytometry experiment and choosing fluorochromes, it is important to know the fluorochromes' specific excitation and emission wavelengths, as well as the lasers, filters, and detectors available on the instrument. Below is the information for the Amnis ImageStreamX MkII.
| Amnis ImageStreamX MkII Configuration and Fluorochromes |
| Camera | Excitation Laser | Channel | Bandpass Filter | Ideal dyes | Additional Fluorochromes |
| Camera 1 | Bright field | Ch1 | | Bright Field | Bright Field |
Blue 488 nm | Ch2 | 528/65(480-560) | FITC, AF488, GFP | CFSE, YFP, DyLight488, PKH67-membrane label, LysoTrackerGreen |
| Ch3 | 577/35560-595 | PE | Cy3, DSRed, PKH67-membrane label SYTOX Orange |
| Ch4 | 610/30595-642 | PE-TexasRed* | RFP, PE-AF610, eFluor625, Qdot625* Ch4 & Ch10 BF** |
| Ch5 | 702/85642-745 | PE-Cy5*, PE-AF647 | PI, 7AAD, PerCP-Cy5.5, eFluor 650, Draq 5 |
| Ch6 | 765/40745-785 | PE-Cy7*, PE-AF750 | or use for side scatter (SSC) |
| Camera 2 | Violet 405 nm | Ch7 | 457/45435-505 | DAPI, AF405, BV421 | Hoechst, eFluor 450, DyLight 405, CFP, Live/Dead violet |
| Ch8 | 537/65505-570 | BV510 | AF430, eFluor506, Qdot 525, Cascade yellow |
| Ch9 | 582/25570-595 | Bright Field | Qdot565, Qdot585, eFluor565 |
| Ch10 | 610/30595-642 | | Qdot 605, Qdot625, eFluor625, BV605 Ch4 & Ch10 BF** |
| Ch11 | 702/85642-745 | | BV711, eFluor700, Qdot 705* |
| Ch12 | 765/40745-785 | | Qdot 800*, BV786 or use for side scatter |
Red 640 nm | Ch11 | 702/85642-745 | AF647, AF660, AF680 | APC, Cy5, AF660, Draq 5*, DyLight649, DyLight680, PE-AF647*, PE-Cy5 |
| Ch12 | 765/40745-785 | APC-Cy7, APC-AF750 | APC-H7, Cy7, APC-eFluor780, AF-750, DyLight750, PE-Cy7* PE- AF750* or use for side scatter |
*It is important to remember that many fluorescent dyes are excited by more than one laser and fluoresce in multiple channels. It is very helpful to use a spectral viewer when planning an experiment.
** There are options to switch the Brightfield to different channels if needed (Ch1 & Ch9, Ch2 & Ch8, Ch4 & Ch10, and Ch6 & Ch12).
Ideal fluorochromes for this instrument are AF488, PE, PE-TexasRed, SSC-Ch6, DAPI (or BV421), AF647, and APC-Cy7.