A high-parameter spectral cytometer, the ID7000 delivers comprehensive information about heterogeneous cell populations, with high sensitivity to detect dim and rare populations.
Expanding the boundaries of cell analysis, the ID7000 can be configured with up to 7 lasers and 188 detectors, the most of any flow cytometer available today. The system enables researchers to perform experiments using 44 colors, limited only by the fluorochromes available. The choice of lasers include blue (488 nm), red (637 nm), violet (405 nm), yellow-green (561 nm), ultraviolet (355 nm), deep ultraviolet (320 nm), and infrared (808 nm).
The ID7000 provides capabilities to simplify high-parameter experiments and streamline multicolor workflows. Novel features include the Spectral Reference Library and the Autofluorescence Finder. The Spectral Reference Library stores information from single-positive controls for each reagent for future use, so researchers need to run controls only once, saving time. With the Auto-fluorescence Finder, researchers can identify and subtract contributions from multiple autofluorescent populations for higher fidelity data and more accurate visualization.
Spectral Reference Library
Software wizards and tools simplify instrument controls from startup to shutdown, automate quality control, and optimize experiment creation. For example, the system’s AutoSampler includes a unique, active agitation capability to keep cells in suspension during acquisition. Also, an active cooling function further reduces variability and prevents sample degradation over time. Software and automation manage error conditions such as bubbles, clogs, and low sample volume, and enable walkaway operation.
The AutoSampler supports a wide variety of standard and deep 96- and 384-well plates, and up to twenty-four (12 x 75-mm) 5-mL tubes in the tube loader.
In the above graph, the blue line shows the event rate when the active agitation is applied for 2 seconds at 180-second intervals, keeping the sample in suspension. The brown line shows agitation for 2 seconds only, resulting in cells settling over time.
Harness Complex Data
The ID7000 system builds on Sony’s experience with spectral analysis and simplifies many operations, even for complex experiments. The system collects the entire spectrum of light from 360 nm to 920 nm, which allows researchers to separate fluorochromes into pure signals to more accurately measure data. In comparison, conventional flow cytometry uses color compensation to determine each fluorescence intensity by subtracting overlapping fluorescence, resulting in gaps in the collected light. Thus, some of the data is lost.
Optics design and spectral technology make it possible for researchers to use both spectrally adjacent fluorochromes and fluorescent proteins in a panel—a capability that is not practical using a traditional cytometer.
A powerful capability of spectral technology is unmixing. Spectral unmixing objectively manages overlap between fluorescence spectra to improve data accuracy. A novel auto-unmixing capability, and new algorithms, speed processing for ease of operation. Spectral unmixing eliminates the subjectivity and complexity associated with manual compensation methods.
Spectral unmixing separates each spectral fingerprint for complete and optimal visualization of fluorochromes