Quanterix Akoya PhenoImager HT 2.0

Quanterix Akoya PhenoImager HT 2.0

The Quanterix Akoya PhenoImager HT 2 is a whole-slide imaging system built for quantitative spatial phenotyping and high-throughput tissue analysis.

Core System Specifications

• Optical Magnification: Supports digital whole-slide scanning at 10x (1.0 μm/pixel), 20x (0.5 μm/pixel), and 40x (0.25 μm/pixel) resolutions.
• Slide Capacity and Automation: Features a touchless, automated loader that accommodates up to 80 slides simultaneously. The system utilizes continuous loading technology to permit slide replacement during active scanning operations.
• Imaging Modalities (Brightfield, MSI or Motif whole slide scans): Acquires images in either brightfield or fluorescence formats from tissue sections or tissue microarrays (TMAs).  For fluorescence, the system uses either long-pass filters in conjunction with a liquid crystal tunable filter (LCTF) or a set of narrow band filters to collect multispectral images.
• Software Integration: Links to inForm and phenoptr semi-automated tissue analysis packages, which use machine-learning algorithms to execute tissue segmentation, cell segmentation, and phenotypic scoring.  Whole slide unmixed multispectral images can be created and analyzed with Indica Labs Halo.
• File Output: Exports data in the standardized Akoya whole-slide multi-resolution tiled format (.QPTIFF) or individual multispectral field formats (.im3).

7 Color Multispectral Imaging and Onboard Unmixing

The platform integrates reference library spectral profiles directly into the scanning sequence:

• Protocol Loading: The generated Spectral Library is uploaded into the instrument software prior to beginning a scanning batch.
• Multispectral Acquisition: The LCTF system records raw multiplexed emission data across multiple discrete wavelength bands during the tissue scan.
• Linear Matrix Solving: The system software applies a linear unmixing algorithm on a pixel-by-pixel basis. The total measured light intensity at each pixel is calculated as a linear combination of the individual dye components from the library.
• Real-Time Processing: The instrument processes these matrix equations dynamically alongside real-time image stitching as data capture proceeds across the slide.

9 Color Multispectral Imaging Capabilities

The PhenoImager HT integrates a hardware-based Liquid Crystal Tunable Filter (LCTF) in its emission optical path to manage spectral overlap in multiplexed panels.

• Wavelength Spectrum: Operates across a multispectral range of 440 nm to 780 nm, capturing emission data across more than 40 discrete wavelength bands.
• Multiplex Capacity: Isolates up to 9 distinct colors (typically DAPI plus 6 to 8 fluorescent biomarkers, such as Opal dye sets) within a single tissue section.
• Spectral Unmixing: Leverages established reference libraries (derived from single-stained and unstained tissues) to calculate and separate overlapping emission signatures. The instrument software processes this math on a pixel-by-pixel basis.
• Autofluorescence Isolation: Treats tissue-specific background autofluorescence as an independent channel during the unmixing calculations. This separates the background noise from target fluorophore signals to increase the signal-to-noise ratio.