NovaISM

Intensity-weighted ISM-FLIM image. Colorscale indicates the fast lifetime contrast ranging from blue (shorter lifetimes) to red (longer lifetimes) sample: neuronal hippocampal culture, labeling of mitochondrial marker TOM20 with Cy2 (lifetime 1.3 ns) and synamptic vesicles marker SYPT1 with OregonGreen (lifetime 3.5 ns). Samples prepared by Dr. Eman Abbas, Rizzoli lab, UMG Goettingen.

In NovaISM computational sectioning is performed simultaneously with lifetime species separation allowing for determineing in –focus and out-of-focus contributions independently for each lifetime species. For further analysis then only the infocus part can be used. Upper side: SYPT1, lower side: TOM20, left side: out-of-focus, right side: in-focus.

Deconvolution is then performed only on the in-focus part. In this way we solve the problems arising from out-of-focus light when deconvolution algorithms are applied in thick samples. The end result is an image with high resolution, extremely high contrast and simultaneous marker multiplexing based on lifetimes. SYPT1 in cyan, TOM20 in yellow.

A simple schematic of the principle of computational sectioning. The distribution of light on the PDA-23 detector is fitted with 2 Gaussian of different widths. The in-focus part correspond to the narrow Gaussian and the out-of-focus part to the broader Gaussian. Parameters are estimated automatically by the software while the user can always fine tune if needed.

• Pixel-reassigned ISM imaging with up to 1.7× higher resolution
• Computational sectioning removes out-of-focus light for cleaner data
• Deconvolution on in-focus components for sharper structural detail
• Clear visualization of ISM, sectioning, and deconvolution improvements

• One-click deconvolution with minimal user interaction
• Fast, intuitive processing for ISM and ISM-FLIM datasets
• Batch analysis for series measurements and multi-ROI workflows
• Seamless operation within the Luminosa environment for immediate feedback

• Simultaneous computational sectioning and lifetime-species separation
• Access to raw time tags from all 23 PDA-23 SPAD pixels
• Fine-tuning options for sectioning and deconvolution parameters
• Extensive export formats for images and graphs (BMP, PNG, OME-TIFF, SVG)