Software

NovaISM NEW

Image Scanning Microscopy Analysis Software

  • Higher resolution than confocal imaging with pixel-reassignment and one-click deconvolution
  • State-of-the-art computational sectioning in combination with lifetime species separation, improving contrast and resolution
  • Seamless integration with Luminosa – Optimized for the PDA-23 SPAD array detector
  • Comprehensive data export options – Supports multiple formats, including OME-TIFF and scalable vector graphics
Image NovaISM Image Scanning Microscopy Analysis Software
Confocal fluorescence image at 640 nm excitation. Limited resolution results in blurred and overlapping signal spots. Enhanced fluorescence image after computational sectioning and deconvolution

NovaISM is a powerful Fluorescence Lifetime Imaging Microscopy (FLIM) analysis software specifically optimized for the PDA-23 SPAD array detector. It combines cutting-edge pixel reassignment, computational sectioning, and deconvolution techniques to achieve up to 1.7 times higher resolution compared to conventional confocal microscopy images.

By effectively rejecting out-of-focus light, NovaISM enhances the signal-to-noise ratio and significantly improves the lifetime contrast of FLIM images. These advantages enable faster image acquisition or gentler imaging of live samples, making it an essential tool for advanced image scanning microscopy.

screenshot of intensity profile

Advanced ISM-FLIM Analysis: High-Resolution Imaging with Deconvolution

Quality and precision you can trust

  • Computational sectioning algorithm in combination with lifetime-species separation
  • Transparency over the parameters used for deconvolution and computational sectioning

Save time and simply focus on your samples

  • One-Click Deconvolution
  • Minimal user interaction required
  • Efficient analysis of series measurements, including batch analysis and parameter plots of multiple-ROIs

 

Advanced flexibility

  • Raw data include individual time tags of each of the 23 pixels of the PDA-23 SPAD array
  • Many data export options for graphs and images (BMP, PNG, OME-TIFFf, Scalable Vector Graphics)

Species separation after ISM

Images with more resolution and higher contrast

Follow the effects of ISM, computational sectioning, and deconvolution step-by-step

Sum of all 23 pixels (pseudoconfocal)
Intensity image from summing up all 23 pixels of PDA-23 Detector. This corresponds quite well to the usual confocal imaging.
U2OS Cells by GattaQuant, Tubulin staining with Alexa488.

Pixel reassignment: ~ 30% more resolution
Intensity image obtained for Image Scanning Microscopy algorithm (pixel re-assignment) offering ~ 30% more resolution in comparison to the confocal case.

Computational sectioning, reject out-of-focus light
Intensity image after ISM and Computational Sectioning. The Computational Sectioning rejects out-of-focus light resulting in an image with higher contrast.

Result: Deconvolution: ~ 30% more resolution
Intensity image after ISM and Computational Sectioning and Lucy-Richardson Deconvolution. The deconvolution increases the resolution about ~ 30%.

 

Advanced Computational Sectioning: Remove Out-of-Focus Light and Separate Lifetime Species

The computational sectioning is based on an unmixing of the intensity distribution over the PDA-23  detector array.  This process allows for rejection of light coming from out-of-focus planes which in thicker samples can be problematic as it limits the contrast, produces artifacts in the deconvolution process and interferes in the lifetime –species determination.  With computational sectioning offered by NovaISM all these problems are resolved. 

Standard fluorescence image
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.

Computational Sectioning: Separating in-focus and out-of-focus signal components for improved image clarity and lifetime species separation.
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.

Refined ISM-FLIM Image: Enhanced resolution and contrast after computational sectioning and lifetime species separation.
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.

principle of computational sectioning
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.

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Licence type License available for either 1, 2, or 4 years
Data formats Available for *.spqr files and *.ptu files acquired with the PDA-23 SPAD Array Detector in Luminosa
PC system requirements
  Minimal Recommended
Operating system Win10 64-bit Win10 / Win11 64-bit
CPU Intel Core i3-8100 or AMD Ryzen 3 1300X  Intel Core i7-13700 or AMD Ryzen 7 5700X
RAM  8 GB  32 GB
GPU AMD Radeon RX550/550 Series or Nvidia GeForce GT 1030 AMD Radeon RX 6700 XT or Nvidia GeForce RTX 3060
Screen resolution QHD (2560 × 1440) 4K (3840 × 2160)

All Information given here is reliable to our best knowledge. However, no responsibility is assumed for possible inaccuracies or omissions. Specifications and external appearances are subject to change without notice.

  Luminosa NovaFLIM NovaISM NovaFLIM+NovaISM
InstaFLIM
  • Fast GPU accelerated analysis with minimal user interaction
  • 4 different FLIM contrast options: Fast lifetime, multi-exponential decay fit, phasor plot, and pattern matching
  • Suggestion for FLIM species separation
  • Suggested parameters can be fine-tuned
  
Phasor plot for marker multiplexing    
Multi-frame analysis    
Batch analysis of series measurements (time-lapse,z-stacks, tiling, …)    
Advanced ROI handling, multi-ROIs, import of external ROIs    
Parameter plots of fitted parameters over multiple ROIs    
Representation of the image information as 
user defined 1D and 2D histograms of fitted parameters		    
ROI selection via parameter histograms    
User configured analyses  allowing for
FLIM, FLIM-FRET, steady-state FRET and anisotropy imaging		    
Extended export options  of graphs and images     
Image Scanning Microscopy-FLIM 
(Resolution enhancement via a pixel reassigned image 
available in combination with PDA-23 Add-on)		  
Re-assignement vectors calculation from image  
Resolution enhancement Deconvolution for ISM-FLIM (enhanced FLIM contrast via the rejection of contribution from out-of.plane light; Available in combi with PDA-23 Add-on)    
Computational Sectioning in combination with lifetime-species seperation (enhanced FLIM contrast via the rejection of contribution from out-of.plane light; Available in combi with PDA-23 Add-on)    


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