TissueGnostics, the expert in cell counting and intensity measurement, has released the new HistoQuest Software. In it’s 3rd generation HistoQuest continues the advance of FACS-like data analysis and integrates it into an outstanding and intuitive graphical user interface.
HistoQuest uses patented cell-identification algorithms for nuclear segmentation. The intuitive user interface and automated Cut Off Suggestion in combination with the unique Back- and Forward- Connection is an ideal tool for data analysis and data control.
Immunohistochemistry images must be colour separated for analysis. HistoQuest provides two modes for this – fully automated colour detection for up to three differing colours (counterstaining and two markers) or semi-manual separation for several differing colours.
While the fully automated mode is very fast, the semi-manual mode allows for more fine-tuning. The more differing the colours will be, the better the results.
For a differentiated analysis, Regions of Interest and Exclusion Regions can be drawn on the slide. These shapes can be coloured by the user. Annotations can be made in various forms.
HistoQuest 3.5 permits the analysis of projects scanned with TissueFAXS/HistoFAXS systems, but also of images of samples scanned with other microsocopes. Digital slides from Mirax, Pannoramica and Hamamatsu scanners can be imported for analysis.
The analysis of TMA is fully supported. Projects scanned with TissueFAXS/HistoFAXS systems already have TMA spots named in columns within the blocks. For users needing to analyse imported TMA projects from other scanners automatic or manual spot ID capabilities can be integrated into the software.
HistoQuest 3.5 permits loading more samples of the same type into the analysis project (up to 12, depending on sample size) and analyzing them with the settings elaborated in the first experiment. Profiles for colour separation and segmentation settings can be reused to analyse further samples as well.
Several new parameters such as Calculated Parameters to measure ratios and the Percentile measurement parameter for intensely stained areas within Measurement Masks are available.
Comparative analyses of consecutive sections of tissue stained with different markers on the same tissue areas can be done by using Image Compare Sets.
Comparison of graphical data representations (scattergrams, histograms) can be done side by side or in overlays using the Compare Set feature.
Three data export modes are available to export analysis data. The Raw Data List feature is a list based feature which displays data for all analysed objects.
The Print Report feature provides a fast overview of a project and shows all relevant graphics and statistics in a report. This report can be exported in a variety fo formats. The Statistic Report tool allows to pick out specific sets of data, even across analysis projects.
HistoQuest streamlines all features needed from optimal viewing to accurate image processing as well as batch processing and reporting. HistoQuest is built to help you enjoy your work. It offers optimal overview of up to 6 different slides for visual inspection and at the same time a set of precise image analysis algorithms to prepare a quantitative report in a very user friendly but highly accurate and secure form.
TissueGnostics has made major efforts to equip its users with Software that is tailored to their needs. As the final result HistoQuest 3.0 is the first HistoQuest to be modularized. Taking into consideration the different applications of image analysis software around the globe, TissueGnostics defined 2 major classes, with altogether 6 module-packages:
This entry module provides the capability to import original brightfield color-images and draw individualised regions of interest (ROI's) to measure stained areas in a sample. It also features an annotation tool. Marker separation is semi-manual using colour picking in a multiple shading mode. Data display is in histograms and raw data can be exported to Excel or PDF.
Stand-alone or incombination with any other module
Similar to the histology module, the nuclear counting module additionally allows users to quantify stained nuclei and measure several analytical parameters, such as area and staining intensity of nuclei.
Basic module for the modules 3 to 5
This module adds more cell-structured analysis to the entry modules: ring mask around the nucleus and cell mask detection of cytoplasmatic markers is available through a growth algorithm. Data can now also be displayed in scattergrams.
Add-on to anterior modules
This module additionally provides automatic color separation (single shade mode), virtual channels for comparative measurements in nuclear versus cytoplasmatic compartments, and an illumination correction tool. Compare sets allow the graphical comparison of diagrams. Reloading of marker and experiment profiles, and a report generator are also provided. Tools such as a ruler and a scale bar make exact sample measurements possible. Propagation & grouping of ROI's makes analysis of consecutive cuts simpler and faster.
Add-on to anterior modules
The entire features and capabilities (excluding TMA analysis) of the Software are provided: in addition to the diagnostics module, further automation is possible using tissue detection and a cutoff suggestion for the separation of positive and negative populations in the diagrams. Scattergram and histogram results can be controlled visually by using the forward and backward connection in conjunction with images. The images can now also be imported from specific scanners and be compared using the multi-sample-viewer. A manual correction tool allows manipulation of the identified nuclei and specified regions can be excluded from analysis. Finally, ROI's and annotations can be produced quicker and more extensively with the easy-ROI tool.
Add-on to anterior modules
This module provides full analysis capabilities including Tissue Microarrays. TMA scanning projects from the TissueGnostics product line TissueFAXS/HistoFAXS can be imported with each spot, block or slide already identified and named. The spots in TMA images from other systems can be identified with several features, allowing for automatic detection or manual selection.
Add-on to any other module
The brand new Multi Tissue Viewer aligns up to six consecutive sections of one sample, with six different stainings, for optimal comparison of analysis results in the same anatomical structure.
HistoQuest can load complete HistoFAXS projects and display the overview of the regions of interest.
Graphical tools can be used to draw analysis selections on these areas of interest for subsequent HistoQuest analysis (e.g. for tumor areas, etc.).
Analysis selections can be linked to form a compound analysis selection and can also be inverted.
Alternatively, series of images from other sources can be imported.
As the spatial relation is not available, such image series cannot be reassembled into an overview.
They can, however, be analyzed by HistoQuest.
Contrary to immunofluorescence, the marker information in immunohistochemistry images is contained in one image – in order to analyse it, a color separation has to be done. Information for the optimization of the picked color values can be gained by very rapid interim result calculation.
In HistoQuest, this is done semiautomatically. RGB color values are manually picked with a color picker tool.
The automatic segmentation which follows produces a gray value channel image for each marker, allowing treatment with the TissueGnostics algorithms.
HistoQuest provides several possibilities for color separation. Depending on experimental aims different separation methods can be used:
Automatic nuclear segmentation is achieved via TissueGnostics patented set of algorithms.
Once rapid test calculations done on representative images have established a good segmentation, nuclear segmentation as well as the measurement masks can be automatically computed for virtually any number of corresponding images.
Nuclear segmentation in HistoQuest is completely automatic after the input of a few starting values:
Evaluate your result by selecting any event or group of events in the scattergram.
The corresponding cell will be immediately highlighted in the image, allowing the user to control reliability of the analysis data.
The real-time back gating feature from scattergram quadrants and gates is used for interactive Cut Off setting.
Attention: Consult operating instructions for use!