AxioCam + HXP 120 vs PCO edge + Lumencor
|Warm up period||10 min||Less than 1 minute|
|Shutter||6ms||Less than 0,1 ms|
|Stability of the light intensity||It could vary by time||Stabile|
|Lamp type||Mercury||LED technology|
|Brightness control||manual control||software-controlled|
|AxioCam MRm - HXP120||PCO edge - Lumencor|
|Scanning time in 20x||7 min 43 sec||2 min 56 sec|
|Scanning time in 40x||21 min 52 sec||5 min 46 sec|
The advantege of PCO edge + Lumencor
• multiband filter support
• bleanching reduction (the tissue on the slide is illuminated only for the time of exposition)
• higher brightness, lower exposition time
• 16 bit picture versus 12 bit of AxioCam
Our technology allows that our scanner could scan only the area of the tissue, which was detected on the preview image, this speed up the scanning process verus the competitors where the whole slide is scanned, regardless the tissue size on the slide. In brightfield view it is easier to detect the tissue as it is clearly visible on the slide, however in Fluorescence mode this conventional method would not work as it not possible to detect the tissue on the slide. 3DHISTECH developed the dark-field preview which detects the tissue on the slide thanks to the oblique illumination.
An example of normal and dark field preview of a FL sample:
The method works for both brightfield and fluorescent samples. This option is recommended for you to choose in case the sample is not correctly stained (for example, the specimen is lacking contrast) when using Brightfield illumination.
FISHQuant 1.15.4. sp2.
The new FISHQuant software with new algorythms, options and new design will certainly impress its users.
On which samples can the FISHQuant be used?
• Tissue sections
• Cytology samples: smears, cell suspensions, cell cultures
The new cell segmenting algorythms allow for the detection of nuclei in tissue and cytology samples. Metaphases in cell cultures are evaulated alongside interphase cells.
Microscope Image Segmentation Profiles (MISP) are used which store the segmenting paramters and the probe type. These profiles can be used on identically stained samples for automatic evaluation.
New probe definition process
Probes can be defined using the existing probe rules of manufacturers. Clustering the nuclei will be based on this probe definition in tissue and cytology samples.
New Profile Setup
FISHQuant uses an automatic nuclei and spot detection algorythm for all sample types. A manual set up including nuclei size definition, separate spot treshold definition for each used channel is also available. Quick measurement functionality allows for nuclei far apart to be hand picked and included into the measurement one by one.
Spot number calculation
If the sample is highly/major amplified, then the area of the indistiguishable signal is devided by the gene signal average of the non amplified nuclei to give a good approximation of the spot numbers and calculate the magnitude of the amplification.
Optional settings to help measurement
Option to stop the analysis if the other/artifact cluster reaches a user set percentage. An error message will apear to indicate a possible wrong probe type setup or that the sample does not contain the specific genetic alterations. Option to automatically stop analysis if the positive nuclei count reaches a certain number.
An automatic second measurement can be saved in the MISP. Many tumors can contain nuclei with non specific deviation and the ploydity of the cell can change. This would be indicated by a numerical deviation. The second measurement clusters the anaploid and polyploid cells.
New Data Visualization Tool
• Cell relocation on the Virtual slide: All segmented nuclei can be visualized in a gallery and on the slide simultaneously
• Visualization by color channel
• Interactive filter table to organize visualization options based on cluster results. The clusters turned on will apear in the gallery and the user selectable piechart or bar diagram.
• Turning on and off the clusters on the diagrams also changes the content of the gallery accordingly.
• Manual re clustering in all measurement modes.
• All changes can be saved reflecting statistical changes as well.
• Export your data into Excel
• Generate a Measurement report.
Unique Secondary measurement and its visualization
It is possible to run a second, numerical deviation measurement for each spot channel to separate the tumor cells into aneoploid or polyploid clusters.
The two measurements can be visualized together in one new diagram called HistoPlot. A 2Dimensional scatter plot and histogram.
• The y axis shows numerical deviation clusters, x axis shows the first measurement clusters.
• The dots indicating the cells in the scatter plot can be visualized.
• Whole clusters can be turned on and off
• All data for the cluster/nuclei is shown in a corresponding tooltip
• It is possible to rescore cells. These changes are then saved in the slide.
• Both primary and secondary measurements can be exported to Excel.
Cy3, Cy5, FITC, TexasRed, Rhodamine, FITC, Aqua, Gold, DEAC (418-467 nm), DAPI