Vectra Polaris (UNSW & Ramaciotti)

Vectra Polaris

The Vectra Polaris Automated Quantitative Pathology Imaging System is a tissue imager for digitalising and analysing paraffin embedded or frozen tissue sections, but also Tissue Microarrays (TMAs). It can scan in Brightfield or widefield fluorescence, but it also integrates the power of multispectral imaging to allow up to 7 colours on a whole slide scan or generate spectrally unmixed regions for up to 9 colours on the same tissue, using liquid crystal tuneable filter technology.  

The files can be viewed and annotated with the free software Phenochart and quantitatively analysed with the licenced Inform® software, or open source QuPath software, both designed for phenotyping individual cell types, counting cells, analising morphology… 

 

More information can be found here: 

https://www.perkinelmer.com/lab-solutions/resources/docs/PRD_Vectra-Polaris_013272_01.pdf 

Microscope body:
Upright
Motorised Stage:
Yes
Temp Control:
No
CO2:
No
Software:
Vendor software

Common Applications

Digitalization of microscope thin tissue section (paraffin/frozen) 

Quantifying biomarkers in tissue section: 

  • Phenotyping immune cells for cancer immunology research 

  • Transduction signaling pathway activity (pAKT, pERK, pS6, p13/mTOR, MAPK or EGFR 

  • Apoptosis and/or proliferation assay 

  • Necrosis and fibrosis using conventional stains 

  • Cell cycle characterization 

  • Inflammation 

  • Lymph node metastasis 

  • Autoimmune diseases 

  • Transplant acceptance 

  • Neurodegenerative diseases 

  • RNA-ISH 

  • Tumor microenvironment exploration 

Brightfield imaging (RGB images)  Brightfield slide scanning imaging can only be used for histology-stained thin sections (about 4 um) as the technique does not give a lot of contrast on its own. RGB images are generated to reflect the proportion of different chromophores in the samples.  
Widefield Fluorescent Imaging (Monochromatic channels)  Widefield Fluorescent Slide Scanning allows to use fluorescent markers to highlight targets of interest. Each channel reflects the intensity of the signal fluorophores captured by the monochromatic camera. This slide scanner can separate up to 7 colours and 9 colours if using spectral unmixing and liquid crystal tuneable filters. Please note that a careful selection and optimisation of the fluorophores used must be done prior to multiplex slide scanning (please contact us before buying any reagents). 

Objective Lenses

Magnification 

Numerical Aperture 

Corrections 

Immersion 

MISC. 

10x 

0.2 

PL APO objective 

Dry 

  1. um/pixel 

20X 

0.45 

PL APO objective 

Dry 

0.5 um/pixel 

40X 

0.75 

PL APO objective 

Dry 

0.25 um/pixel 

Light Sources

Source 

Wavelengths 

Misc. 

UV 

365 nm 

 

Violet 

405 nm 

 

Violet 

440 nm 

 

Blue 

475 nm 

 

Cyan 

510 nm 

 

Green/Yellow 

550 nm 

 

Red 

630 nm 

 

Far Red 

660 nm 

 

Filters

 

 

 

 

 

7 Color Whole slide unmixing 

 

 

Based on Opal 7 color kit 

Name 

Excitation 

Emission 

Example of compatible Fluorophores 

Multi band A 

DAPI/Opal 570/Opal 690 

375-407 

526-551 

624-674 

438-468 

561-581 

689-729 

DAPI/Hoechst 

Alexa 555/Cy3 

Alexa 660/700/Cy5.5 

Multiband B 

Opal 480/Opal 620/Opal 780 

403-445 

578-603 

715-765 

458-498 

613-653 

765-900 

Coumarin 

Alexa 594/Texas Red-X/RFP 

Alexa 570/Cy7 

Multiband C 

Opal 520 

471-501 

523-541 

Alexa 488/ FITC/Cy2/GFP 

 

 

9 Color field of view unmixing 

 

 

Based on Opal 9 color kit 

DAPI + Opal 780 

381-393 

715-755 

409-700 

765-850 

 

FITC 

460-500 

LP 510 

 

Cy3 

532.5-557.5 

LP 570 

 

Texas Red 

567-589 

604-710 

 

Opal 480+Cy5 

403-445 

590-650 

455-580 

662-738 

 

*LP: Long pass 

 

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Detectors

Detector 

Sensor type 

Pixel Format 

Pixel size 

Description 

8/10-bit monochrome TDI line-image capture camera 

Single line CCD 

Depend on the scan area 

20x: 0.50 µm/pixel  

Scanning region: 23.4 x 55 mm 

Miscellaneous

  • Image File Formats: QPTIFF 

  • Optimized coverslip thickness: #1 or #1.5 

  • Use only standard pathology glass slides (75 x 25 x 1 mm) 

  • It’s highly recommended to use DAPI or Hoechst in your staining to allow the automatic recognition of the focal plan of your sample. 

  • Do not use mounting media containing DAPI, but stain DAPI or Hoechst as a separated step before mounting the coverslip with preferably prolong gold diamond (or a mounting media that has a refractive index of 1.51 and preserve fluorescence).  

  • Use only hard-set mounting medium.  

  • Do not use nail polish to seal the samples as it affects the automatic recognition of the edges of the coverslip.