Vectra Polaris slide scanner; digital pathology


Location: Robert Kilpatrick Clinical Science Building, room 533
Contact: Dr Kees Straatman (krs5)


Phenochart free slide viewer for the Vectra Polaris can be installed on your own computer via the University's Software Centre or the latest version can be downloaded from the Akoya Bioscience website.

The AIF has purchased a PerkinElmer (now Akoya Biosciences) Vectra Polaris slide scanner with funding from the University Research Equipment and Infrastructure Fund, LD3 and AIF. The system has been installed in RKCSB room 533 but all users will have access to the system. An extra software licence for the Inform analysis software will be installed on one of the analysis computers in the Henry Wellcome Building.

The Vectra Polaris can be used for bright field slide scanning, multi-colour fluorescence slide scanning and multi-spectral slide scanning. See for adiitional information http://www.perkinelmer.co.uk/lab-solutions/resources/docs/PRD_Vectra-Polaris_013272_01.pdf.

Using multi-spectral imaging the system can separate up to 9 different colours as well as separating autofluorescence background from the real signal. The system can also be used with filter cubes to separate up to 5 different colours.

The system is optimized for Akoya Biosciences Opal assay kits (https://www.akoyabio.com/phenopticstm/assay-kits-and-reagents) but other fluorescent markers can be used as well.

For automatic identification of specific tissue types we have obtained 3 licenses for the integrated inForm image analysis software. One license is installed on the system, one license is with the LCRC and one will be installed in the Henry Wellcome Building image analysis room.

Vectra Polaris example

Figure 1; Breast cancer tissue labelled with the Opal 7-Color Multiplexed IHC Kit. Left to right: spectrally unmixed composite, tissue-segmented image, cell-segmented image and overlaid phenotyping of each cell obtained using the Inform analysis software (source: PerkinElmer).

Pricing

Planned pricing system (this is independent from the normal AIF charges, so will be charged extra!!); will potentially be adjusted after we have some more experience with the system:

For small sample size £15.00 per hour (internal use; subject to change). Large projects please contact krs5.

External users welcome. Please contact krs5@le.ac.uk for availability and pricing.

Specifications

Available magnification:
10x/NA = 0.2
20x/NA = 0.45
40x/NA = 0.75

For available filter combinations and spectral information download the filter cube details.

Filter cube reference list:
1. Opal 480 similar to coumarin
2. Opal 520 similar to Alexa488, FITC, Cy2, GFP
3. Opal 570 similar to Alexa555, Cy3
4. Opal 620 similar to Alexa594, Texas Red-X, RFP
5. Opal 690 similar to Alexa660-700, Cy5.5
6. Opal 780 similar to Alexa750, Cy7

Synthetic database

Below an overview of the dyes included in the synthetic database

fluorescent dyes included in the Vectra Polaris synthetic database  Synthetic database-2

Sample preparation

Use coverslip number 1.5

Don't use mounting media containing DAPI but use a DAPI staining step before you mount your samples.

Don't use nail varnish to seal your coverslip. The Vectra Polaris has to auto-detect your sample and will use the coverslip edges to detect the area for scanning. Nail varnish might cause the system to fail to detect the coverslip edge. As a result you need to use a hard-setting (curing) mountant and PerkenElmer/Akoya Biosciences advices to use ProLong Diamond from Invitorgen/ThermoFisher (no commercial interest from AIF) for the Opal dyes, but other curing mountant will probably work as well as long as they can be combined with your fluorescent markers.

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