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IngMarie Olsson
Sofie Gustafsson (technician), Frank Hammar (technician), Emma Mattsson (technician) and Ann-Sofi Strand (technician).
(i) Production of tissue microarrays (TMAs) and cell microarrays (CMAs), (ii) handling of tissues (biobank material) and cells/cellines used for TMA production and protein extraction, (iii) sectioning of tissue- and TMA-blocks, (iv) quality control of TMA-block during sectioning, (v) scanning of immunohistochemically stained TMA slides (vi) export and processing of generated images from TMAs and (vii) administration and control of TMAx for automated image analysis.
Tissue specimens are selected from the pathology department and representative samples are histo-processed and stored as paraffin blocks. Areas of representative tissue containing appropriate cell populations are defined from a hematoxylin-eosin stained tissue section representing each donor tissue block through visual inspection under the microscope. For the CMA cell lines are cultured in vitro, harvested and dispersed in agarose.
An automated tissue arrayer, ATA-27, is used to subsequently collect cores of tissue from corresponding donor tissue blocks, and to assemble these into a recipient TMA block. Following sectioning, quality assured TMA sections are delivered to the Immunohistochemistry module.
The 8 TMAs used for the Protein Atlas contain 144 selected normal tissues representing 48 different tissue types and tumor tissues from 216 patients representing the 20 most common tumor types (17 x 12 replicates and 3 x 4 replicates). The CMAs contain cells from human cancer cell lines (47) and leukemia cells from blood samples (12). In total 708 spots of tissues and cells are immunostained and analyzed for each antibody. In addition, a specially designed test-TMA is produced for titration of immunohistochemistry protocols.
Immunostained TMA slides are scanned to generate high-resolution digital images. Automated scanning is performed using 20x (tissue) or 40x (cells) magnification. Images from scanned TMAs are separated into spot images and exported as individual TIFF files. Each TIFF file representing an immunostained tissue (1 mm diameter) or cell sample (0.6 mm diameter) is processed and stored in a database. The images representing immunostained tissue sections are used for manual analysis (pathology-based) of antibody staining and annotation using a web based annotation software. Analysis of immunoreactivity in cells and cell lines in the cell TMA is performed by automated image-analysis to generate non-subjective quantitative data from immunohistochemistry.
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