FFPE tissue blocks were collected from all patients (n=86). Sections of the FFPE blocks were stained with haematoxylin and eosin and representative tumor regions were marked by a pathologist (LH). To construct a TMA, three cores of every tumor tissue, 1.0-mm in diameter, were sampled into a donor block using the fully automated TMA Grand Master (Sysmex, Norderstedt, Germany). The cores were taken from both central and peripheral parts of the tumor to allow intratumor heterogeneity analysis. Subsequently, serial 4 um sections were cut for further analysis.
Tma grand master
Manufactured by Sysmex
Sourced in Poland
The TMA Grand Master is a high-performance automated system designed for molecular diagnostic testing in clinical laboratories. It automates the entire process of nucleic acid extraction, amplification, and detection, providing reliable and efficient results. The TMA Grand Master integrates seamlessly with Sysmex's comprehensive suite of laboratory solutions to support comprehensive diagnostic workflows.
Automatically generated - may contain errors
Lab products found in correlation
Envision target retrieval solution, by Agilent Technologies (2 mentions)
Pt link pre treatment module for tissue specimens, by Agilent Technologies (2 mentions)
Silanized slides, by Agilent Technologies (1 mentions)
Liquid permanent red, by Agilent Technologies (1 mentions)
Envision flex hrp magenta chromogen, by Agilent Technologies (1 mentions)
5 protocols using tma grand master
1
Tissue Microarray Construction from FFPE Blocks
2
Tissue Microarray Construction for Colorectal Cancer
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A tissue microarray (TMA) was constructed at the Department of Pathology, University Hospital Basel, from each tissue sample from 754 unselected, non-consecutive patients with primary CRC following approval by the Regional Ethical Committee (EKBB, Ethikkommission beider Basel, Switzerland). Formalin-fixed, paraffin-embedded tissue blocks were prepared according to standard procedures. Tissue cylinders with a diameter of 0.6 mm were punched from morphologically representative areas of each donor block and brought into one recipient paraffin block (30 × 25 mm) using a TMA-Grand Master® automated tissue arrayer (3DHisteck, Sysmex AG, Horgen, Switzerland). Each punch was made from the center of the tumor so that each TMA spot consisted of at least 50% tumor cells. The detailed construction technique was previously described by our group [44 (link),69 (link),70 (link)].
3
Immunohistochemical Analysis of ENO1 Expression
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Tissue microarrays (TMAs) composed of three 1.5 mm tissue cores from each tumor were automatically constructed (TMA Grand Master, Sysmex, Warsaw, Poland). Immunohistochemical analysis was performed using rabbit polyclonal anti-ENO1 antibody (dilution 1:500) on 4-μm-thick paraffin sections mounted on silanized slides (Agilent DAKO, Santa Clara, CA, USA). The slides underwent automated dewaxing, rehydration, and heat-induced epitope retrieval with EnVision Target Retrieval Solution (Agilent DAKO, Santa Clara, CA, USA) for 30 min at 97 °C in PT Link Pre-Treatment Module for Tissue Specimens (DAKO). Liquid Permanent Red (Agilent DAKO, Santa Clara, CA, USA) was utilized as a detection system. Human breast and pancreatic adenocarcinomas were stained as positive controls. Negative controls were processed using FLEX Rabbit Negative Control, Ready-to-Use (Agilent DAKO, Santa Clara, CA, USA) in place of the primary antibody.
Scoring of ENO1 immunostains was performed using the H-score [(percentage at 1+) × 1 + (percentage at 2+) × 2 + (percentage at 3+) × 3], which integrates the intensity and percentage of positive cells into a combined score. The median H-score (200) was used as a cut-off value for high (H-score > 200) and low ENO1 (H-score ≤ 200) expression [23 (link)].
Scoring of ENO1 immunostains was performed using the H-score [(percentage at 1+) × 1 + (percentage at 2+) × 2 + (percentage at 3+) × 3], which integrates the intensity and percentage of positive cells into a combined score. The median H-score (200) was used as a cut-off value for high (H-score > 200) and low ENO1 (H-score ≤ 200) expression [23 (link)].
4
Multifocal Ileum NET Profiling
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Following the initial re-evaluation, representative tumor areas from the tumor center and the invasive front from every suitable primary tumor that harbored a high burden of invasive tumor were marked on HE slides. Next, formalin-fixed paraffin-embedded (FFPE) tumor samples from each separate primary tumor of the multifocal ileum NETs were assembled into a tissue microarray (TMA) using a fully automated Tissue Microarrayer (TMA Grandmaster, Sysmex, Budapest, Hungary) with a core size of 1.5 mm.
5
Immunohistochemical Analysis of ABCA1 Expression
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Tissue microarrays (TMAs) composed of three 1.5 mm tissue cores from each tumor were automatically constructed (TMA Grand Master, Sysmex, Warsaw, Poland). Immunohistochemical analysis was performed using mouse monoclonal anti-ABCA1 antibody (clone HJ1 (ab66217), dilution 1:200, Abcam) on 4-µm-thick paraffin sections mounted on silanized slides (Agilent DAKO, Santa Clara, CA, USA). The slides underwent automated dewaxing, rehydration, and heat-induced epitope retrieval with EnVision Target Retrieval Solution (Agilent DAKO, Santa Clara, CA, USA) for 30 min at 97 ℃ in PT Link Pre-Treatment Module for Tissue Specimens (DAKO). EnVision Flex HRP Magenta Chromogen (Agilent DAKO, Santa Clara, CA, USA) was utilized as a detection system. Human normal liver was used as a positive control. Negative controls were processed using FLEX Rabbit Negative Control, Ready-to-Use (Agilent DAKO, Santa Clara, CA, USA) in place of the primary antibody. Scoring of ABCA1 immunostaining was performed using the H-score. The score is obtained by the formula: percentage of tumoral cells with weak immunoreactivity × 1 + percentage of tumoral cells with intermediate immunoreactivity × 2 + percentage of tumoral cells with high immunoreactivity × 3, giving a range of 0 to 300. The median H-score (200) was used as a cut-off value for high (H-score > 200) and low ABCA1 (H-score ≤ 200) expression [20 (link)].
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