Aperio at2 digital scanner

Manufactured by Leica

Sourced in United Kingdom, Israel, United States

The Aperio AT2 digital scanner is a high-performance scanner designed for the digitization of pathology slides. It captures high-resolution digital images of microscope slides, enabling efficient storage, retrieval, and analysis of pathological samples.

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Lab products found in correlation

Bond polymer refine detection system, by Leica (2 mentions) Dibutylphthalate polystyrene xylene (dpx), by Merck Group (2 mentions) Imagescope, by Leica (2 mentions) Direct red 80, by Merck Group (2 mentions) Bond 3, by Leica (2 mentions) Hpa 019 051, by Merck Group (1 mentions) Sensifast probe lo rox kit, by Meridian Bioscience (1 mentions) Anti ki67 antibody, by Leica (1 mentions) Anti mdm2 antibody, by Abcam (1 mentions) Anti p53, by Abcam (1 mentions) Hematoxylin, by Bio-Optica (1 mentions) Aperio image analysis software, by Leica (1 mentions) Eosin g, by Bio-Optica (1 mentions) Phosphate buffered saline (pbs), by Merck Group (1 mentions) Hematoxylin and eosin, by Merck Group (1 mentions) Hematoxylin, by Merck Group (1 mentions) Axio imager m2m microscope, by Zeiss (1 mentions) Aperio imagescope, by Leica (1 mentions) Mayer s hematoxylin, by Bio-Optica (1 mentions) Eukitt, by Bio-Optica (1 mentions)

Close spelling variants of this product

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13 protocols using aperio at2 digital scanner

Histological Analysis of Colonic Inflammation and HIF-1α Expression

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Colons were collected from sacrificed mice at the end of each treatment schedule, as described above, fixed in 10% neutral-buffered formalin, and embedded in paraffin. Serial 5 µm microtome-cut sections were then stained with hematoxylin and eosin (Sigma-Aldrich). Slides were acquired with an Aperio AT2 digital scanner (Leica Biosystems) and analyzed with an Aperio Imagescope (Leica Biosystem). Scoring of colonic inflammation was determined as reported by Nagahama et al.^{44 (link)} with minor modifications, by assessing mucosa thickening, inflammatory cells, and submucosa cell infiltration. Each criterion was scored as 0–4, and the sum of each score was defined as the histological score.
Immunohistochemical staining of HIF-1α was performed on colon sections of mice with the anti-HIF-1 alpha antibody (Abcam ab2185, 1:2000) and antigen unmasking in citrate buffer. Sections were incubated with specific Detection KIT-HRP (Abcam, ab236469) according to the manufacturer’s instructions, then counter-stained with hematoxylin to visualize nuclei, dehydrated and mounted in DPX (Sigma-Aldrich). Bright-field images were acquired with an Aperio AT2 digital scanner (Leica Biosystems) and analyzed with an Imagescope (Leica Biosystem) or using a Zeiss Axio Imager M2m microscope with a ×10 or ×20 objective.

Torretta S., Scagliola A., Ricci L., Mainini F., Di Marco S., Cuccovillo I., Kajaste-Rudnitski A., Sumpton D., Ryan K.M, & Cardaci S. (2020). D-mannose suppresses macrophage IL-1β production. Nature Communications, 11, 6343.

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Quantifying CCDC6 Protein Expression in Ovarian Cancer

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A case series of 251 primary ovarian cancers was analysed on TMA. 6 TMAs were prepared with all the analysed samples [33 (link)]. Each tissue was examined for representability, and subsequently immunostained with anti-CCDC6 antibody (HPA 019,051, from Sigma-Aldrich), CCDC6 immunoreactivity of tumour cells was annotated. Visual annotation included staining intensity (negative, weak, moderate or strong), fraction of stained cells (% of total counted tumour cells), and subcellular localization. Following acquisition of anti-CCDC6 immunostained glass slides with a digital scanner, we also calculated the H-score on digital images by QuPath image analysis software [34 (link), 35 (link)]. Immunohistochemistry was performed as already described [30 (link)]. Median H-score was 64.2 (IQR 28.8; 97.2); Mean H-score was 67.5 (sd 47.1). The frequency distribution of digitally evaluated CCDC6 H-Score is summarised in Additional File 3: Table S2. A summary of study population is shown in Additional File 4: Table S3.
PDX FFPE sample slides were immunostained with anti-CCDC6 antibody (HPA 019051, from Sigma-Aldrich) and IHC signal was quantified on digital slides with Positive Cell Count function of QuPath software. CCDC6 protein expression was reported as optical density arbitrary units. All the glass slides were digited with Aperio AT2 digital scanner (Leica).

Morra F., Merolla F., Damia G., Ricci F., Varricchio S., Ilardi G., Arenare L., Califano D., Napolitano V., Fruscio R., Melillo R.M., Palazzo L, & Celetti A. (2022). The disruption of the CCDC6 – PP4 axis induces a BRCAness like phenotype and sensitivity to PARP inhibitors in high-grade serous ovarian carcinoma. Journal of Experimental & Clinical Cancer Research : CR, 41, 245.

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Quantifying vgr and bol Gene Expression in WCR

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The expression of vgr and bol gene was quantified from WCR eggs, neonates, 1^st, 2^nd, 3^rd instar, pupae, and adults after feeding on diet incorporated with 50 ng μl⁻¹ or different doses of vgr and bol fragment dsRNA. The designs of primers and probe regions are listed in Supplementary Table 6. Gene expression was analyzed using one-step real-time qRT-PCR. The assay was run, with 3 replicates per sample, using a single-plex set up with Bioline Sensifast Probe Lo Rox kit (Taunton, MA) and analyzed using the 2^−ΔΔCt method based on the relative expression of the target gene and reference gene dvrps10. For in situ hybridization (ISH) analyses, target probes were designed by Advanced Cell Diagnostics (Hayward, CA) (listed in Supplementary Table 6). Insect samples were fixed in 10% neutral buffered formalin (4% formaldehyde) for 48 to 72 h and processed as previously reported^{11 (link)}. Slide images were acquired using a Leica Aperio® AT2 digital scanner and captured at 40x magnification with a resolution of 0.25 µm pixel⁻¹.

Niu X., Kassa A., Hu X., Robeson J., McMahon M., Richtman N.M., Steimel J.P., Kernodle B.M., Crane V.C., Sandahl G., Ritland J.L., Presnail J.K., Lu A.L, & Wu G. (2017). Control of Western Corn Rootworm (Diabrotica virgifera virgifera) Reproduction through Plant-Mediated RNA Interference. Scientific Reports, 7, 12591.

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Immunohistochemical Analysis of Ki-67, MDM2, and p53

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Three 2.0 mm diameter core biopsies were selected from the paraffin-embedded tissue blocks. The cores were transferred to tissue microarray (TMA) and the pathologists were blinded to the identity of the TMA slides as previously described [19 (link)]. IHC tests with a rabbit monoclonal anti-Ki-67 antibody (1:1000, LEICA), an anti-MDM2 antibody (1:300, Abcam), and an anti-p53 (1:400, Abcam) antibody were performed using the Leica Bond-III system (Aperio, CA, USA). The expression of slides was examined by an Aperio AT2 digital scanner (Leica Biosystems). The H-score was obtained by multiplying the staining intensity by a constant to adjust the mean to the strongest intensity [H-score = 3 × (percentage of strong staining)] (1.0%, weak; 2.0%, moderate; 3.0%, strong) to give a score ranging from 0 to 300.

Yao X., Liu Q., Zhao S., Cheng R., Liu C, & Zhang G. (2023). Expression and Clinical Significance of MDM2 in Non-Functioning PitNETs. Medicina, 59(2), 373.

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Immunohistochemistry Protocol for TMAs

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Before conducting immunohistochemistry (IHC), the TMA slides underwent staining with hematoxylin and eosin and were assessed for both quality and tumor content. The TMAs were then processed using the Leica BOND‐III, an automated, random, continuous‐access slide‐staining system located in Nussloch, Germany. This system allowed for the simultaneous execution of multiple IHC assays. To detect the primary antibody, the Bond Polymer Refine Detection system from Leica Biosystems was employed. The resulting immunostained slides were subsequently examined for expression using the Aperio AT2 digital scanner, also from Leica Biosystems. Staining intensity was categorized into four distinct levels: 0, negative; 1+, weak; 2+, moderate; and 3+, strong. The percentage of immunostaining was recorded and H‐scores were calculated as:

H - score = (% cells 1 +) + 2 * (% cells 2 +) + 3 * (% cells 3 +)

The maximum H‐score was 300, corresponding to 100% of cells stained with strong intensity.

Fang Q., Liu C., Nie D., Guo J., Xie W, & Zhang Y. (2024). Phosphorylation of PBK at Thr9 by CDK5 correlates with invasion of prolactinomas. CNS Neuroscience & Therapeutics, 30(2), e14629.

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Single-plex Chromogenic IHC for PD-L1

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Single-plex chromogenic IHC was performed using an automated staining system (Ventana BenchMark) with PD-L1 SP263 clone dispensed neat via a locked-in protocol, as recommended by the company (Ventana, CC1 pre-treatment for 64 min, Ventana Optiview detection protocol), a diaminobenzidine (DAB) reaction was used to detect antibody labelling with haematoxylin counterstaining. To assess specificity and sensitivity, an intra-run reproducibility section from a four core TMA was used in each test run, representing PD-L1 expression levels of <1%, 1–49%, and >50%, as well as a positive control (tonsil). All clinically assessed cases were scored by teams of two individuals who received training and are certified competent for PD-L1 scoring. All DAB IHC slides were scanned at 40x on an Aperio AT2 digital scanner (Leica Biosystems, Milton Keynes, UK). Images were automatically stored on a secure network server.

Abdullahi Sidi F., Bingham V., Craig S.G., McQuaid S., James J., Humphries M.P, & Salto-Tellez M. (2020). PD-L1 Multiplex and Quantitative Image Analysis for Molecular Diagnostics. Cancers, 13(1), 29.

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Histological Analysis of YAP and Ki67 in Kidney Sections

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For histological analysis mice were sacrificed at the indicated time, kidneys were collected, weighted and fixed in formalin 10% (BioOptica, #05-01005Q), included in paraffin and cut 5 μm/slides. Kidney sections were air-dried and rehydrated in PBS (Sigma-Aldrich, #P4417). For YAP staining kidney sections were incubated with YAP antibody and 5 min in Eosin G (BioOptica, #05-10002/L); For Ki67 staining kidney sections were incubated with Ki67 antibody and 2 min in Hematoxylin (BioOptica, #05-06015/L). Sections were then washed, processed through a dehydration alcohol scale and mounted in DPX (Sigma-Aldrich, #06522). For the semi-automated quantification, slides were acquired with Aperio AT2 digital scanner at magnification of 40 × (Leica Biosystems) and analyzed with Imagescope (Leica Biosystem). YAP and Ki67 nuclear staining per cyst was performed by Aperio Image analysis software (Leica Biosystems) and counted manually.

Nigro E.A., Distefano G., Chiaravalli M., Matafora V., Castelli M., Pesenti Gritti A., Bachi A, & Boletta A. (2019). Polycystin-1 Regulates Actomyosin Contraction and the Cellular Response to Extracellular Stiffness. Scientific Reports, 9, 16640.

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Quantitative Immunohistochemical Analysis

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Immunohistochemically stained slides were scanned using an Aperio AT2 digital scanner with a 40× objective (Leica Biosystems Inc., Buffalo Grove, IL). The images were analyzed using Visiopharm Digital Image Analysis (DIA) software (for Windows 7, version 6.9.1; Visiopharm, Hørsholm, Denmark). The cytoplasm was defined by outlining the nucleus with a system trained to digitally “paint” cell nuclei. The proportion of positively brown-stained cells was obtained using a predefined algorithm and optimized settings, as previously described [26 (link)]. The immunohistochemical score was expressed as the percentage of positive cells (possible range 0–100). The median values were used as cut-off values for discriminating between low and high expression of immunohistochemical staining. Cut-off values for CD68, CD163, VEGF-A, VEGF-C, and M2 ratio (CD163+/CD68+) were 8.70%, 10.12%, 7.42%, 3.09%, and 1.17 with high cytoplasmic staining, respectively.

Hwang I., Kim J.W., Ylaya K., Chung E.J., Kitano H., Perry C., Hanaoka J., Fukuoka J., Chung J.Y, & Hewitt S.M. (2020). Tumor-associated macrophage, angiogenesis and lymphangiogenesis markers predict prognosis of non-small cell lung cancer patients. Journal of Translational Medicine, 18, 443.

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Histological and Immunohistochemical Characterization of Tissue Sections

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Formalin-fixed paraffin-embedded consecutive sections (4 µm) were dewaxed and dehydrated by a graded decrease alcohol series and stained for histology or immunohistochemical characterization (IHC). For histological analysis in bright-field microscopy, slides were stained using standard protocols for H&E (using Mayer’s H&E; BioOptica) and Sirius red staining (Direct Red 80, Sigma) for collagen fibers. For IHC, slides were immunostained for anti-GFP mAb (Thermo Fisher Scientific) and anti-RFP mAb (Rockland) according to the manufacturers’ instructions. Control experiments showed that on day 8 after infection, no tdTomato-expressing HSV-1_TOM-OVA–infected cells were detectable in skin. After antigen unmasking with citrate buffer, pH 6.0, endogenous peroxidase activity was inhibited by incubating slides with 3% peroxidase water for 20 min at RT. Both mAbs were used at dilution of 1:200 and developed with EXPOSE rabbit-specific HRP/DAB detection IHC kit (ab80437). After immunostaining, DAB substrate chromogen was applied to sections for 5 min at RT and counterstained with Mayer’s Hematoxylin, dehydrated, and mounted with Eukitt (BioOptica). Proper external positive and negative controls were run simultaneously. Slides were acquired with Aperio AT2 digital scanner at magnification of 200× (Leica Biosystems). Cells in epidermis and papillary dermis were counted manually.

Moalli F., Ficht X., Germann P., Vladymyrov M., Stolp B., de Vries I., Lyck R., Balmer J., Fiocchi A., Kreutzfeldt M., Merkler D., Iannacone M., Ariga A., Stoffel M.H., Sharpe J., Bähler M., Sixt M., Diz-Muñoz A, & Stein J.V. (2018). The Rho regulator Myosin IXb enables nonlymphoid tissue seeding of protective CD8+ T cells. The Journal of Experimental Medicine, 215(7), 1869-1890.

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Immunohistochemical Analysis of Subcutaneous Tumors

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Formalin-fixed (HT501128, Sigma-Aldrich) paraffin-embedded sections of subcutaneous tumors were dewaxed and hydrated through a graded decrease alcohol series. For immunohistochemistry (IHC), slides were immunostained with the Automatic Leica BOND RX system (Leica Microsystems GmbH, Wetzlar, Germany). Antigen-retrieval was performed using sodium citrate buffer at 100 °C. Primary antibodies anti-Ki-67 (dilution 1/200; Ki-67 (D3B5) #12202, Cell Signaling Technology, Inc.) and anti-cleaved caspase-3 (1/400; Cleaved Caspase-3 (Asp175), #9661, Cell Signaling Technology, Inc.) were developed with Bond Polymer Refine Detection (Leica, DS9800). Bright-field images were acquired with an Aperio AT2 digital scanner (Leica Biosystems) and Aperio ImageScope software (v12.4.3.5008, Leica Biosystem).

Ricci L., Stanley F.U., Eberhart T., Mainini F., Sumpton D, & Cardaci S. (2023). Pyruvate transamination and NAD biosynthesis enable proliferation of succinate dehydrogenase-deficient cells by supporting aerobic glycolysis. Cell Death & Disease, 14(7), 403.

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