Nanozoomer 2.0 ht slide scanner

Manufactured by Hamamatsu Photonics

Sourced in Japan, Germany, United States

The NanoZoomer 2.0-HT is a slide scanner designed for digitizing microscope slides. It features a high-speed scanning capability and can process multiple slides simultaneously. The device captures high-resolution images of the slide content, enabling digital archiving and analysis of the sample material.

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Lab products found in correlation

Ndp view2, by Hamamatsu Photonics (3 mentions) Entellan, by Merck Group (2 mentions) Ndp viewer software, by Hamamatsu Photonics (2 mentions) Bouin s fixative, by Electron Microscopy Sciences (2 mentions) Mz 16 stereomicroscope, by Hamamatsu Photonics (2 mentions) Ndp view, by Hamamatsu Photonics (2 mentions) Aperio imagescope, by Leica Biosystems (2 mentions) Ab32072, by Cell Signaling Technology (2 mentions) Leica dfc450 c, by Leica Microsystems (2 mentions) Histoquest version 6, by TissueGnostics (2 mentions) Pdgfra, by TissueGnostics (2 mentions) Hematoxylin, by Merck Group (2 mentions) Lsm 780, by Zeiss (2 mentions) Eclipse te300, by Nikon (1 mentions) Biotinylated secondary antibody, by Vector Laboratories (1 mentions) Vectastain dab kit, by Vector Laboratories (1 mentions) Icc50w microscope, by Leica camera (1 mentions) Vectastain abc kit, by Vector Laboratories (1 mentions) Cyclin d1, by Cell Signaling Technology (1 mentions) Lv1201, by Tissue Array (1 mentions)

82 protocols using nanozoomer 2.0 ht slide scanner

Histological Analysis of UCP1 Expression

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H&E staining or Oil red O staining was performed on formalin-fixed paraffin-embedded or OCT-embedded sections using a standard protocol, respectively. Brightfield images were scanned with the NanoZoomer 2.0-HT slide scanner (Hamamatsu, Japan) at different magnifications.
Immunohistochemical staining was performed on formalin-fixed paraffin-embedded sections using a standard protocol. Briefly, slides were dewaxed with xylene 3 times for 15 min each, hydrated through an alcohol gradient, and soaked in DPBS for 5 min. Antigen was recovered by pre-heating in the EDTA buffer (pH 8.0). Endogenous peroxidase activity was blocked with 3% H₂O₂ in PBS for 25 min. Non-specific binding sites were blocked using 3% bovine serum albumin in PBS. The slides were incubated overnight at 4 °C with rabbit polyclonal anti-UCP1 primary antibody diluted at 1:200 in PBS. According to the manufacturer’s instructions, the slides were rinsed in distilled water, followed by treatment with a secondary antibody (HRP labeled). Immuno-visualization was performed with 3,3’-diaminobenzidine (DAB) as substrate and counterstained with hematoxylin. Slide digital images were scanned with the NanoZoomer 2.0-HT slide scanner (Hamamatsu, Japan) at different magnifications.

Liu Y., Zhong X., Lin S., Xu H., Liang X., Wang Y., Xu J., Wang K., Guo X., Wang J., Yu M., Li C, & Xie C. (2022). Limosilactobacillus reuteri and caffeoylquinic acid synergistically promote adipose browning and ameliorate obesity-associated disorders. Microbiome, 10, 226.

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Quantifying Hepatic Steatosis and Inflammation

Cited 1 time

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Liver tissue was harvested at ZT 6 (12 p.m.) following fasting from ZT 0 (6 a.m.) on the final day of the experiment. Hepatocellular steatosis was measured by Oil Red O staining on 7 μm cryosections of liver tissue [30 (link)]. Images were scanned by bright-field light microscopy (Eclipse TE300, Nikon, NY, USA) at ×20 magnification. The area of Oil Red O lipid staining was measured using Image J software (Version 1.54g) [47 (link)]. Hepatic steatosis was further assessed by hematoxylin and eosin (H&E) staining of 5 μm liver tissue sections of paraffin-embedded liver tissue. Images were scanned by a Nanozoomer 2.0HT Slide Scanner (Hamamatsu Photonics, Bridgewater, NJ, USA), and the number and area of lipid droplets (stained negative for H&E) were measured using Image J. Hepatic inflammation was assessed by counting infiltrating immune cells in the H&E-stained sections. Inflammation scoring was performed by a trained pathologist blinded to the sample identities. Liver fibrosis was assessed by Sirius Red staining and quantified by Image J.

Das M., Kumar D., Sauceda C., Oberg A., Ellies L.G., Zeng L., Jih L.J., Newton I.G, & Webster N.J. (2024). Time-Restricted Feeding Attenuates Metabolic Dysfunction-Associated Steatohepatitis and Hepatocellular Carcinoma in Obese Male Mice. Cancers, 16(8), 1513.

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Immunohistochemistry and Toluidine Blue Staining of Optic Nerves

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WT spleens were fixed in 4% paraformaldehyde (PFA) in 0.1M sodium phosphate buffer (pH 7.4) overnight. Following transcardial perfusion with 4% PFA, optic nerves/chiasms were dissected and post-fixed in 4% PFA. Slides were deparaffinized in xylene, subjected to antigen retrieval, and incubated overnight with the appropriate antibodies (Supplementary Table S1), followed by incubation with biotinylated secondary antibodies (1:200; Vector Laboratories, Burlingame, CA). Signals were amplified with the Vectastain ABC kit (PK4000, Vector Laboratories), developed using the Vectastain DAB kit (SK4100, Vector laboratories), and counterstained with hematoxylin. Specimens were scanned using a NanoZoomer 2.0-HT slide scanner (Hamamatsu Photonics K.K., Japan) with a 20x objective, and the percentage of immunopositive cells calculated by manual counting. Optic nerve sections were deparaffinized, hydrated and stained in toluidine blue (1% toluidine blue in 70% ethanol; pH 2.3) for 3 minutes, washed in distilled water, dehydrated, and mounted. Stained optic nerves were imaged for quantification on a Leica ICC50W microscope using LAS EZ software.

de Andrade Costa A., Chatterjee J., Cobb O., Cordell E., Chao A., Schaeffer S., Goldstein A., Dahiya S, & Gutmann D.H. (2021). Immune deconvolution and temporal mapping identifies stromal targets and developmental intervals for abrogating murine low-grade optic glioma formation. Neuro-oncology Advances, 4(1), vdab194.

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Immunohistochemical Analysis of SGLT2 Protein

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The kidney slices were dried for
20 min using a cold blower and fixed with cold acetone for 10 min.
After rinsing in PBS, the endogenous peroxidase was blocked with 0.015%
H₂O₂ in PBS for 30 min. After rinsing in PBS,
the samples were incubated with the primary SGLT2 antibody (polyclonal,
raised in rabbit against amino acids 591–609 (ESAMEMNEPQAPAPSLFRQ-C)
of the SGLT2 protein,^{29 (link)} a gift from H Koepsell)
(dilution 1:250 in PBS/1% BSA) for 60 min at room temperature. After
rinsing in PBS, they were incubated with the secondary and third antibodies
(horseradish peroxidase-labeled goat–antirabbit and rabbit–antigoat,
respectively, from Dakopatts, Glostrup, Denmark, dilution 1:100 in
PBS/1% BSA and 1% AB serum), both for 30 min at room temperature.
After rinsing in PBS, the chromogenic reaction was visualized by incubation
with 3-amino-9-ethylcarbazole Substrate Kit (Life Technologies) for
10 min. After rinsing in demineralized water, the slices were counterstained
with hematoxyline (Fluka Chemie, Buchs, Switzerland), mounted in Kaiser’s
glycerol gelatin, and coverslipped. The IHC slices were scanned using
a Hamamatsu NanoZoomer 2.0 HT slide scanner (Hamamatsu Photonics,
Hamamatsu, Japan). Detailed images for this manuscript were created
using NDPviewer 2.

van der Hoek S., Antunes I.F., Attia K.A., Jacquet O., Heeres A., Bulthuis M., Zijlma R., Boersma H.H., van Goor H., Visser T.J., Heerspink H.J., Elsinga P.H, & Stevens J. (2021). GMP Compliant Synthesis of [18F]Canagliflozin, a Novel PET Tracer for the Sodium–Glucose Cotransporter 2. Journal of Medicinal Chemistry, 64(22), 16641-16649.

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Evaluating DACH1 Expression in HCC

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Commercially available tissue microarray (TMA) slides (BC03119, LV1201, US Biomax, Inc., Rockville, MD) containing histologically confirmed tissues from a variety of diseases were purchased for immunohistochemistry (IHC) analysis. Another tissue microarray (HLiv-HCC180Sur-02, Shanghai Biochip Co., Ltd., Shanghai, China) with 90 matched pairs of primary HCC samples and adjacent liver tissues was applied to evaluate the prognostic value of DACH1 based on its detailed survival data. Specific primary antibodies against DACH1 (ProteinTech Group) and cyclin D1 (Cell Signaling Technology) were used for IHC with a 2-step protocol. Whole slide image capture was performed on a Nano Zoomer 2.0 HT slide scanner (Ha mamatsu Photonics K.K., Hamamatsu, Japan) and captured by Nano Zoomer Digital Pathology view version 1.6.

Liu Y., Zhou R., Yuan X., Han N., Zhou S., Xu H., Guo M., Yu S., Zhang C., Yin T, & Wu K. (2015). DACH1 is a novel predictive and prognostic biomarker in hepatocellular carcinoma as a negative regulator of Wnt/β-catenin signaling. Oncotarget, 6(11), 8621-8634.

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Quantitative Analysis of Tumor Invasion

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The tissue sections stained with vimentin and CD56 were scanned using the whole slide scanner (NanoZoomer 2.0-HT slide scanner, Hamamatsu). The area of the spheroids in vitro and the tumors in vivo were measured using the program NanoZoomer Digital Pathology Version 2.3.11 from Hamamatsu. Using an area tool the spheroid or bulk tumor area without invasion were outlined and measured. The invasion area was measured as being the tumor cell area found outside the spheroids or tumor bulk using the software Visiomorph by making a classifier identifying the area of positive vimentin and CD56 staining subtracting the area of the spheroid or tumor bulk. The longest invasion distance was found using a linear measurement tool, measuring the perpendicular distance from the border of the spheroid or tumor bulk to the invasive front of the tumors.

Jensen S.S., Meyer M., Petterson S.A., Halle B., Rosager A.M., Aaberg-Jessen C., Thomassen M., Burton M., Kruse T.A, & Kristensen B.W. (2016). Establishment and Characterization of a Tumor Stem Cell-Based Glioblastoma Invasion Model. PLoS ONE, 11(7), e0159746.

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Immunohistochemical Analysis of HAUS6 and Ki-67

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Tissues were processed using routine immunohistochemistry methods as in our previous study (Shen et al., 2019 (link)). Tissue specimens collected in our laboratory were fixed, sectioned and stained with primary antibodies against HAUS6 and Ki-67 then secondary biotinylated antibody (Supplementary Table S6), followed by incubation with horseradish peroxidase-conjugated streptavidin (Maixing, Fuzhou, China). Background staining was assessed by omitting the primary antibody. Five randomly selected fields were examined using a Leica microscope (Wetzlar, Germany) at ×40 magnification.
For tissue arrays, slides were incubated with primary antibody against HAUS6 (dilution 1:500, Abcam, United Kingdom) as described (Wang et al., 2014 (link); Shen et al., 2019 (link)). Images were captured using a Nano Zoomer 2.0 HT slide scanner (Hamamatsu Photonics, Hamamatsu, Japan) and processed using Nano Zoomer Digital Pathology View 1.6 software. Staining intensity and the percentage of positively stained cells were determined as described (Huang et al., 2016 (link); Shen et al., 2019 (link)). Protein expression was calculated by multiplying the intensity and percentage scores together, giving expression scores ranging from 0 to 12.

Shen A., Liu L., Huang Y., Shen Z., Wu M., Chen X., Wu X., Lin X., Chen Y., Li L., Cheng Y., Chu J., Sferra T.J., Wei L., Zhuang Q, & Peng J. (2022). Down-Regulating HAUS6 Suppresses Cell Proliferation by Activating the p53/p21 Pathway in Colorectal Cancer. Frontiers in Cell and Developmental Biology, 9, 772077.

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Senescence β-Galactosidase Staining Assay

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Senescence assays were performed using the Senescence b‐Galactosidase Staining Kit (Cell Signaling) according to the manufacturer's protocol. Images were acquired using a Hamamatsu Nanozoomer 2.0HT Slide Scanner and quantified in independent images of 0.1 mm² covering the areas of interest using Keyence software.

Rocha L.R., Nguyen Huu V.A., Palomino La Torre C., Xu Q., Jabari M., Krawczyk M., Weinreb R.N, & Skowronska‐Krawczyk D. (2019). Early removal of senescent cells protects retinal ganglion cells loss in experimental ocular hypertension. Aging Cell, 19(2), e13089.

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Histological Analysis of Postnatal Mouse Brains

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For histology, brains were harvested from postnatal day 14 (P14) mice and fixed in Bouin’s fixative (Electron Microscopy Sciences) overnight (ON), dehydrated in ethanol, paraffin-embedded, and coronal sections (20 μm) obtained, before being stained with hematoxylin and mounted with Entellan (Millipore). Brain sections were examined using Leica MZ-16 stereomicroscope using the NanoZoomer 2.0-HT slide scanner and analyzed with the Hamamatsu NDP viewer software (Hamamatsu).

Dos-Santos Carvalho S., Moreau M.M., Hien Y.E., Garcia M., Aubailly N., Henderson D.J., Studer V., Sans N., Thoumine O, & Montcouquiol M. (2020). Vangl2 acts at the interface between actin and N-cadherin to modulate mammalian neuronal outgrowth. eLife, 9, e51822.

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Depth of Invasion in Head and Neck Cancer

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The DOI was measured for all surgical specimens based on the hematoxylin and eosin slide. The DOI was defined and measured as a plumb-line from the basal membrane of the closest normal adjacent mucosa to the deepest point of invasion, in line with the recommendation from the 8^th edition of the AJCC (18 (link)).
All hematoxylin and eosin slides were collected from the Department of Pathology of the Erasmus University Medical Center and scanned by the NanoZoomer 2.0-HT slide scanner (Hamamatsu Photonics, Hamamatsu, Japan). Slides were reviewed by a head and neck pathologist (SK) using the NanoZoomer digital pathology (NDP) viewer 2.5.19 (Hamamatsu Photonics, Hamamatsu, Japan).
The patients were divided based on DOI into a group with DOI ≤4 mm and a group with DOI >4 mm, based on the DOI cut-off value >4 mm used at our institute.

Aaboubout Y., van der Toom Q.M., de Ridder M.A., De Herdt M.J., van der Steen B., van Lanschot C.G., Barroso E.M., Nunes Soares M.R., ten Hove I., Mast H., Smits R.W., Sewnaik A., Monserez D.A., Keereweer S., Caspers P.J., Baatenburg de Jong R.J., Bakker Schut T.C., Puppels G.J., Hardillo J.A, & Koljenović S. (2021). Is the Depth of Invasion a Marker for Elective Neck Dissection in Early Oral Squamous Cell Carcinoma?. Frontiers in Oncology, 11, 628320.

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