Pannoramic 250 flash 3

Manufactured by 3DHISTECH

Sourced in Hungary

The Pannoramic 250 Flash III is a high-speed digital slide scanner designed for microscopic imaging. It is capable of scanning glass slides at a high resolution and capturing digital images for further analysis and archiving.

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Lab products found in correlation

Caseviewer, by 3DHISTECH (12 mentions) Caseviewer 2, by 3DHISTECH (7 mentions) Pannoramic viewer software, by 3DHISTECH (5 mentions) Lsm 780 confocal microscope, by Zeiss (3 mentions) Ab16667, by Abcam (3 mentions) Antigen unmasking solution, by Vector Laboratories (2 mentions) Antigen unmarking solution, by Vector Laboratories (2 mentions) Technovit 7200 vlc, by Kulzer (2 mentions) Caseviewer software, by 3DHISTECH (2 mentions) Dm5500, by Leica (2 mentions) Blocking buffer, by Thermo Fisher Scientific (1 mentions) Haematoxylin, by Avantor (1 mentions) Keratin 5, by BioLegend (1 mentions) Cytokeratin 8, by BioLegend (1 mentions) Il 6r, by R&D Systems (1 mentions) Tunel assay kit, by Abcam (1 mentions) Nb100 2076, by Novus Biologicals (1 mentions) Alexa fluor 488 conjugated goat anti mouse secondary antibody, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 conjugated goat anti rabbit secondary antibody, by Thermo Fisher Scientific (1 mentions) Nb600 408, by Novus Biologicals (1 mentions)

Close spelling variants of this product

Pannoramic 250 (67 citations) Pannoramic 250 FLASH (54 citations) Pannoramic 250 Flash III scanner (15 citations) Pannoramic 250 Flash III slide scanner (14 citations) Pannoramic 250 Flash III whole-slide scanner (4 citations) PANNORAMIC® 250 Flash III DX (3 citations) Pannoramic 250 Flash III digital scanner (2 citations) Pannoramic 250 Flash III digital (2 citations)

82 protocols using pannoramic 250 flash 3

Standardized FFPE Tumor Sample Preparation

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All diagnostic slides, from the lumpectomy surgical sample, for each patient were pathologist-reviewed (IMM and MST), and the best representative (to ensure the presence of adequate tumor tissue for analysis, morphological variation, and to confirm the pure DCIS diagnosis) formalin-fixed paraffin-embedded (FFPE) tumor blocks (donor) for each patient’s specimen were retrieved and included in the study. A fresh full-face section of 4 μm thickness was cut from each selected block, stained with H&E to standardize the consistency of staining quality, and again pathologist-reviewed (IMM and MST). Slide scanning was performed with a slide scanner using a × 40 magnification objective lens (0.24 μm/pixel) (Pannoramic 250 Flash III, 3DHISTECH) (Additional file 1: Supplementary methodology). Images were viewed at a maximum of × 400 magnification using a built-in functionality of image processing software (ImageScope, ver. 12.3.2.8013, Leica Microsystems). The slides were reviewed for image quality, those with out-of-focus areas re-scanned, and those with folded over tissues removed from the analysis.

Klimov S., Miligy I.M., Gertych A., Jiang Y., Toss M.S., Rida P., Ellis I.O., Green A., Krishnamurti U., Rakha E.A, & Aneja R. (2019). A whole slide image-based machine learning approach to predict ductal carcinoma in situ (DCIS) recurrence risk. Breast Cancer Research : BCR, 21, 83.

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Immunofluorescence Analysis of Pdcd4 in Mice

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Mice were anesthetized by 5% chloral hydrate (7.5 ml/kg, i.p.), then, they were transcardially perfused with saline and 4% PFA. The brains were harvested, embedded in OCT, and sectioned 40 μm thick. The slides were washed by PBS for twice to remove OCT. For immunofluorescence, slides were incubated in 0.4% TritonX-100- diluted donkey serum to 10% which was used to block nonspecific staining for 30 min. Primary antibodies: anti-Pdcd4 (1:100), anti-NuN (1:500), anti-GFAP (1:500) were used to incubated with slides overnight at 4 °C, after that, they were washed with PBS three times and secondary antibody (1:500) incubated the slides at room temperature for 1 h. Washing in PBS three times, slides were mounted with cover glass, n = 4 per group. All of the images were captured with a Zeiss LSM780 confocal microscope (Oberkochen, Germany) at Microstructural Platform of Shandong University. Images were analyzed by NIH Image J. For Nissl staining, sections were cut at 40 μm intervals, and according to the standard procedure staining was done. Images were acquired with a light microscope Pannoramic 250 Flash III (3D Histech, Budapest, HUNGARY).

Li Y., Jia Y., Wang D., Zhuang X., Li Y., Guo C., Chu H., Zhu F., Wang J., Wang X., Wang Q., Zhao W., Shi Y., Chen W, & Zhang L. (2020). Programmed cell death 4 as an endogenous suppressor of BDNF translation is involved in stress-induced depression. Molecular Psychiatry, 26(6), 2316-2333.

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Immunohistochemistry and Immunofluorescence Protocols

Cited 1 time

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Immunohistochemistry was performed using standard protocol as we previously described (Chen et al., 2017a (link); Zhao et al., 2017 (link)). In brief, a pressure cooker (95 °C for 30 min followed by 120 °C for 10 s) was used for antigen retrieval using antigen unmasking solution (Vector Laboratories). Antibodies specific to Mac-2 (BioLegend, #125403), S473P-AKT, PTEN, YAP1, LOX, β1 integrin, CD68 (Abcam, #ab53444), cleaved caspase 3 (CST, #9661S), Ki67 (Thermo Scientific, #RM-9106-S1), CD31 (Abcam, #ab28364) were used in this study. The human and mouse tumor tissue sections were reviewed and scored as reported recently (Chen et al., 2017a (link)). Slides were scanned using Pannoramic 250 Flash III (3DHISTECH Ltd) and images were captured through Pannoramic Viewer software (3DHISTECH Ltd). The studies related to human specimens were approved by the MD Anderson Institutional Review Board under protocol #PA14–0420.
Immunofluorescence was performed using the standard protocol in cells and tissues. Antibodies specific to F4/80 (Abcam, #ab6640), CD68, CD206 (R&D system, #AF2535), LOX, YAP1, β1 integrin, CD31, cleaved caspase 3 and SPP1 (Santa Cruz, #sc-21742) were used. Images were captured using a fluorescence microscope (Leica DMi8).

Chen P., Zhao D., Li J., Liang X., Li J., Chang A., Henry V.K., Lan Z., Spring D.J., Rao G., Wang Y.A, & DePinho R.A. (2019). Symbiotic macrophage-glioma cell interactions reveal synthetic lethality in PTEN null glioma. Cancer cell, 35(6), 868-884.e6.

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Immunohistochemical Staining of IGF2BP1

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For immunohistochemical staining of IGF2BP1 protein, paraffin-embedded human colon-tumor tissues were sectioned and de-paraffinized with xylene followed by dehydration with a graded series of alcohol. Antigen retrieval was conducted by heating in 0.1 M citrate buffer, followed by blocking with blocking buffer (Thermo Fisher Scientific, Waltham, MA, USA). Slices were stained with anti-IGF2BP1 antibody (1:100) at 4°C overnight. Then, samples on slices were incubated with a horseradish peroxidase (HRP)-labeled goat anti-rabbit secondary antibody. After washing, sections were scanned by Pannoramic 250 flash III (3DHistech, Hungary).

Zhang X.L., Li K.J., Feng J.X., Liu G.J, & Feng Y.L. (2021). Blocking the IGF2BP1-promoted glucose metabolism of colon cancer cells via direct de-stabilizing mRNA of the LDHA enhances anticancer effects. Molecular Therapy. Nucleic Acids, 23, 835-846.

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Histological Analysis of Bone Samples

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Bones were fixed in neutral buffered formalin for 48 h after which they were transferred to 70% ethanol. Bones were then decalcified in 10% EDTA, embedded in paraffin and 5 µm sections produced. Sections were dewaxed in xylene, rehydrated through graded alcohols and stained in haematoxylin (VWR) and eosin. The sections were dehydrated through graded alcohols and coverslips were mounted in DPX and imaged using an Olympus BX51 (Olympus Life Sciences) and Pannoramic 250 Flash III slide scanner (3DHISTECH).

Green D., Singh A., Tippett V.L., Tattersall L., Shah K.M., Siachisumo C., Ward N.J., Thomas P., Carter S., Jeys L., Sumathi V., McNamara I., Elliott D.J., Gartland A., Dalmay T, & Fraser W.D. (2023). YBX1-interacting small RNAs and RUNX2 can be blocked in primary bone cancer using CADD522. Journal of Bone Oncology, 39, 100474.

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Histological Analysis of Circular Defects

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The specimens were decalcified with a 10% ethylenediaminetetraacetic acid solution for 2 weeks before they were dehydrated by a series of ethanol solutions of increasing concentrations and embedded in paraffin. A coronal section with a thickness of 5 μm through the center of the circular defect was obtained and stained with hematoxylin and eosin. The prepared specimens were examined by light microscopy. After a microscopic examination, a photograph of each slide was taken using a digital slide scanner (PANNORAMIC 250 Flash III, 3DHISTECH, Budapest, Hungary), and the resulting images were saved on a computer for analysis. A single blinded, calibrated examiner examined all of the images 3 times using the same software (ImageJ 1.53e, National Institutes of Health, Bethesda, MD, USA). The following parameters were recorded for each defect site:

Li L., Lee J., Cho Y.D., Kim S., Seol Y.J., Lee Y.M, & Koo K.T. (2022). The optimal dosage of hyaluronic acid for bone regeneration in rat calvarial defects. Journal of Periodontal & Implant Science, 53(4), 259-268.

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Comprehensive Immunohistochemistry Profiling

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Immunohistochemistry was performed as previously described (17 (link)). A pressure cooker (95°C for 30 min followed by 120°C for 10 s) was used for antigen retrieval using Antigen Unmarking Solution (Vector Laboratories). Antibodies specific to CHD1 (Sigma, #HPA022236), AKT-473P (Cell Signaling, #4060s), Keratin 5 (BioLegend 905501), Cytokeratin-8 (BioLegend 904801), CD8 (Bioss bs-0648R), Ly6G (BioLegend 127601), CD15 (DAKO M3631), IL‑6R (R&D Systems AF1830-SP) and Ki67 (Thermo Scientific RM 9106-S1) were purchased. TUNEL staining was performed using the TUNEL Assay Kit (Abcam ab206386) according to manufacturer’s instructions. Slides were scanned using Pannoramic 250 Flash III (3DHISTECH Ltd) and images were captured through Pannoramic Viewer software (3DHISTECH Ltd). Human prostate hyperplasia and cancer tissue array samples were purchased from US Biomax (PR753a).

Zhao D., Cai L., Lu X., Liang X., Li J., Chen P., Ittmann M., Shang X., Jiang S., Li H., Meng C., Flores I., Song J.H., Horner J.W., Lan Z., Wu C.J., Li J., Chang Q., Chen K.C., Wang G., Deng P., Spring D.J., Wang Y.A, & DePinho R.A. (2020). Chromatin Regulator, CHD1, Remodels the Immunosuppressive Tumor Microenvironment in PTEN-Deficient Prostate Cancer. Cancer discovery, 10(9), 1374-1387.

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Histological Analysis of Lung Tissue

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For general histology, in paraffin sections from lung tissue samples, Hematoxylin and eosin (H&E) staining were performed according to standard steps.
For immunofluorescence, 6-μm sections were treated with xylene and graded alcohols. Antigen retrieval was accomplished by microwaving slides in EDTA antigen retrieval buffer (pH 8.0). The slides were blocked with 5% BSA for 1 h and then incubated overnight at 4°C with rabbit polyclonal anti-collagen-I antibody (1:50; NB600-408; NOVUS), mouse monoclonal anti-elastin antibody (1:50; NB100-2076; NOVUS), or mouse anti-Fibrinogen beta chain (FGB) antibody (1:50; M01204-1; Boster), followed by Alexa Fluor 488-conjugated goat anti-rabbit secondary antibodies (A-11008, Invitrogen) or Alexa Fluor 488-conjugated goat anti-mouse secondary antibodies (A32723, Invitrogen) for 1h at room temperature and nuclei were stained with DAPI (62248, Invitrogen). The Pannoramic 250 Flash III (3DHISTECH) was used to scan the sections, and CaseViewer 2.4 was used to record the images.

Gong H., Chen L., He Y., Hua K., Ma B., Gao Y., Xu X., Hu X, & Jin H. (2022). Pleural thickening induced by Glaesserella parasuis infection was linked to increased collagen and elastin. Frontiers in Cellular and Infection Microbiology, 12, 952377.

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SERPINB5 and KRT19 Expression Analysis in FFPE Tissue

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Formalin-fixed paraffin-embedded tissue was sectioned (4μm) and analyzed for SERPINB5 and Cytokeratin-19 (KRT19) expression using ViewRNA™ ISH Tissue Assay Kit (ThermoFisher), with probes against SERPINB5 (VA1–12247-VT) and cytokeratin-19 (VA6–10947-VT) (ThermoFisher), according to manufacturer’s protocol. In short: tissue was deparaffinized in Xylene and 100% ethanol. The sections were pretreated for 10min at 90–95°C, followed by protease digestion for 10min at 40°C and then fixed in 10% normal buffered formalin followed by hybridization with the target probes against SERPINB5 and KRT19 for 2h in 40°C. The sections were stored overnight in storage buffer before proceeding with signal amplification and detection. The sections were preamplified for 25min at 40°C followed by amplifier hybridization for 15min at 40°C. Followed by incubation with label probes at 40°C for 15min and addition of substrate. FastRed label probe and substrate incubation was performed first. Sections were then counterstained in Gil’s hematoxylin, dipped in 0.01% ammonium hydroxide followed by DAPI staining (1μg/ml for 10min). Slides were mounted with Prolong gold antifade mountant with DAPI (Thermofisher). The confocal images were taken on a Zeiss LSM 710 microscope and the IHC images were scanned on the 3DHISTECH Pannoramic 250 flash III.

Tian C., Öhlund D., Rickelt S., Lidström T., Huang Y., Hao L., Zhao R.T., Franklin O., Bhatia S.N., Tuveson D.A, & Hynes R.O. (2020). Cancer-cell-derived matrisome proteins promote metastasis in pancreatic ductal adenocarcinoma. Cancer research, 80(7), 1461-1474.

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Fetal Liver Cell Cytospin Staining

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Up to 50,000 cultured fetal liver cells were suspended in 150μl PBS and immobilized on twin frosted glass microscope slides (Fisher Scientific) by cytospin at 200rpm, low acceleration for 5 minutes in a Shandon Cytospin3 (Thermo Scientific). Air-dried slides were submerged in May-Grünwald Eosin methylene blue Q Path stain (VWR) for 3 minutes, rinsed in tap water and submerged in 5% Giemsa’s stain (VWR) for 20 minutes. Slides were subsequently air dried, mounted and scanned using the Pannoramic 250 Flash III (3DHISTECH). Images were acquired with the Pannoramic 250 software and analyzed with the Pannoramic Viewer software (3DHISTECH).

Draper J.E., Sroczynska P., Fadlullah M.Z., Patel R., Newton G., Breitwieser W., Kouskoff V, & Lacaud G. (2018). A novel prospective isolation of murine fetal liver progenitors to study in utero hematopoietic defects. PLoS Genetics, 14(1), e1007127.

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