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Pannoramic 250 digital scanner

Manufactured by 3DHISTECH
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Sourced in Hungary

The Pannoramic 250 is a digital slide scanner designed for high-throughput scanning of microscope slides. It captures digital images of entire glass slides at high resolution, enabling efficient digitization of pathology samples.

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Lab products found in correlation

Autostainer link, by Agilent Technologies (2 mentions) Caseviewer, by 3DHISTECH (2 mentions) Biotinylated anti mouse antibody, by Vector Laboratories (2 mentions) Bx61 microscope, by 3DHISTECH (2 mentions) Anti iba1, by Fujifilm (2 mentions) Vectastain abc kit, by Vector Laboratories (2 mentions) Anti mouse alexa594, by Thermo Fisher Scientific (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) 8 μm pore polycarbonate filter, by Corning (1 mentions) Matrigel, by Corning (1 mentions) Transwell cell culture insert, by Corning (1 mentions) Eclipse ci microscope, by Nikon (1 mentions)

8 protocols using pannoramic 250 digital scanner

1

Histological Analysis of Tumor Samples

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Tumours were dissected and fixed in 10% buffered formaldehyde, and were then embedded in paraffin wax. Sections were cut at a 3-µm thickness, mounted on slides, and stained in eosin and haematoxylin. The slides were scanned using the Pannoramic 250 Digital Scanner (3D HISTECH).
He J., Zhou Y., Arredondo Carrera H.M., Sprules A., Neagu R., Zarkesh S.A., Eaton C., Luo J., Gartland A, & Wang N. (2020). Inhibiting the P2X4 Receptor Suppresses Prostate Cancer Growth In Vitro and In Vivo, Suggesting a Potential Clinical Target. Cells, 9(11), 2511.
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2

Automated Immunohistochemistry Staining Protocol

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Immunohistochemistry (IHC) was done in automated system using the Dako Autostainer Link. Formalin fixed, paraffin sections were cut at 4 μm and rehydrated to water. Heat induced epitope retrieval was performed with FLEX TRS High pH Retrieval buffer for 20 min. After peroxidase blocking, the specific monoclonal antibodies were applied at room temperature for 20 min. The FLEX + Rabbit EnVision System was used for detection. DAB chromogen was then applied for 10 min. Slides were counterstained with Mayers hematoxylin for 5 s and then dehydrated and coverslipped. Slides were scanned on a Pannoramic 250 digital scanner (3D HISTECH Ltd., Budapest, Hungary) and images scored using the software program ‘Case viewer’ (3D HISTECH Ltd., Budapest, Hungary). Negative controls were included in each staining run.
Hadgu E., Seifu D., Tigneh W., Bokretsion Y., Bekele A., Abebe M., Sollie T., Merajver S.D., Karlsson C, & Karlsson M.G. (2018). Breast cancer in Ethiopia: evidence for geographic difference in the distribution of molecular subtypes in Africa. BMC Women's Health, 18, 40.
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3

Transwell Assay for Cell Migration

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As C4-2B4 monolayers were easily detached with physical manipulation, these cells were not suitable for the examination of migration using scratch assays. A Transwell method was therefore adopted to test the migration of C4-2B4 cells. C4-2B4 cells were seeded at 2 × 105 cells per well in 200 μL serum-free medium containing 1 μM 5-BDBD, 1.5 μM PSB-12062, or vehicle on top of the Transwell® cell culture inserts (6.5 mm with an 8-μm pore polycarbonate filter, Corning, New York, NY, USA), while 500 μL of 10% FBS DMEM was added to the lower chamber as the chemoattractant. After 16 h of incubation, the inserts were fixed in 100% ethanol for 5 min, stained in eosin for 1 min and haematoxylin for 5 min, air-dried and mounted on slides in Faramount. Finally, the slides were scanned using the Pannoramic 250 Digital Scanner (3D HISTECH, Budapest, Hungary), and the percentage areas covered by migrated cells were measured by using ImageJ software.
He J., Zhou Y., Arredondo Carrera H.M., Sprules A., Neagu R., Zarkesh S.A., Eaton C., Luo J., Gartland A, & Wang N. (2020). Inhibiting the P2X4 Receptor Suppresses Prostate Cancer Growth In Vitro and In Vivo, Suggesting a Potential Clinical Target. Cells, 9(11), 2511.
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4

Quantifying Amyloid and Microglial Responses

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30 μm thick sagittal free-floating sections were incubated with anti-Iba1 (1:500, cat#019-19741, Wako, Richmond, VA, USA), 6E10 (1:1000, cat#9320-500, Covance, Princeton, NJ, USA), or C1q (1:1000, cat#ab182451, Abcam, Cambridge, MA, USA) antibodies. Sections probed with Iba1 and 6E10 antibodies were incubated with anti-rabbit Alexa 488 (1:400, cat#A-11008, Thermo Fisher Scientific, Grand Island, NY, USA) and anti-mouse Alexa 594 (1:400, cat#A-11005, Thermo Fisher Scientific) secondary antibodies. Sections probed with C1q were incubated with biotinylated anti-mouse antibody (1:1000, BA-9200, Vector laboratories, Burlingame, CA, USA) and developed with Vectastain ABC Kit (PK-4000, Vector Laboratories). (see suppl. methods for details). Images were acquired using an Olympus BX61 microscope or a Pannoramic 250 digital scanner (3DHISTECH, Budapest, Hungary).
For measuring percentage of 6E10 immunoreactive areas, 6E10-immunolabeled sections were thresholded and the percentage of 6E10 positive area was calculated using Fiji (v2.0.0) and the “measure” function. For measuring microglial number, Iba1 positive cells in the region of interest (ROI) were manually counted and number was normalized to the area of the ROI. ROI (prefrontal cortex, hippocampus) were determined and outlined by manual tracing.
Haure-Mirande J.V., Wang M., Audrain M., Fanutza T., Kim S.H., Heja S., Readhead B., Dudley J.T., Blitzer R.D., Schadt E.E., Zhang B., Gandy S, & Ehrlich M.E. (2018). Integrative approach to sporadic Alzheimer’s disease: deficiency of TYROBP in cerebral Aβ amyloidosis mouse normalizes clinical phenotype and complement subnetwork molecular pathology without reducing Aβ burden. Molecular Psychiatry, 24(3), 431-446.
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5

Immunohistochemical Analysis of Brain Inflammation

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30µm thick  sagittal free-floating sections were incubated with anti-Iba1 (1:500, cat#019-19741, Wako, Richmond, VA, USA), 6E10 (1:1000, cat#9320-500, Covance, Princeton, NJ, USA), or C1q (1:1000, cat#ab182451, Abcam, Cambridge, MA, USA) antibodies. Sections probed with Iba1 and 6E10 antibodies were incubated with anti-rabbit Alexa 488 (1:400, cat#A-11008, Thermo Fisher Scientific, Grand Island, NY, USA) and anti-mouse Alexa 594 (1:400, cat#A-11005, Thermo Fisher Scientific) secondary antibodies. Sections probed with C1q were incubated with biotinylated anti-mouse antibody (1:1000, BA-9200, Vector laboratories, Burlingame, CA, USA) and developed with Vectastain ABC Kit (PK-4000, Vector Laboratories) (see Suppl. methods for details). Images were acquired using an Olympus BX61 microscope or a Pannoramic 250 digital scanner (3DHISTECH, Budapest, Hungary).
For measuring percentage of 6E10-immunoreactive areas, 6E10-immunolabeled sections were thresholded and the percentage of 6E10 positive area was calculated using Fiji (v2.0.0) and the “measure” function. For measuring microglial number, Iba1 positive cells in the region of interest (ROI) were manually counted and number was normalized to the area of the ROI. ROI (PFC and hippocampus) were determined and outlined by manual tracing.
Haure-Mirande J.V., Wang M., Audrain M., Fanutza T., Kim S.H., Heja S., Readhead B., Dudley J.T., Blitzer R.D., Schadt E.E., Zhang B., Gandy S, & Ehrlich M.E. (2018). Integrative approach to sporadic Alzheimer’s disease: deficiency of TYROBP in cerebral Aβ amyloidosis mouse normalizes clinical phenotype and complement subnetwork molecular pathology without reducing Aβ burden. Molecular Psychiatry, 24(3), 431-446.
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6

Transwell Invasion Assay for PC3 and C4-2B4 Cells

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Transwell® cell culture inserts (6.5 mm with an 8-μm pore polycarbonate filter, Corning) were first coated with 0.5 μg/mL Matrigel (Corning) in serum-free medium at 37 °C for 2 h. 5 × 104 PC3 or C4-2B4 cells were then seeded into the top chamber with 200 μL serum-free medium containing antagonists (1.0 μM 5-BDBD or 1.5 μM PSB-12062) or vehicles, and were treated with 5 μg/mL Mitomycin C for 3 h. 500 μL of 10% FBS DMEM was added to the lower chamber as the chemoattractant. After 72 h of incubation, the inserts were fixated in 100% ethanol for 5 min, stained in eosin for 1 min and haematoxylin for 5 min, and then mounted on a slide. The slides were scanned using the Pannoramic 250 Digital Scanner (3D HISTECH) at a 1.2× magnification. The percentage areas covered by invaded cells were analysed by using ImageJ software.
He J., Zhou Y., Arredondo Carrera H.M., Sprules A., Neagu R., Zarkesh S.A., Eaton C., Luo J., Gartland A, & Wang N. (2020). Inhibiting the P2X4 Receptor Suppresses Prostate Cancer Growth In Vitro and In Vivo, Suggesting a Potential Clinical Target. Cells, 9(11), 2511.
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7

Automated IHC Staining for p16

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Immunohistochemistry (IHC) for p16 was done using an automated system, the Dako Autostainer Link. Formalin-fixed paraffin sections were cut at 4 microns and rehydrated with water. Heat-induced epitope retrieval was performed with the FLEX TRS High-pH Retrieval Buffer for 20 minutes. After peroxidase blocking, the specific monoclonal antibody (source and dilution: p16 clone G175-405 from BD, USA and diluted 1/25) was applied at room temperature for 20 minutes. The FLEX + Rabbit EnVision System was used for detection. DAB chromogen was then applied for 10 minutes. Slides were counterstained with Mayers hematoxylin for 5 seconds and then dehydrated and coverslipped. Slides were scanned on a Pannoramic 250 digital scanner (3D HISTECH Ltd., Budapest, Hungary) and images scored using the software program “CaseViewer” (3D HISTECH Ltd., Budapest, Hungary). Negative controls were included in the run.
Gebregzabher E., Seifu D., Tigneh W., Bokretsion Y., Bekele A., Abebe M., Lillsunde-Larsson G., Karlsson C, & Karlsson M.G. (2021). Detection of High- and Low-Risk HPV DNA in Archived Breast Carcinoma Tissues from Ethiopian Women. International Journal of Breast Cancer, 2021, 2140151.
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8

Comprehensive Histological Analysis of Tumor Samples

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After fixing with 4% paraformaldehyde, tumor, heart, liver, spleen, lung, kidney and brain fragments were embedded with paraffin and sectioned. H&E staining was then conducted according to the manufacturer’s protocol (Servicebio, Wuhan, China). Moreover, immunohistochemistry staining for caspase-3 and Ki-67 was performed on tumor sections to determine apoptosis and proliferation of tumor cells, respectively. Images were acquired using an Eclipse Ci microscope (Nikon Corporation, Tokyo, Japan) or a Pannoramic 250 digital scanner (3DHistech, Budapest, Hungary).
Chu C., Bao Z., Sun M., Wang X., Zhang H., Chen W., Sui Y., Li J., Zhuang Y, & Wang D. (2020). NIR Stimulus-Responsive PdPt Bimetallic Nanoparticles for Drug Delivery and Chemo-Photothermal Therapy. Pharmaceutics, 12(7), 675.
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