Histoquant

Manufactured by 3DHISTECH

Sourced in Hungary

Histoquant is a laboratory equipment product designed for quantitative analysis of histological samples. It provides automated digital image analysis capabilities to measure and quantify various parameters within biological specimens.

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Spelling variants of this product

HistoQuant software (12 citations) HistoQuant module (5 citations) Histoquant program (3 citations) HistoQuant quantification module (2 citations)

18 protocols using histoquant

Hippocampal Sclerosis Grading Protocol

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We determined ILAE [15] (link) and Wyler [16] semi-quantitative gradings of HS from resected specimens. ILAE ratings included specimens with severe neuronal cell loss and gliosis predominantly in CA1 and CA4 regions (ILAE type 1), predominantly in CA1 (ILAE type 2), predominantly in CA4 (ILAE type 3) or with no HS. Wyler grades provided grades of overall hippocampal cell loss ranging from no pathology to Grade 4, with three incremental grades (mild, moderate, moderate to marked, and marked). Furthermore, quantitative neuronal cell density (number of neurons/μm²) was obtained from each specimen, as recently described [17] (link). After scanning of Neu-N immunohistochemistry using a Mirax scanner (3DHistech, Hungary), rectangular regions-of-interest (ROIs) within hippocampal subfields CA1-4 were selected. NeuN-positive cells within these ROIs were counted with a quantitative software solution (HistoQuant, 3DHistech, Hungary) after manual control of correct cell identification.

Elkommos S., Weber B., Niehusmann P., Volmering E., Richardson M.P., Goh Y.Y., Marson A.G., Elger C, & Keller S.S. (2016). Hippocampal internal architecture and postoperative seizure outcome in temporal lobe epilepsy due to hippocampal sclerosis. Seizure, 35, 65-71.

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Automated Plaque Quantification in Histological Specimens

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Entire DAPI/Thioflavin S-stained sections were scanned using a 3D Scan Pannoramic Histech scanner (3D Histech Kft. Budapest, Hungary) at a resolution of 0.23 μm per pixel. Images were analyzed using Pannoramic Viewer 1.15.2 SP2 RTM (3D Histech kft.) or Histoquant (3D Histech kft.) software, which provides a detailed morphometric analysis with precise measurements of different histological parameters at high resolution (Figure 2). Plaques were detected automatically, and the number of plaques per section and the area of each plaque was evaluated.

Díaz González M., Buberman A., Morales M., Ferrer I, & Knafo S. (2021). Aberrant Synaptic PTEN in Symptomatic Alzheimer’s Patients May Link Synaptic Depression to Network Failure. Frontiers in Synaptic Neuroscience, 13, 683290.

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Quantifying Hepatic Collagen Fibers

Cited 1 time

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Paraffin-embedded liver sections (2–3 µm) were treated with 0.1% Sirius red in saturated picric acid (Chroma, Münster, Germany) to detect collagen fibers. Hepatic Sirius red stainings were digitalized using Pannoramic MIDI (3DHistech, Budapest, Hungary) and quantified using Histoquant (3DHistech, Budapest, Hungary) as previously described^{8 (link),35 (link)}.

Klein S., Kleine C.E., Pieper A., Granzow M., Gautsch S., Himmit M., Kahrmann K., Schierwagen R., Uschner F.E., Magdaleno F., Naoum M.E., Kristiansen G., Walther T., Bader M., Sauerbruch T, & Trebicka J. (2019). TGR(mREN2)27 rats develop non-alcoholic fatty liver disease-associated portal hypertension responsive to modulations of Janus-kinase 2 and Mas receptor. Scientific Reports, 9, 11598.

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Immunohistochemical Analysis of Liver Tissue

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Staining for JAK2, pJAK2, and α-SMA was performed in cryosections from liver tissue (3 and 7 μm). The detailed method is described in the Supporting Information. The amount of staining was evaluated by computational analysis (Histoquant; 3DHistech, Budapest, Hungary), as described previously.^{27 ,28}Quantification (% of stained area) of immunohisto-chemical (IHC) staining is expressed as mean ± SEM of all experiments. For representative sections, please see the Supporting Figures.

Granzow M., Schierwagen R., Klein S., Kowallick B., Huss S., Linhart M., Reza Mazar I.G., Görtzen J., Vogt A., Schildberg F.A., Gonzalez-Carmona M.A., Wojtalla A., Krämer B., Nattermann J., Siegmund S.V., Werner N., Fürst D.O., Laleman W., Knolle P., Shah V.H., Sauerbruch T, & Trebicka J. (2014). Angiotensin-II Type 1 Receptor-Mediated Janus Kinase 2 Activation Induces Liver Fibrosis. Hepatology (Baltimore, Md.), 60(1), 334-348.

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Quantifying Immune Infiltration in Breast Cancer

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Formalin-fixed and paraffin-embedded breast carcinoma sections were analyzed following double immunohistochemical staining for CAIX and CD8. Scanned digital slides were evaluated for both labelings, and CD8-related signal was determined within and outside CAIX-positive tumor areas using the HistoQuant (3DHistech) image analysis software. Statistical analysis was performed using GraphPad Prism software.

Juhász P., Hasulyó D., Bedekovics J., Beke L., Kacsala N., Török M, & Méhes G. (2023). Carbonic Anhydrase IX (CAIX) Expressing Hypoxic Micro-environment Hampers CD8+ Immune Cell Infiltrate in Breast Carcinoma. Applied immunohistochemistry & molecular morphology : AIMM, 31(1).

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Quantitative Immunohistochemistry of Tau Pathology

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Immunhistochemistry was performed on 4 μm thick paraffin sections of formalin-fixed tissue and on 10 μm thick brain sections of frozen brain tissue using standard techniques. The immunohistochemical tau-staining was performed semi-automatically on a BenchMark device (Ventana, now Hoffmann-LaRoche, Basel, Switzerland) with mouse monoclonal AT8 antibody raised against hyperphosphorylated tau (Ser202/Thr205, 1:200, Invitrogen/Thermofisher, Carlsbad, CA, USA) on adjacent sections of those used in the ARG. The immunostained sections were digitized at 20x magnification with a Mirax Midi scanner (Zeiss, Carl Zeiss MicroImaging GmbH, Jena, Germany). Five cortical gray matter regions of interest were drawn manually and the tau load (in %) was quantified using the Pannoramic Viewer (1.15.2) software with HistoQuant (3DHISTECH, Budapest, Hungary) based on color threshold values (see Figure 1). All brain sections included are shown in the Supplementary Material.

Willroider M., Roeber S., Horn A.K., Arzberger T., Scheifele M., Respondek G., Sabri O., Barthel H., Patt M., Mishchenko O., Schildan A., Mueller A., Koglin N., Stephens A., Levin J., Höglinger G.U., Bartenstein P., Herms J., Brendel M, & Beyer L. (2021). Superiority of Formalin-Fixed Paraffin-Embedded Brain Tissue for in vitro Assessment of Progressive Supranuclear Palsy Tau Pathology With [18F]PI-2620. Frontiers in Neurology, 12, 684523.

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Quantitative Evaluation of Lipid Droplets in Liver

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For quantitative evaluation of LDs in the liver, ORO-stained sections were analyzed with a slide scanner (Pannoramic Histech 3D Scan, 3D Histech kft., Budapest, Hungary). Quantitative studies were performed using the Pannoramic Viewer and Histoquant softwares (3D Histech kft.) as follows: 5 randomly areas of the liver tissue measuring 100,000 μm² per area were outlined in each section (n = 3 sections/animal), performing a total analysis of 1,500,000 μm² of tissue area per animal. Areas were randomly chosen in the liver tissue. In infected tissues, areas were not marked over inflammatory infiltrates or granulomas. In the demarcated areas, LD quantification within hepatocytes was performed by counting all LDs and all nuclei of hepatocytes, considering one nucleus/cell. The number of LDs per hepatocyte was obtained by dividing the total number of LDs per the total number of nuclei of hepatocytes.

Amaral K.B., Silva T.P., Malta K.K., Carmo L.A., Dias F.F., Almeida M.R., Andrade G.F., Martins J.S., Pinho R.R., Costa-Neto S.F., Gentile R, & Melo R.C. (2016). Natural Schistosoma mansoni Infection in the Wild Reservoir Nectomys squamipes Leads to Excessive Lipid Droplet Accumulation in Hepatocytes in the Absence of Liver Functional Impairment. PLoS ONE, 11(11), e0166979.

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Quantitative Hepatic CD105 Immunohistochemistry

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For immunohistochemical staining of hepatic CD105, liver specimens were fixed in 10% formalin, paraffin-embedded and stained with goat anti-human endoglin/CD105 (R&D Systems, Inc. Canada) at 4°C overnight. Tissue was stained using anti-goat HRP-DAB and counterstained with hematoxylin. Staining was evaluated by Pannoramic Viewer (Histoquant; 3DHistech, Budapest, Hungary) and quantification (% of stained area) of IHC staining is expressed as mean±SEM.

Klein S., Hinüber C., Hittatiya K., Schierwagen R., Uschner F.E., Strassburg C.P., Fischer H.P., Spengler U, & Trebicka J. (2016). Novel Rat Model of Repetitive Portal Venous Embolization Mimicking Human Non-Cirrhotic Idiopathic Portal Hypertension. PLoS ONE, 11(9), e0162144.

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Hepatic alpha-SMA Immunohistochemistry

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Immunohistochemical stainings for hepatic αSMA were prepared using a cryostat. Cryo sections (4–6μm) of snap-frozen liver samples were fixed and incubated with mouse-anti-αSMA antibody (Sigma Aldrich, München, Germany). Thereafter, a biotinylated donkey-anti-rat secondary antibody was used (Abcam, Cambridge, UK). Sections were detected and quantified using computerized image capture device (Histoquant; 3DHistech, Budapest, Hungary). Results are expressed as mean±SEM.

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Histological Assessment of Liver Fibrosis

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For the detection of collagen fibers, liver specimen were fixed in 10% formalin, paraffin-embedded and stained in 0.1% Sirius-red in saturated picric acid (Chroma, Münster, Germany) using standard methods as previously described [11 (link)].
For immunohistochemical (IHC) staining of α-smooth muscle actin (αSMA), slides with sections were incubated with a mouse-anti-αSMA (clone 1A4; Sigma–Aldrich, St. Louis, USA) diluted 1:100 in Tris–buffered saline overnight. A secondary biotinylated rabbit-anti-mouse antibody, absorbed with rat serum (Dako, Glostrup, Denmark), was subsequently applied (1:300, 30 min) and complexed with streptavidin-conjugated alkaline phosphatase (1:500, 30 min; Dako). Finally, slides were developed with AEC (3-amino-9-ethylcarbazole) (15 min; Dako) and counterstained with hematoxylin. The amount of staining was evaluated by computational analysis (Histoquant; 3DHistech, Budapest, Hungary). Quantification (% of stained area) of IHC staining is expressed as mean±SEM.

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