Thermo autostainer 360

Manufactured by Thermo Fisher Scientific

The Thermo Autostainer 360 is a fully automated slide staining system designed for immunohistochemistry and in situ hybridization applications. It provides consistent and reproducible staining results through precise reagent and temperature control.

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Lab products found in correlation

Avidin d and biotin solutions, by Vector Laboratories (4 mentions) Biotinylated goat anti rabbit igg, by Vector Laboratories (3 mentions) Dako dab chromagen, by Agilent Technologies (3 mentions) Sigma citrate buffer, by Merck Group (3 mentions) Proteinase k, by Agilent Technologies (2 mentions) Vector horseradish peroxidase streptavidin, by Vector Laboratories (2 mentions) Biotinylated goat αrabbit igg, by Vector Laboratories (1 mentions) Goat anti rabbit igg, by Vector Laboratories (1 mentions) Dako dab chromogen, by Agilent Technologies (1 mentions)

Close spelling variants of this product

Autostainer 360 (13 citations) Autostainer (2 citations)

6 protocols using thermo autostainer 360

Immunohistochemical Detection of LASV

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Tissue sections were deparaffinized and rehydrated through xylene, graded ethanols, and then processed for IHC using the Thermo Autostainer 360 (ThermoFisher, Kalamazoo, MI). Slides were treated with Proteinase K for 5 min to unmask antigens (Dako, Carpenteria, CA #S3020) followed by sequential 15-min incubations with avidin D and biotin solutions (Vector, Burlingame, CA) to block endogenous biotin reactivity. Specific anti-LASV immunoreactivity was detected using an anti-LASV NP primary antibody (Zalgen Labs, Frederick MD #CLD4) at a 1:1,000 dilution for 60 min. Secondary antibody used was biotinylated goat anti-rabbit IgG (Vector Laboratories, Burlingame, CA #BA-1000) at 1:200 for 30 min followed by Horseradish Peroxidase Streptavidin, R.T.U (Vector) for 30 min. Slides were developed with Dako DAB chromagen (Dako, Carpenteria, CA #K3468) for 5 min and counterstained with hematoxylin for 30 s.

Cross R.W., Heinrich M.L., Fenton K.A., Borisevich V., Agans K.N., Prasad A.N., Woolsey C., Deer D.J., Dobias N.S., Rowland M.M., Lathigra R., Borrega R., Geisbert J.B., Garry R.F., Branco L.M, & Geisbert T.W. (2023). A human monoclonal antibody combination rescues nonhuman primates from advanced disease caused by the major lineages of Lassa virus. Proceedings of the National Academy of Sciences of the United States of America, 120(34), e2304876120.

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Immunohistochemical Detection of ZEBOV

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A partial necropsy was performed on all subjects. Tissue samples of major organs were collected for histopathologic and immunohistochemical examination, immersion-fixed in 10% neutral buffered formalin, and processed for histopathology as previously described^{27 (link),28 (link)}. For immunohistochemistry, specific anti-ZEBOV immunoreactivity was detected using an anti-ZEBOV VP40 primary antibody (IBT) at a 1:4000 dilution for 60 min. The tissues were processed for immunohistochemistry using the Thermo Autostainer 360 (ThermoFisher, Kalamazoo, MI). Secondary used was biotinylated goat anti-rabbit IgG (Vector Labs, Burlingame, CA #BA-1000) at 1:200 for 30 min followed by Vector Horseradish Peroxidase Streptavidin, R.T.U (Vector) for 30 min. Slides were developed with Dako DAB chromagen (Dako, Carpenteria, CA #K3468) for 5 min and counterstained with Harris Hematoxylin for 30 s.

Cross R.W., Bornholdt Z.A., Prasad A.N., Geisbert J.B., Borisevich V., Agans K.N., Deer D.J., Melody K., Fenton K.A., Feldmann H., Sprecher A., Zeitlin L, & Geisbert T.W. (2020). Prior vaccination with rVSV-ZEBOV does not interfere with but improves efficacy of postexposure antibody treatment. Nature Communications, 11, 3736.

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Immunohistochemical Detection of CCHFV

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Tissue sections were deparaffinized and rehydrated through xylene and graded ethanols. Slides went through heat antigen retrieval in a steamer at 95 °C for 20 mins in Sigma Citrate Buffer, pH6.0, 10× (Sigma Aldrich, St. Louis, MO). To block endogenous peroxidase activity, slides were treated with a 3% hydrogen peroxide and rinsed in distilled water. The tissue sections were processed for IHC using the Thermo Autostainer 360 (ThermoFisher, Kalamazoo, MI). Sequential 15 min incubations with avidin D and biotin solutions (Vector, Burlingame, CA) were performed to block endogenous biotin reactivity. Specific anti-CCHFV immunoreactivity was detected using a primary polyclonal rabbit-α-CCHFV-NP antibody (IBT BioServices, Rockville, MD) at a 1:3200 dilution for 60 mins. A secondary biotinylated goat-α-rabbit-IgG (Vector Laboratories, Burlingame, CA) at 1:200 dilution for 30 mins followed by Vector Horseradish Peroxidase Streptavidin, R.T.U (Vector) for 30 mins. Slides were developed with Dako DAB chromagen (Dako, Carpenteria, CA) for 5 mins and counterstained with Harris hematoxylin for 30 seconds. Tissue sections from uninfected mice were used as negative controls.

Rodriguez S.E., Cross R.W., Fenton K.A., Bente D.A., Mire C.E, & Geisbert T.W. (2019). Vesicular Stomatitis Virus-Based Vaccine Protects Mice against Crimean-Congo Hemorrhagic Fever. Scientific Reports, 9, 7755.

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Immunohistochemical Analysis of BDBV

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Necropsy was performed on all subjects in the BSL-4 facility. Tissue samples for histopathologic and immunohistochemical (IHC) examination were immersed in 10% neutral buffered formalin for at least 21 days, followed by a change of formalin, before removal from the BSL-4 laboratory. Inactivated tissue samples were processed in a BSL-1 laboratory. Tissue sections were deparaffinized and rehydrated through xylene and graded ethanols. Slides went through heat antigen retrieval in a steamer at 95°C for 20 minutes in Sigma citrate buffer, pH 6.0, 10 × (Sigma Aldrich). The tissue sections were processed for IHC using the Thermo Autostainer 360 (ThermoFisher). Specific anti-BDBV immunoreactivity was detected using an anti-BDBV GP primary antibody at a 1:2000 dilution for 60 minutes (IBT BioServices). Secondary antibody used was biotinylated goat anti-rabbit IgG (No. BA-1000; Vector Laboratories) at 1:200 for 30 minutes followed by Vector Streptavidin Alkaline Phosphatase at a dilution of 1:200 for 20 minutes (No. SA-5100; Vector Laboratories). Slides were developed with Bio-Red (No. BP-100-FR; Biopath Laboratories) for 7 minutes and counterstained with hematoxylin for 45 seconds.

Woolsey C., Strampe J., Fenton K.A., Agans K.N., Martinez J., Borisevich V., Dobias N.S., Deer D.J., Geisbert J.B., Cross R.W., Connor J.H, & Geisbert T.W. (2023). A Recombinant Vesicular Stomatitis Virus-Based Vaccine Provides Postexposure Protection Against Bundibugyo Ebolavirus Infection. The Journal of infectious diseases, 228(Suppl 7).

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Immunohistochemical Detection of Lassa Virus

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Tissue sections were deparaffinized and rehydrated through xylene, graded ethanols and then processed for IHC using the Thermo Autostainer 360 (ThermoFisher, Kalamazoo, MI). Slides were treated with Proteinase K for 5 min to unmask antigens (Dako, Carpenteria, CA #S3020) followed by sequential 15-min incubations with avidin D and biotin solutions (Vector, Burlingame, CA) to block endogenous biotin reactivity. Specific anti-LASV immunoreactivity was detected using an anti-LASV NP primary antibody (Zalgen Labs, Frederick MD #CLD4) at a 1:1000 dilution for 60 min. Secondary antibody used was biotinylated goat anti-rabbit IgG (Vector Laboratories, Burlingame, CA #BA-1000) at 1:200 for 30 min followed by Horseradish Peroxidase Streptavidin, R.T.U (Vector) for 30 min. Slides were developed with Dako DAB chromagen (Dako, Carpenteria, CA #K3468) for 5 min and counterstained with hematoxylin for 30 s.

Cross R.W., Fenton K.A., Woolsey C., Prasad A.N., Borisevich V., Agans K.N., Deer D.J., Dobias N.S., Fears A.C., Heinrich M.L., Geisbert J.B., Garry R.F., Branco L.M, & Geisbert T.W. (2024). Monoclonal antibody therapy protects nonhuman primates against mucosal exposure to Lassa virus. Cell Reports Medicine, 5(2), 101392.

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Immunohistochemical Staining for Nipah Virus

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Tissue sections were deparaffinized and rehydrated through xylene and graded ethanol washes. Slides went through heat antigen retrieval in a steamer at 95°C for 20 minutes in Sigma Citrate Buffer, pH 6.0, 10× (Sigma-Aldrich). To block endogenous peroxidase activity, slides were treated with 3% hydrogen peroxide and rinsed in distilled water. The tissue sections were processed for IHC using the Thermo Autostainer 360 (Thermo Fisher Scientific). Sequential 15-minute incubations with avidin D and biotin solutions (Vector Laboratories, SP-2001) were performed to block endogenous biotin reactivity. Specific anti-NiV immunoreactivity was detected using an anti-NiV m102.4 human monoclonal antibody (53 (link)) at a 1:4,000 dilution for 60 minutes. Secondary antibody was biotinylated goat anti-rabbit IgG (Vector Laboratories, BA-1000) at 1:200 for 30 minutes followed by Vector Horseradish Peroxidase Streptavidin (ready-to-use; Vector Laboratories, SA-5704) for 30 minutes. Slides were developed with Dako DAB chromogen (Dako, K3468) for 5 minutes and counterstained with hematoxylin for 45 seconds.

Woolsey C., Borisevich V., Fears A.C., Agans K.N., Deer D.J., Prasad A.N., O’Toole R., Foster S.L., Dobias N.S., Geisbert J.B., Fenton K.A., Cross R.W, & Geisbert T.W. (2023). Recombinant vesicular stomatitis virus–vectored vaccine induces long-lasting immunity against Nipah virus disease. The Journal of Clinical Investigation, 133(3), e164946.

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