A1rs confocal microscope system

Manufactured by Nikon

The Nikon A1RS confocal microscope system is a high-performance imaging platform designed for advanced microscopy applications. It features a resonant scanner for fast image acquisition, providing rapid scanning capabilities. The system is capable of capturing images with high resolution and sensitivity, making it suitable for a wide range of scientific and research applications.

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Lab products found in correlation

Ab53219, by Abcam (2 mentions) Af980, by R&D Systems (1 mentions) Normal rabbit igg, by Thermo Fisher Scientific (1 mentions) Fitc anti alpha smooth muscle actin, by Abcam (1 mentions) Alexa fluor 594 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Ab125884, by Abcam (1 mentions) Ab8211, by Abcam (1 mentions) Nb600 408, by Novus Biologicals (1 mentions) Af2519, by R&D Systems (1 mentions) Sc 271098, by Santa Cruz Biotechnology (1 mentions) In situ cell death detection kit, by Roche (1 mentions) Ab15580, by Abcam (1 mentions) Mca497, by Bio-Rad (1 mentions)

5 protocols using a1rs confocal microscope system

Multimodal Imaging of Aortic Thrombospondin-1

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Paraffin-embedded human AAA tissue or fresh frozen murine aortic sections were examined using Advanced Cell Diagnostics (ACD) RNAscope® Fluorescent Multiplex Reagent Kit (320850) with RNAscope® Probe against human THBS1 (320850), human CD68 (560591-C3), mouse Thbs1 (457891-C3), mouse Cd68 (316611-C2), mouse Pecam1 (316721). Probe against DapB (dihydrodipicolinate reductase, from Bacillus subtilis) was used as negative control (310043-C3), probe against Ubc (Mus musculus ubiquitin C) was used as positive control (310771-C3). Tissue pretreatment, probe hybridization and signal amplification were performed according to manufacturer’s instructions. For RNA-FISH combined with immunostaining, after signal amplification, tissue sections were blocked with 10% normal donkey serum. Alexa Fluor® 488 anti-mouse MHC II antibody (1:100, 107616, clone M5/114.15.2, BioLegend) were applied onto tissue sections, and incubated 2 hours at room temperature. Images were acquired with a Nikon A1RS confocal microscope system and analyzed using ImageJ software.

Yang H., Zhou T., Sorenson C.M., Sheibani N, & Liu B. (2020). Myeloid-derived Thrombospondin-1 contributes to abdominal aortic aneurysm through suppressing tissue inhibitor of metalloproteinases-1. Arteriosclerosis, thrombosis, and vascular biology, 40(12), e350-e366.

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Immunofluorescence Staining of TIMP1, MYH11 and F4/80

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PFA-fixed tissue sections or cells were blocked with 10% normal donkey serum and incubated at 4 °C overnight with primary antibodies anti- TIMP1 (1:100, AF980, R&D Systems), anti- MYH11 (1:200, ab53219, Abcam), PE anti-mouse F4/80 (1:100, 123110, clone BM8, BioLegend). Normal goat or rabbit IgG was used as negative controls. After several washes with PBS, sections were incubated with Alexa Fluor 488- or Alexa Fluor 594-conjugated secondary antibodies for 1 hour at room temperature. DAPI was used to stain nuclei. Images were acquired with a Nikon A1RS confocal microscope system and analyzed using ImageJ software.

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Immunofluorescence Analysis of Cell Signaling

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Tissue sections were fixed with 4% paraformaldehyde (PFA) for 10 min, followed by permeabilization with 0.1% Triton X-100 for 10 min at room temperature. Non-specific sites were blocked using 3% bovine serum albumin (BSA) for 1 h at room temperature. Tissue sections were incubated at 4 °C overnight in 0.3% BSA and 0.1% Tween 20 containing primary antibodies anti-phospho-MLKL Ser345 (1:1000, Cell Signaling Technology, Danvers, MA, USA, 37333) or anti-phospho-CaMKII Thr287 (1:100, generated and validated by Dr. Singer’s lab) [16 (link),17 (link)] and FITC Anti-alpha smooth muscle actin (1:500, Abcam, Waltham, MA, USA, ab8211). Normal rabbit IgG (Thermo Scientific, Waltham, MA, USA, 026102) was used for the negative controls. After washing three times in PBS, sections were incubated with Alexa Fluor 594-conjugated secondary antibodies (1:500, Thermo Scientific, Waltham, MA, USA) for 1 h at room temperature in the dark. DAPI was used to stain nuclei. Images were acquired with a Nikon A1RS confocal microscope system, and analyzed using ImageJ software.

Zhou T., DeRoo E., Yang H., Stranz A., Wang Q., Ginnan R., Singer H.A, & Liu B. (2021). MLKL and CaMKII Are Involved in RIPK3-Mediated Smooth Muscle Cell Necroptosis. Cells, 10(9), 2397.

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Multiparametric Immunofluorescence Analysis

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Tissue sections were fixed with 4% PFA, permeabilized with 0.1% Triton X-100, and blocked with 3% BSA. Sections were incubated at 4 °C overnight in 0.3% BSA containing primary antibodies: FITC anti-α-smooth muscle actin (ab8211, Abcam; 1:500), anti-Ly6G (127602, BioLegend; 1:800), anti-MYH11 (ab125884, Abcam, 1:200), anti-Fibronectin-1 (sc-271098, Santa Cruz, 1:100), anti-CD36 (AF2519, R&D systems, 1:100), anti-KLF2 (NBP2-45510, Novus Biologicals, 1:100), anti-ACTC1 (MAB93081-SP, R&D systems, 1:100), or anti-COL1A1 (NB600-408, Novus Biologicals, 1:100). After several washes with PBS, sections were incubated with Alexa Fluor 488- or Alexa Fluor 594-conjugated secondary antibodies for 1 h at room temperature. Negative control slides were stained with secondary antibody only. TUNEL staining was performed on fresh frozen sections using an In Situ Cell Death Detection Kit (Roche, Catalog #12 156 792 910) according to manufacture instructions. Co-staining with FITC anti-α-smooth muscle actin (ab8211, Abcam; 1:500) was performed. DAPI-containing mounting media (GBI Labs, Catalog #E19-100) was used as a counterstain. Images were acquired with a Nikon A1RS confocal microscope system.

DeRoo E., Zhou T., Yang H., Stranz A., Henke P, & Liu B. (2023). A vein wall cell atlas of murine venous thrombosis determined by single-cell RNA sequencing. Communications Biology, 6, 130.

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Multicolor Immunofluorescence Staining

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Tissue sections were fixed with 4% PFA (paraformaldehyde) for 10 minutes, followed by permeabilization with 0.1% Triton X-100 for 10 minutes at room temperature. Non-specific sites were blocked using 10% normal donkey serum for 1 hour at room temperature. Tissue sections were incubated at 4 °C overnight in antibody diluent (003218, ThermoFisher Scientific) containing primary antibodies anti-MYH11 (1:200, ab53219, Abcam), or anti- F4/80 (1:100, MCA497, Bio-Rad) and anti- Ki-67 (1:100, ab15580, Abcam). Normal rabbit and rat IgG isotype controls were used as negative controls. After washing three times in PBS, sections were incubated with Alexa Fluor 594- and Alexa Fluor 647-conjugated secondary antibodies (1:500, Invitrogen) for 1 hour at room temperature in the dark. DAPI was used to stain nuclei. Images were acquired with a Nikon A1RS confocal microscope system, and analyzed using ImageJ software. Positive areas were determined using image J; after auto-thresholding, areas with signal exceeding the threshold value were considered positive area.

Yang H., Zhou T., Stranz A., DeRoo E, & Liu B. (2021). Single-cell RNA sequencing reveals heterogeneity of vascular cells in early stage murine abdominal aortic aneurysm. Arteriosclerosis, thrombosis, and vascular biology, 41(3), 1158-1166.

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