Airyscan

Manufactured by Zeiss

Sourced in Germany, United States, United Kingdom, Japan

The Airyscan is a detector system designed for confocal microscopy. It employs an array of small detectors to capture a high-resolution image, providing improved signal-to-noise ratio and faster image acquisition compared to traditional point detectors.

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Lab products found in correlation

Lsm 880, by Zeiss (324 mentions) Lsm 800, by Zeiss (57 mentions) Lsm 880 confocal microscope, by Zeiss (45 mentions) Lsm 780, by Zeiss (18 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (17 mentions) Zen software, by Zeiss (14 mentions) Lsm 710, by Zeiss (14 mentions) Lsm800 confocal microscope, by Zeiss (14 mentions) 4 6 diamidino 2 phenylindole (dapi), by Beyotime (11 mentions) Hoechst 33342, by Thermo Fisher Scientific (11 mentions) Lsm 880 microscope, by Zeiss (8 mentions) Triton x 100, by Merck Group (8 mentions) Zen blue software, by Zeiss (8 mentions) Zen 2.3 sp1 fp3, by Zeiss (7 mentions) Zen imaging software, by Zeiss (7 mentions) Zen 2, by Zeiss (7 mentions) Vectashield antifade mounting medium, by Vector Laboratories (6 mentions) Lsm 900, by Zeiss (5 mentions) Fluorescent in situ hybridization kit, by RiboBio (5 mentions) Live dead cell imaging kit, by Thermo Fisher Scientific (5 mentions)

486 protocols using airyscan

High-Resolution Imaging of Nociceptive Fibers

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Paw skin, DRGs, cornea, and spinal cord sections were imaged using an LSM 880 confocal microscope with AiryScan (Zeiss). The paw skin and cornea were imaged with the ×63 oil immersion objective using only the 568 nm channel. Z-stacking was conducted, with a step interval of 0.5 μm. An area of 8 tiles (135 × 260 μm) was imaged for all sections. DRGs were imaged with the 10× objective using 4 channels. Z-stacking was conducted with a step interval of 1 μm. An area of 12 tiles (1422 × 1896 μm) was imaged for all sections. The spinal cord was imaged with a ×20 objective using the 568 nm channel. Z-stacking was conducted with a step interval of 1 μm. An area of 15 tiles (711 × 1185.5 μm) was imaged for all sections. A total of 8 to 10 sections were imaged per animal.
For the high-resolution analysis of nociceptive fiber architecture in the glabrous paw skin and cornea, imaging using the AiryScan mode was conducted using Zeiss ×63/1.40 oil DIC f/ELYRA objective and the AiryScan super-resolution (SR) module with 32-channel hexagonal array GaAsP detector for LSM (Zeiss) using 651 nm lasers. Stacks of 20 to 30 optical sections (170 nm step) were acquired, and AiryScan SR image stacks were reconstructed using ZEN Black software (Zeiss). Images were analyzed using ZEN Blue software (Zeiss) and ImageJ (NIH).

Wong C., Barkai O., Wang F., Perez C.T., Lev S., Cai W., Tansley S., Yousefpour N., Hooshmandi M., Lister K.C., Latif M., Cuello A.C., Prager-Khoutorsky M., Mogil J.S., Séguéla P., De Koninck Y., Ribeiro-da-Silva A., Binshtok A.M, & Khoutorsky A. (2022). mTORC2 mediates structural plasticity in distal nociceptive endings that contributes to pain hypersensitivity following inflammation. The Journal of Clinical Investigation, 132(15), e152635.

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High-Resolution 3D Imaging of Drosophila Embryos

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Fixed Drosophila embryos mounted in ProLong Gold mounting media (Molecular Probes, Eugene, OR) were imaged on a Zeiss LSM 880 confocal microscope with Airyscan (Carl Zeiss Microscopy, Jena, Germany) using 3D Airyscan in SR mode to obtain images with 1.7-fold higher resolution compared to diffraction-limited confocal imaging (Sheppard et al., 2013 (link)) (method supplements: imaging setup for Airyscan). Images presented in the figures were processed with ImageJ (Schindelin et al., 2015 (link)).

Tsai A., Muthusamy A.K., Alves M.R., Lavis L.D., Singer R.H., Stern D.L, & Crocker J. (2017). Nuclear microenvironments modulate transcription from low-affinity enhancers. eLife, 6, e28975.

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Live-Cell Imaging of SNARE Complexes

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Live-cell imaging was performed using a Zeiss LSM 980 with Airyscan (Carl Zeiss). 48 h after transfection of a plasmid combination of nYFP-tagged VAMP4 with nYFP-tagged STX7, cYFP-tagged STX7, nYFP-tagged STX8, cYFP-tagged STX8, nYFP-tagged VTI1B, or cYFP-tagged VTI1B, cells were imaged in RPMI 1640 medium at 37°C in an atmosphere with 5% CO₂ using an 100× (NA = 1.40) objective and immersion oil optimized for 30°C (Carl Zeiss). Airyscan processing was performed using the Airyscan module in ZEN software (Carl Zeiss).

Li M., Feng F., Feng H., Hu P., Xue Y., Xu T, & Song E. (2022). VAMP4 regulates insulin levels by targeting secretory granules to lysosomes. The Journal of Cell Biology, 221(10), e202110164.

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Live Imaging of Neuron ER Morphology

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Neurons were imaged live on a Zeiss LSM 880 with AiryScan in the Weill Cornell Microscopy and Image Analysis Core Facility. Samples were imaged with a 40X, 1.3 NA oil objective. AiryScan deconvolution was performed in the Zeiss Zen software to improve resolution and reveal the ER morphology.

Farrell R.J., Bredvik K.G., Hoppa M.B., Hennigan S.T., Brown T.A, & Ryan T.A. (2024). A ratiometric ER calcium sensor for quantitative comparisons across cell types and subcellular regions. bioRxiv.

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Live-cell imaging of HaloTag7-labeled cells

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Live-cell imaging was performed using LSM880 with Airyscan (Zeiss) equipped with a 100 × 1.46 alpha-Plan-Apochromat oil immersion lens and Immersol^TM 518 F/37 °C (444970-9010-000, Zeiss). The day before imaging, cells were seeded on a glass bottom dish (627870, Greiner bio-one) with growth medium without phenol red. During live-cell imaging, the dish was mounted in a chamber (STXG-WSKMX-SET, TOKAI HIT) to maintain the incubation conditions at 37 °C and 5% CO₂. Images were acquired at intervals of 6 s, analyzed and Airyscan processed with Zeiss ZEN 2.3 SP1 FP3 (black, 64 bit) (ver. 14.0.21.201) and Fiji (ver. 2.0.0-rc-69/1.52p). For HaloTag7 staining before live-cell imaging, cells were cultured in medium containing HaloTag7 ligand at 37 °C for 30 min in a 5% CO₂ incubator.

Kemmoku H., Takahashi K., Mukai K., Mori T., Hirosawa K.M., Kiku F., Uchida Y., Kuchitsu Y., Nishioka Y., Sawa M., Kishimoto T., Tanaka K., Yokota Y., Arai H., Suzuki K.G, & Taguchi T. (2024). Single-molecule localization microscopy reveals STING clustering at the trans-Golgi network through palmitoylation-dependent accumulation of cholesterol. Nature Communications, 15, 220.

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Confocal Imaging of Subcellular Localization

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Confocal imaging was conducted to evaluate whether mGold fused to different organelle/cytoskeleton-localization tags/proteins would produce fluorescence with the expected pattern of subcellular localization. HeLa cells were transfected with FuGENE as described above (see the “preparation of human cells” section). Cells were placed on the glass-bottom 24-well plates without poly-l-lysine coating because we observed that poly-l-lysine unexpectedly seems to promote detachment with HeLa cells, producing rounder cells after the PBS wash step. Transfected HeLa cells were washed with PBS and resuspended in HBSS supplemented with 10 mM HEPES. Laser-scanning confocal images were obtained using a high-speed confocal microscope (LSM880 with Airyscan, Zeiss) driven by the Zen software (version 2.3 SP1, Zeiss). Images were acquired with a 40× 1.1-NA water immersion objective (LD C-Apochromat Korr M27, Zeiss), a 488 nm argon laser (LGK7812, Lasos) at 3% power, and a per-pixel dwell time of 2 μs. Emission light was filtered using a multipass beamsplitter (MBS 488/561/633, Zeiss) and acquired with a 32 channel GaAsP detector (Airyscan, Zeiss) with a detector gain of 740 and 1–Airy unit pinhole size. To increase the signal-to-noise ratio, two scans were performed and averaged for each image.

Lee J., Liu Z., Suzuki P.H., Ahrens J.F., Lai S., Lu X., Guan S, & St-Pierre F. (2020). Versatile phenotype-activated cell sorting. Science Advances, 6(43), eabb7438.

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Quantifying Microglial CD68 Levels

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Z-stacks were collected on an LSM 880 confocal microscope with AiryScan (Zeiss) on Superresolution mode and a 63x objective (NA 1.4). Laser power and gain were consistent across each image. AiryScan processing was performed in Zen software (Zeiss) at a setting of 6 (“optimal” setting). Images were analyzed using Imaris software (Bitplane) by creating a 3D surface rendering of individual microglia, thresholded to ensure microglia processes were accurately reconstructed, and maintained consistent thereafter. Microglia rendering was used to mask and render the CD68 channel within each microglia. CD68 volume per microglia was then calculated as the total volume of masked CD68 volume within the masked GFP volume.

Escoubas C.C., Dorman L.C., Nguyen P.T., Lagares-Linares C., Nakajo H., Anderson S.R., Cuevas B., Vainchtein I.D., Silva N.J., Xiao Y., Lidsky P.V., Wang E.Y., Taloma S.E., Nakao-Inoue H., Schwer B., Andino R., Nowakowski T.J, & Molofsky A.V. (2023). Type I interferon responsive microglia shape cortical development and behavior. bioRxiv.

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Live-cell Imaging of GPCR Trafficking

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Transfection was performed using a mixture of 500 ng mVenus–β-arrestin 2 and 200 ng Flag–PTH1R plasmids (per well in a 6-cm dish). After incubation for 1 day, the transfected cells were collected and reseeded on a 35-mm, collagen-coated glass bottom dish (Matsunami). After 1 day, medium was changed to DMEM without phenol-red and FBS (starvation buffer). After 1 h of incubation, the cells were incubated with Alexa-647-labelled (1:2,000) Flag-M1 antibody for 1 h, washed once in the starvation buffer and set on the confocal microscope. Live-cell imaging was performed using LSM880 with Airyscan (Zeiss) equipped with a ×100/1.46 alpha-Plan-Apochromat oil-immersion lens and ImmersolTM 518F/37 °C (444970-9010-000, Zeiss). During live-cell imaging, the dish was mounted in a chamber (STXG-WSKMX-SET, TOKAI HIT) to maintain the incubation conditions at 37 °C and 5% CO₂. We took dual-colour time-lapse images with the following settings: time interval of 5 min; total time of 35 min. The 200 µl ligand solution in 0.01% BSA-HBSS was added between time points 1 and 2. Acquired serial images were Airyscan processed using Zeiss ZEN 2.3 SP1 FP3 (black, 64 bit) (v.14.0.21.201). Co-localization analysis was performed using Fiji (v.2.0.0-rc-69/1.52p).

Kobayashi K., Kawakami K., Kusakizako T., Tomita A., Nishimura M., Sawada K., Okamoto H.H., Hiratsuka S., Nakamura G., Kuwabara R., Noda H., Muramatsu H., Shimizu M., Taguchi T., Inoue A., Murata T, & Nureki O. (2023). Class B1 GPCR activation by an intracellular agonist. Nature, 618(7967), 1085-1093.

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Immunofluorescence Microscopy of Extracellular and Intracellular Parasites

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Extracellular parasites were collected and purified as previously (Liu et al., 2014 (link)). Parasites were washed once with buffer A with glucose (BAG, 116 mM NaCl, 5.4 mM KCl, 0.8 mM MgSO₄, 5.5 mM glucose, and 50 mM HEPES, pH 7.4), and an aliquot of 2 × 10⁴ parasites was overlaid on a coverslip previously treated with poly-L-Lysine. Intracellular tachyzoites were grown on hTERT cells on coverslips. Both extracellular and intracellular parasites were fixed with 3% paraformaldehyde for 20 min at room temperature (RT), permeabilized with 0.3% Triton X-100, blocked with 3% bovine serum albumin (BSA), and exposed to primary antibodies (Ratα-HA 1:100). The secondary antibodies used were goat-αrat Alexa Fluor 488 (Life Technologies) at a 1:1000 dilution. For co-localization studies, we used α-Sag1 (1:1000) as membrane marker and α-TgSERCA as ER marker (1:1000). Slides were examined using an Olympus IX-71 inverted fluorescence microscope with a photometric CoolSNAP HQ charge-coupled device (CCD) camera driven by DeltaVision software (Applied Precision, Seattle, WA). Super-resolution images were imaged with a 63× oil (NA 1.4) objective on an 880-laser scanning microscope with Airyscan (Zeiss, Germany) with a 2× zoom. Airyscan images were process with the Zen Black Software (Zeiss, Germany).

Márquez-Nogueras K.M., Hortua Triana M.A., Chasen N.M., Kuo I.Y, & Moreno S.N. (2021). Calcium signaling through a transient receptor channel is important for Toxoplasma gondii growth. eLife, 10, e63417.

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Live-cell Imaging of HaloTag-Conjugated Proteins

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The day before imaging, cells were seeded on a glass-bottom dish (627870, Greiner Bio-One). The medium was changed to DMEM^gfp-2 (MC102, Evrogen) containing 10% FBS, PSG and rutin (20 µg ml⁻¹) (30319-04, Nacalai Tesque) before imaging. HaloTag SaraFluor 650T ligand was added to the medium for 10 min before live-cell imaging to visualize HaloTag-conjugated protein. Live-cell imaging was performed using LSM880 with Airyscan (Zeiss) equipped with a 100 × 1.46 alpha-Plan-Apochromat oil immersion lens and Immersol 518 F/37 °C (444970-9010-000, Zeiss). During live-cell imaging, the dish was mounted in a chamber (STXG-WSKMX-SET, TOKAI HIT) to maintain the incubation conditions at 37 °C and 5% CO₂. Acuired images were Airyscan processed with Zeiss ZEN 2.3 SP1 FP3 (black, 64 bit) (version 14.0.21.201) and analysed with Fiji (version 2.1.0/1.53c).

Kuchitsu Y., Mukai K., Uematsu R., Takaada Y., Shinojima A., Shindo R., Shoji T., Hamano S., Ogawa E., Sato R., Miyake K., Kato A., Kawaguchi Y., Nishitani-Isa M., Izawa K., Nishikomori R., Yasumi T., Suzuki T., Dohmae N., Uemura T., Barber G.N., Arai H., Waguri S, & Taguchi T. (2023). STING signalling is terminated through ESCRT-dependent microautophagy of vesicles originating from recycling endosomes. Nature Cell Biology, 25(3), 453-466.

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