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Innoscan 1100 al

Manufactured by Innopsys
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The InnoScan 1100 AL is a high-performance microarray scanner designed for DNA, protein, and small molecule array analysis. The instrument utilizes advanced optics and a powerful excitation laser to provide accurate and reproducible data. Key technical specifications include a maximum scanning resolution of 2 µm, a scan area of up to 24 x 76 mm, and compatibility with a wide range of microarray slide formats.

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Lab products found in correlation

Mapix, by Innopsys (5 mentions) Mapix 7, by Innopsys (3 mentions) Mapix 8, by Innopsys (3 mentions) Streptavidin alexa fluor 555, by Thermo Fisher Scientific (2 mentions) Lsm 710, by Zeiss (2 mentions) Protoarray v5, by Thermo Fisher Scientific (1 mentions) Pa055, by Thermo Fisher Scientific (1 mentions) Alexa fluor 647 goat anti human igg antibody, by Thermo Fisher Scientific (1 mentions) Dylight 550 goat anti gst, by Cayman Chemical (1 mentions) Genepix pro 6.0 analysis software, by Molecular Devices (1 mentions) Mapix software, by Innopsys (1 mentions) Prism 6, by GraphPad (1 mentions) Alexa fluor 488 nhs ester, by Thermo Fisher Scientific (1 mentions) Atto 532, by Merck Group (1 mentions) Slideh, by Schott (1 mentions) Goat anti mouse igg alexfluor 647, by Thermo Fisher Scientific (1 mentions) Contour gt, by Bruker (1 mentions) Genepix 4000b, by Molecular Devices (1 mentions) Fla 5100, by Fujifilm (1 mentions) Dylight 488, by Thermo Fisher Scientific (1 mentions)

21 protocols using innoscan 1100 al

1

Plasma Autoantibody Reactivity Profiling

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Plasma autoantibody reactivity was studied using full-length human protein arrays (ProtoArray v5.1, PAH05251020, Thermo Fisher) [40 (link), 41 ]. The patient and two healthy blood donors were investigated in the same experiment. Protein arrays were probed with plasma at a dilution of 1:2000, and otherwise following the manufacturer’s protocol. Protein arrays were first incubated with blocking buffer (PA055, Life Technologies) for 1 h, followed by 90-min incubation with plasma, and lastly, a 90-min incubation with detection antibodies: Alexa Fluor 647 goat anti-human IgG antibody (A21445, Thermo Fisher) at 1:2000 dilution and Dylight 550 goat anti-GST (#DY550011-13–001, Cayman Chemicals) at 1:10,000 dilution. The Innopsys InnoScan 1100 AL 3-channel ultra-high resolution microarray scanner was used for detection.
Abolhassani H., Landegren N., Bastard P., Materna M., Modaresi M., Du L., Aranda-Guillén M., Sardh F., Zuo F., Zhang P., Marcotte H., Marr N., Khan T., Ata M., Al-Ali F., Pescarmona R., Belot A., Béziat V., Zhang Q., Casanova J.L., Kämpe O., Zhang S.Y., Hammarström L, & Pan-Hammarström Q. (2022). Inherited IFNAR1 Deficiency in a Child with Both Critical COVID-19 Pneumonia and Multisystem Inflammatory Syndrome. Journal of Clinical Immunology, 42(3), 471-483.
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2

Biotinylated Viral Glycan Binding Assay

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Viruses were directly labeled with a biotin handle as previously described (Watanabe et al., 2014 (link)). Labeled viruses were diluted to 256 hemagglutinating units (HAU) in 1X PBS and applied directly to the slide surface for 1h. Following the initial incubation, arrays were washed, by dipping 3 times in 1X PBS and again 3 times in 1X PBS. Washed arrays were incubated with 2ug/mL streptavidin-AlexaFluor555 (LifeTechnologies) in 1X PBS, for 1h. Following detection, arrays were washed sequentially, by dipping, 3 times in 1X PBS, 3 times in 1X PBS and, finally, 3 times in deionized H2O. Washed arrays were dried by centrifugation and immediately scanned for AlexaFluor555 signal on an Innoscan 1100AL (Innopsys) confocal microarray scanner. Signal intensity from scanned arrays was collected using Mapix (Innopsys). Signal intensity was calculated for the mean signal intensity of 4 replicate spots for each printed glycan and graphed using Excel (Microsoft).
Herfst S., Zhang J., Richard M., McBride R., Lexmond P., Bestebroer T.M., Spronken M.I., de Meulder D., van den Brand J.M., Rosu M.E., Martin S.R., Gamblin S.J., Xiong X., Peng W., Bodewes R., van der Vries E., Osterhaus A.D., Paulson J.C., Skehel J.J, & Fouchier R.A. (2020). Hemagglutinin traits determine transmission of avian A/H10N7 influenza virus between mammals. Cell host & microbe, 28(4), 602-613.e7.
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3

Microarray Data Normalization Protocol

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Slides were scanned at 5 μm resolution with an InnoScan 1100 AL fluorescence scanner (Innopsys, Carbonne, France). Image analysis was carried out with Genepix Pro 6.0 analysis software (Molecular Devices Corporation, Union City, CA) as previously described [14 (link),15 (link)]. Spots were defined as circular features with a diameter of 90 μm. Local background subtraction (median background) was performed and the background-subtracted median pixel intensity feature was used. To minimize the impact of noise on our comparisons, spots with intensity lower than 150 (1/2 the typical background signal when analyzing IgM and IgG at 1:50) were considered too low to be measured accurately and were set to 150. The average of duplicate spots was calculated to obtain a normalized value to the reference samples. A log-transformed (base 2) was applied for each slide, and the final data value was obtained from the normalized average of data from both wells for a given macaque sample. Full microarray data can be found in the Supplemental Table S1.
Nanno Y., Sterner E., Gildersleeve J.C., Hering B.J, & Burlak C. (2019). Profiling Natural Serum Antibodies of Nonhuman Primates with a Carbohydrate Antigen Microarray. Xenotransplantation, 27(2), e12567.
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4

Peptide Microarray Fabrication and Antibody Binding Analysis

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Peptide microarrays were designed using ArrayNinja41 (link) and fabricated using an Aushon 2470 microarrayer as described previously26 (link). Antibodies were diluted in Array Hybridization Buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.6, 5% BSA, 0.1% Tween-20) and incubated over each microarray slide for 1 h at 4 °C. Slides were washed in PBS, incubated with fluorescently labeled secondary antibodies (Life Technologies A-21244 or A-21235), washed in PBS, and scanned using an Innopsys InnoScan 1100AL microarray slide scanner. Microarray images were analyzed using ArrayNinja software.
Berryhill C.A., Hanquier J.N., Doud E.H., Cordeiro-Spinetti E., Dickson B.M., Rothbart S.B., Mosley A.L, & Cornett E.M. (2023). Global lysine methylome profiling using systematically characterized affinity reagents. Scientific Reports, 13, 377.
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5

Glycan Array Analysis of Antibody Binding to Influenza NA Antigen

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The Sh2 N9 NA tetramer with ectodomain plus stalk and His6-tag was diluted to 20 pg/mL and incubated with antibody IgG at a molar ratio of one NA protomer to four antibody IgGs for 1 hour at room temperature (~ 22 °C) in 100 mM imidazole-malate pH 6.15, 150 mM NaCl, 10 mM CaCl2, and 0.02% NaN3. The mixtures were then applied to glycan arrays (see Data S1 for glycan list) (Peng et al., 2017 (link)) for 1 hour. Then each array was washed to remove the NA solution by dipping 3 times with 1× PBS + 0.05% Tween, pH 7.4 at room temperature. Following washing, 250 μL of a pre-complexed solution of biotinylated Erythrina cristagalli lectin (ECA) (10 μg/mL, VectorLabs) + Streptavidin-AlexaFluor555 (2 μg/mL, Thermo Fisher Scientific) was applied directly to the array surface and incubated for 2 hours. Following incubation, ECA-Streptavidin solution was removed by dipping 3 times with 1× PBS + 0.05% Tween and, subsequently, by dipping 3 times in 1× PBS and then 3 times in distilled H2O. Washed slides were dried by centrifugation and scanned on an Innoscan1100AL (Innopsys) confocal slide scanner for 532 emission. Image data were stored as a TIFF image and signal data were collected using Mapix (Innopsys) imaging software. Collected signal data were processed to determine the averaged (mean signal minus mean background) values of 4 replicate spots on the array for each unique printed glycan.
Zhu X., Turner H.L., Lang S., McBride R., Bangaru S., Gilchuk I.M., Yu W., Paulson J.C., Crowe JE J.r., Ward A.B, & Wilson I.A. (2019). Structural basis of protection against H7N9 influenza virus by human anti-N9 neuraminidase antibodies. Cell host & microbe, 26(6), 729-738.e4.
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6

Fluorescence Image Analysis Protocol

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Fluorescence scanning was performed on the Innoscan 1100 AL (Innopsys, Carbonne, France) at 647 nm, gain 3, laser power low, and a resolution of 1 μm/pixel. The resulting 16-bit grayscale images were analyzed using Mapix (Innopsys, Carbonne, France). The median gray scale value of all pixels in a spot was subtracted by the median gray scale value of at least 5 background control areas (spot size of 80–90 µm each), located in the vicinity of the spot of interest.
Palermo A., Thelen R., Weber L.K., Foertsch T., Rentschler S., Hackert V., Syurik J, & Nesterov-Mueller A. (2019). Vertical Scanning Interferometry for Label-Free Detection of Peptide-Antibody Interactions. High-Throughput, 8(2), 7.
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7

Fluorescence Microscopy Protocols

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Images of patterns were acquired using InnoScan 1100 AL fluorescence scanner (Innopsys) through simultaneous acquisition at 3 different wavelengths (488 nm, 532 nm and 635 nm). The fluorescence images were analysed for quantification of fluorescence intensity using Mapix software (Innopsys) or ImageJ software. Coefficients of variation (CV) were calculated as the quotient of standard deviation over the mean of fluorescence intensities. Images of cells were acquired by confocal microscopy (Zeiss, LSM 710, 63 x objective).
Foncy J., Estève A., Degache A., Colin C., Dollat X., Cau J.C., Vieu C., Trévisiol E, & Malaquin L. (2018). Dynamic inking of large-scale stamps for multiplexed microcontact printing and fabrication of cell microarrays. PLoS ONE, 13(8), e0202531.
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8

Quantifying Glycan-Antibody Interactions

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Glycan-antibody binding on the microarray was quantified using an InnoScan 1100 AL fluorescence scanner (Innopsys; Chicago, IL). Images were analyzed using GenePix Pro 7.0. Fluorescent intensity values calculated in GenePix Pro 7.0 were plotted using GraphPad Prism 6.0. Each neoglycoprotein was printed in duplicate in each well, and each antibody was assayed in a minimum of 2 independent array experiments for each concentration tested. The final intensity values for each antibody-neoglycoprotein interaction (Figures 3, S3 and S4) were calculated from the average of corresponding spots in each of the two (or more) wells (duplicate spots in each of two or more wells; total of 4 or more spots). Apparent KD values were calculated using a non-linear regression, one site binding (hyperbola) fitting model following the method of MacBeath (Gordus and MacBeath, 2006 (link)). The list of array components and full microarray data will be publicly available in the National Center for Biotechnology Information’s (NCBI) Gene Expression Omnibus (GEO) (Edgar et al., 2002 ) via GEO Series accession number (GSE100438).
Sterner E., Peach M.L., Nicklaus M.C, & Gildersleeve J.C. (2017). Therapeutic Antibodies to Ganglioside GD2 Evolved from Highly Selective Germline Antibodies. Cell reports, 20(7), 1681-1691.
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9

Glycan Microarray Analysis of Lectin Specificity

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The fine carbohydrate specificity of the lectin toward mammalian glycans and bacterial polysaccharides was determined using a glycan microarray constructed by Semiotik LLC (Moscow, Russia). Pure GYL was labeled with the biotin according to the product manual. The array was composed of more than 600 tested glycans, each in 6 replicates. The amount of biotinylated GYL bound to each glycan was determined after adding FITC-labeled streptavidin. Slides were scanned using an InnoScan1100 AL (Innopsys, Carbonne, France) equipped with a 488 nm laser at 100 PMT and a high-power laser mode. The data were processed by fixed 100 μm diameter ring method using Mapix 7.3.1 and Mapix 8.2.2 software (Innopsys, France). Data were presented as the relative fluorescence units (RFU) median of six spot replicates. The signals with fluorescence intensity over the background value by a factor of five passed as significative.
Mizgina T.O., Baldaev S.N., Likhatskaya G.N., Molchanova V.I., Kokoulin M.S., Filshtein A.P., Rogozhin E.A., Chikalovets I.V., Isaeva M.P, & Chernikov O.V. (2023). Molecular Cloning and Characteristics of a Lectin from the Bivalve Glycymeris yessoensis. Marine Drugs, 21(2), 55.
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10

Glycan microarray profiling of lectin

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The fine carbohydrate specificity of the lectin towards mammalian glycans and bacterial polysaccharides was determined using a glycan microarray constructed by Semiotik LLC (Moscow, Russia). Pure GYL-R was labeled with biotin according to the product manual. The array was composed of more than 600 tested glycans, each in 6 replicates. The amount of biotinylated GYL-R bound to each glycan was determined after adding FITC-labeled streptavidin. Slides were scanned with an InnoScan1100 AL (Innopsys, Carbonne, France) with a 488 nm laser at 100% PMT (photomultiplier) gain and high laser power mode. The obtained data were processed using the software Mapix 7.3.1 and Mapix 8.2.2 (Innopsys, France) and the fixed 100 μm-diameter ring method. Data are reported as the relative fluorescence units (RFU) median of six spot replicates. Signals with fluorescence intensity exceeding the background value by a factor of five were considered significant.
Mizgina T.O., Chikalovets I.V., Bulanova T.A., Molchanova V.I., Filshtein A.P., Ziganshin R.H., Rogozhin E.A., Shilova N.V, & Chernikov O.V. (2023). New l-Rhamnose-Binding Lectin from the Bivalve Glycymeris yessoensis: Purification, Partial Structural Characterization and Antibacterial Activity. Marine Drugs, 22(1), 27.
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