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A1 confocal microscope system

Manufactured by Nikon
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Sourced in Japan, United States, Germany

The Nikon A1 is a confocal microscope system designed for high-resolution imaging of biological samples. It features a laser scanning unit, a motorized stage, and advanced imaging software to capture detailed, three-dimensional images of cells, tissues, and other microscopic specimens.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Merck Group (4 mentions) Alexa488 wheat germ agglutinin, by Thermo Fisher Scientific (3 mentions) Smz1000 stereomicroscope, by Nikon (3 mentions) Rabbit anti smyd3, by Abcam (2 mentions) Anti mouse cd4, by BD (2 mentions) Alexa fluor 568 goat anti rabbit, by Thermo Fisher Scientific (2 mentions) Anti mouse alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Alexa fluor 488 conjugated secondary antibody, by Thermo Fisher Scientific (2 mentions) Nis elements 4, by Nikon (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Rhodamine phalloidin, by Thermo Fisher Scientific (2 mentions) Dapi dilactate, by Thermo Fisher Scientific (2 mentions) Eclipse 90i, by Nikon (2 mentions) Fluorescent in situ hybridization kit, by GenePharma (1 mentions) Ab5392, by Abcam (1 mentions) Ab4674, by Abcam (1 mentions) Ab5922, by Merck Group (1 mentions) Hoechst 33342, by Dojindo Laboratories (1 mentions) Mms 435p, by Fortrea (1 mentions) Nis element viewer software, by Nikon (1 mentions)

47 protocols using a1 confocal microscope system

1

Immunofluorescence Staining of iTreg Cells

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Cell suspension from naïve and iTreg skewing cells stained as described 45 (link) using mouse anti-CD4 (BD-Pharmigen), rabbit anti-Smyd3 (Abcam), goat anti-rabbit Alexa Fluor 568, and anti-mouse Alexa Fluor 488 (Invitrogen). The stained tissue sections were analyzed using a Nikon A1 confocal microscope system (Nikon Instruments).
de Almeida Nagata D.E., Ting H.A., Cavassani K.A., Schaller M.A., Mukherjee S., Ptaschinski C., Kunkel S.L, & Lukacs N.W. (2015). Epigenetic control of Foxp3 by SMYD3 H3K4 histone methyltransferase controls iTreg development and regulates pathogenic T cell responses during pulmonary viral infection. Mucosal immunology, 8(5), 1131-1143.
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2

Fluorescent In Situ Detection of circAFF1 and miRNA-516b

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A Cy3-labeled circAFF1 probe and a Cy5-labeled miRNA-516b probe were designed and synthesized by GenePharma (Shanghai, China). After the cells were fixed with 4% paraformaldehyde and treated with Triton-X100, the fluorescently labeled probes were incubated with the cells. After washing with 2 × SSC buffer, DAPI was added to stain the nuclear DNA. The signals of the probe were detected by the Fluorescent In Situ Hybridization Kit (GenePharma, China) in accordance with the manufacturer’s protocols. All images were acquired on Nikon A1 Confocal Microscope system (Nikon, Japan).
Wang H.G., Yan H., Wang C., Li M.M., Lv X.Z., Wu H.D., Fang Z.H., Mo D.L., Zhang Z.Y., Liang B., Lai K.G., Bao J.Y., Yang X.J., Zhao H.J., Chen S., Fan Y.M, & Tong X.G. (2020). circAFF1 Aggravates Vascular Endothelial Cell Dysfunction Mediated by miR-516b/SAV1/YAP1 Axis. Frontiers in Physiology, 11, 899.
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3

Immunofluorescence Staining of hiPSC-Neurons

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After fixation with 4% paraformaldehyde (PFA) at room temperature for 30 min, hiPSC-neurons were permeabilized with 0.1% (vol/vol) Triton X-100 for 5 min, followed by treatment with blocking solution (5% [vol/vol] goat serum and 1% [wt/vol] BSA in PBS) for 60 min. After blocking, the cells were incubated with primary antibody (mouse monoclonal anti-β3 tubulin antibody [1:500] [MMS-435P, Covance, Princeton, NJ, USA], chicken polyclonal anti-GFAP antibody [1:400] [ab4674, Abcam, Cambridge, UK], rabbit polyclonal anti-Nestin antibody [1:200, AB5922, Millipore, Land Hessen, Germany], or chicken polyclonal anti-MAP2 antibody [1:5000, AB5392, Abcam, Cambridge, UK]) in blocking solution at 4°C for 24 h. After washing, the cells were incubated with solution containing Alexa Fluor 488-conjugated secondary antibody (1:500) (Invitrogen, Waltham, MA, USA) and Hoechst 33342 (1:200) (34607951, Dojindo, Kumamoto, Japan) at room temperature for 3 h. Cultured rat hippocampal neurons were counted in fluorescence images obtained and analyzed on a Nikon A1 confocal microscope system (Nikon, Tokyo, Japan).
Sato K., Takahashi K., Shigemoto-Mogami Y., Chujo K, & Sekino Y. (2016). Glypican 6 Enhances N-Methyl-D-Aspartate Receptor Function in Human-Induced Pluripotent Stem Cell-Derived Neurons. Frontiers in Cellular Neuroscience, 10, 259.
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4

Autophagy Monitoring in Prostate Cancer Cells

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PC-3, LNCaP and C4–2 cells stably transfected with mCherry-Wassabi-LC3B were seeded into glass bottom cell culture dishes with 1 × 105 cells per dish. After treatment, the cells were washed with PBS three times and examined under a Nikon A1 confocal microscope system (Nikon, Japan). Images were processed with NIS Element Viewer software (Nikon). Twenty cells were randomly selected to evaluate the average number of mCherry-Wassabi-LC3B puncta per cell.
Zhao F., Huang W., Zhang Z., Mao L., Han Y., Yan J, & Lei M. (2015). Triptolide induces protective autophagy through activation of the CaMKKβ-AMPK signaling pathway in prostate cancer cells. Oncotarget, 7(5), 5366-5382.
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5

Golgi-Cox Staining and Spine Analysis

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Mouse forebrain was harvested and stained with Golgi-Cox solution (FD Rapid GolgiStain Kit PK401, FD NeuroTechnologies, Inc. Columbia, MD) following the manufacturer’s directions. Coronal sections (150 μm) were cut using a vibratome (Precisionary Instruments LLC, Greenville, NC), slide mounted, and images were obtained at 100X (Nikon A1 Confocal Microscope System, Nikon Instruments, Inc., Melville, NY). The primary somatosensory cortex (barrel cortex) was defined in accordance with atlases of the developing and adult mouse brain and 6-7 sequential sections were imaged for each animal (33 , 34 ). Stack images were acquired (NIS Elements Confocal Microscope Imaging Software, Nikon Instruments, Inc., Melville, NY) and consisted of 150 optical sections, separated axially by 1 μm with a resolution of 0.1 μm/pixel. Images were analyzed (ImageJ, NIH) in a blinded manner and spines on apical dendrites of layer V, localized 50 to 100 μm from the soma of neurons, were counted and morphologically characterized as previously described (35 (link)).
Griffiths K.K., Wang A., Wang L., Tracey M., Kleiner G., Quinzii C.M., Sun L., Yang G., Perez-Zoghbi J.F., Licznerski P., Yang M., Jonas E.A, & Levy R.J. (2020). Inefficient Thermogenic Mitochondrial Respiration Due to Futile Proton Leak in a Mouse Model of Fragile X Syndrome. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34(6), 7404-7426.
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6

Ebola VLP Internalization in HepG2 Cells

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HepG2 cells were transfected with TPC2-EGFP and LAMP1-BFP plasmids using Lipofectamine 3000 (Invitrogen, US). At 24 hpt, HepG2 cells were pretreated with DMSO, 10 μM tamoxifen, clomiphene, or U18666a for 1 h. HepG2 cells were then incubated with Ebola VLP for 2 h, and then fixed. The cells were mounted and then imaged by the Nikon A1 Confocal Microscope System (Nikon, Japan).
Fan H., Du X., Zhang J., Zheng H., Lu X., Wu Q., Li H., Wang H., Shi Y., Gao G., Zhou Z., Tan D.X, & Li X. (2017). Selective inhibition of Ebola entry with selective estrogen receptor modulators by disrupting the endolysosomal calcium. Scientific Reports, 7, 41226.
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7

Immunofluorescence Analysis of MUC1 in Pancreatic Cancer

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Pancreatic cancer cells were seeded on poly-L-lysine-coated glass coverslips in 24-well culture plates. After 72 h, cells were blocked with 3% BSA, and incubated with HzMUC1 antibody for 1 h on ice, washed with PBS containing 1% BSA, and incubated with Alexa Flour 488-conjugated goat anti-human IgG antibody (Jackson ImmunoResearch) for 30 min. All samples were mounted and observed under the Nikon A1 confocal microscope system (Nikon, Tokyo, Japan).
Wu G., Li L., Liu M., Chen C., Wang G., Jiang Z., Qin Y., He L., Li H., Cao J, & Gu H. (2022). Therapeutic effect of a MUC1-specific monoclonal antibody-drug conjugates against pancreatic cancer model. Cancer Cell International, 22, 417.
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8

BiFC Plasmid Vector Screening Protocol

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The BiFC plasmid vector used for cDNA library screening was synthesized by Genscript (Piscataway, NJ, USA). Briefly, a plasmid was constructed using the pcDNA3.1+ backbone. The complementary VC155 BiFC fragment was separated from an EcoRI restriction site by a 15 amino acid serine-glycine-glycine repeat flexible linker. To validate the function of the BiFC plasmids, AC5-interacting partners were cloned into the EcoRI restriction site, and the orientation was validated by sequencing. CAD VN-AC5/D2L cells were plated in 6-well dish and cultured to 80–90% confluency, the culture media was replaced, and cells were transfected with BiFC plasmid controls using Lipofectamine 2000 according to the manufacturer’s recommendations. Images were collected 48 h after transfection using a Nikon A1 confocal microscope system and NIS-Elements 4.5 software (Nikon Instruments, Melville, NY, USA) and analyzed using ImageJ software (NIH, 1.47v, Bethesda, MD, USA). Line profile analysis was performed using NIS Elements 4.0 software (Nikon Instruments, Melville, NY, USA).
Doyle T.B., Muntean B.S., Ejendal K.F., Hayes M.P., Soto-Velasquez M., Martemyanov K.A., Dessauer C.W., Hu C.D, & Watts V.J. (2019). Identification of Novel Adenylyl Cyclase 5 (AC5) Signaling Networks in D1 and D2 Medium Spiny Neurons using Bimolecular Fluorescence Complementation Screening. Cells, 8(11), 1468.
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9

Quantification of Germinal Centers and T Follicular Helper Cells

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GCs and GC-resident TFH cells were detected using immunofluorescence staining. Briefly, 6-μm sections were cut and adhered to silanized slides and stained with the following antibodies: mouse anti-human CD4 (1F6, Leica Microsystems Inc., Buffalo Grove, IL), goat anti-human PD-1 (AF1086, R&D Systems, Minneapolis, MN), or goat anti-human CD20 (MS4A1, Origene, Rockville, MD) and rabbit anti-human Ki67 (SP6, ThermoFisher Scientific). Sections on slides were pretreated in 1 mM EDTA (pH 8.0) in a Presto pressure cooker (National Presto Industries, Eau Claire, WI) at 121°C for 35 seconds to unmask antigens. Sections were blocked with normal horse serum, and incubated with primary antibodies overnight at 4 °C. After washing in PBS, the slides were incubated at room temperature for 2 h with donkey AlexaFluor 647 anti-mouse IgG, AlexaFluor 488 donkey anti-goat IgG and AlexaFluor 594 donkey anti-rabbit IgG and counterstained with DAPI (4',6-diamidino-2-phenylindole), all from Life Technologies (Carlsbad, CA). After washing in PBS, the slides were coverslipped and examined using a Nikon A1 confocal microscope system (Nikon, Melville, NY). GCs were enumerated and size quantified using ScanScope slide scanner and ImageScope software (Aperio-Leica Biosystems, Buffalo Grove, IL).
Vargas-Inchaustegui D.A., Demers A., Shaw J.M., Kang G., Ball D., Tuero I., Musich T., Mohanram V., Demberg T., Karpova T.S., Li Q, & Robert-Guroff M. (2016). Vaccine-induction of LN-resident SIV Env-specific TFH cells in rhesus macaques. Journal of immunology (Baltimore, Md. : 1950), 196(4), 1700-1710.
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10

Immunofluorescence Staining Protocol

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Immunofluorescence was performed as previously described.7 (link)13 (link)14 (link) Simply, for immunofluorescence, the samples were prepared as smears. The smears were permeabilized with 0.2% Triton X-100 for 30 min, blocked with 30% donkey serum for 1 h, and incubated with ACTL7A (1:50; HPA021624, Atlas Antibodies, Stockholm, Sweden) at 4°C overnight. On the 2nd day, the slides incubated with donkey anti-rabbit conjugated with Alexa Fluor 488 (1:300; A32731, Invitrogen, Carlsbad, CA, USA) for 1 h at room temperature and observed under Nikon A1+ Confocal Microscope System (Nikon, Tokyo, Japan).
Yang T.Y., Chen Y., Chen G.W., Sun Y.S., Li Z.C., Shen X.R., Zhang Y.N., He W., Zhou D., Shi H.J., Xin A.J, & Sun X.X. (2022). Sperm-specific protein ACTL7A as a biomarker for fertilization outcomes of assisted reproductive technology. Asian Journal of Andrology, 24(3), 260-265.
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