A1r gaasp confocal microscope

Manufactured by Nikon

The A1R GaAsP confocal microscope is a high-performance imaging system designed for advanced microscopy applications. It utilizes a Gallium Arsenide Phosphide (GaAsP) detector to provide enhanced sensitivity and low-light detection capabilities. The A1R GaAsP is capable of performing confocal imaging, allowing for optical sectioning and high-resolution imaging of samples.

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Lab products found in correlation

Alexa 488 conjugated phalloidin, by Thermo Fisher Scientific (1 mentions) A11004, by Thermo Fisher Scientific (1 mentions) Prolong gold dapi, by Thermo Fisher Scientific (1 mentions) Imaris 6, by Oxford Instruments (1 mentions) Ab6556, by Abcam (1 mentions)

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A1R confocal microscope (1179 citations) Confocal microscope (523 citations) A1R microscope (119 citations) A1R confocal (60 citations) A1R GaAsP Inverted Confocal Microscope (6 citations) A1R GaAsP (3 citations)

4 protocols using a1r gaasp confocal microscope

Visualizing Phosphoinositide Dynamics in Drosophila Wings

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Wings were dissected from w[1118] (FlyBase: FBal0018186) pupae and aged for 28 h after prepupa formation. These were fixed for 30 min in 4% paraformaldehyde/0.2% glutaraldehyde in PBS at room temperature, the outer cuticle was removed, and then quenched for 15 min in 50 mM NH₄Cl in PBS.
Subsequent staining steps were carried out on ice or at 4°C. Wings were blocked and permeabilized for 45 min in 5% BSA/0.5% Saponin in PIPES buffer. Recombinant biosensors were diluted in 0.1% Saponin in PIPES buffer (PI(4)P biosensor at a final concentration of 150 nM and PI(4,5)P₂ biosensor at 500 nM) with 1:200 Alexa488-conjugated Phalloidin (Invitrogen) and incubated with the wings for 1 h. Wings were then washed 12 times in 0.1% Saponin in PIPES buffer, post-fixed for 10 min in 2% PFA, washed a further four times with 0.1% Saponin in PIPES buffer, and mounted in 2.5% DABCO with 10% glycerol in PBS. The wings were imaged on a Nikon A1R GaAsP confocal microscope using a 60× NA1.4 apochromatic lens within 1 week of mounting. Images were acquired posterior to longitudinal vein 4 with a pixel size of 110 nm and z-sections spaced by 150 nm.

Maib H., Adarska P., Hunton R., Vines J.H., Strutt D., Bottanelli F, & Murray D.H. (2024). Recombinant biosensors for multiplex and super-resolution imaging of phosphoinositides. The Journal of Cell Biology, 223(6), e202310095.

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Quantification of DNA Damage Response Proteins

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1–5 × 10⁵ cells were plated onto poly-d-lysine treated coverslips and allowed to attach prior to adenovirus infection and/or drug treatment. To stain, coverslips were fixed in 4% paraformaldehyde, antigens retrieved with a 0.2% Triton x-100 PBS solution, and cells blocked with PBS plus 5% normal goat serum and 0.3% Triton X-100. Antibody dilutions are as follows: BRCA1 (sc-6954), γH2AX 1:1000 (05-3636, Millipore), RPA2 1:300 (ab2175), RAD51 1:500 (sc-8349), goat anti-mouse Alexa Fluor 568 1:500 (A-11004, ThermoFischer Scientific, Waltham, MA), and goat anti-rabbit Alexa Fluor 633 1:500 (A-21070). Nuclei were counterstained with DAPI ProLong Gold (P-36931, Life Technologies). RPA and RAD51 foci were captured on a Carl Zeiss Apotome instrument, and foci within 100 cells counted per time point per replicate. BRCA1, RAD51, and RPA foci counts were performed by eye due to background signal from the antibodies. High resolution images were imaged on an inverted Nikon A1R GaAsP confocal microscope. Whole cell γH2AX foci were also collected on the confocal microscope using z-stacks and were quantified using Imaris 6 (Bitplane AG, Zurich, Switzerland).

Smith E.A., Gole B., Willis N.A., Soria R., Starnes L.M., Krumpelbeck E.F., Jegga A.G., Ali A.M., Guo H., Meetei A.R., Andreassen P.R., Kappes F., Vinnedge L.M., Daniel J.A., Scully R., Wiesmüller L, & Wells S.I. (2017). DEK is required for homologous recombination repair of DNA breaks. Scientific Reports, 7, 44662.

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Immunocytochemistry Protocol for Cell Imaging

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For immunocytochemistry, gels were rinsed 2x with PBS prior to fixation with 4% paraformaldehyde (30 minutes). Gels were rinsed 3x with PBS and then permeabilized with blocking buffer (BB – 2% normal goat serum, 0.2% Triton-X) for 30 minutes. Primary antibodies (tyrosine hydroxylase, β3-tubulin, human nuclear antibody, MAP2, DAT and Nurr1) were incubated at 4° C overnight prior to washing with PBS and incubation with the appropriate secondary antibodies for 3 hours. Samples were washed to remove background stain and fixed with Ibidi mounting media prior to imaging on a Nikon A1R Ga-Asp confocal microscope. Image quantification was performed using ImageJ. 3D stacks were compressed into a single image using maximal z-projection. Manual background subtraction was performed after measuring the background fluorescence of each gel where no cell was identified. Cells were then manually counted.

Godbe J.M., Freeman R., Burbulla L.F., Lewis J., Krainc D, & Stupp S.I. (2020). Gelator length precisely tunes supramolecular hydrogel stiffness and neuronal phenotype in 3D culture. ACS biomaterials science & engineering, 6(2), 1196-1207.

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Drosophila Pupal Wing Polarity Analysis

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Adult wings were dehydrated in isopropanol and mounted in GMM (50% methyl salicylate, 50% Canada Balsam).
Unless otherwise indicated, pupal wings were dissected at 28 hr after puparium formation (APF) at 25°C, as previously described [50] . Primary antibodies for immunostaining were mouse monoclonal anti-Fmi (Flamingo #74, DSHB) [58] , rat anti-Dsh [24] , affinity purified rabbit anti-Fz [59] , rabbit anti-Stbm [15] , affinity purified rat anti-Pk [18] , affinity purified rabbit anti-GFP (ab6556, Abcam), mouse monoclonal anti-β gal (DSHB), rabbit anti-β gal (#55976, Cappel), rat monoclonal anti-Ecad (DSHB) and mouse monoclonal anti-Arm N2 7A1 (DSHB).
Actin was labelled with Alexa568-conjugated Phalloidin (A12380, Molecular Probes).
Pupal wings were imaged on a Nikon A1R GaAsP confocal microscope using a 60x NA1.4 apochromatic lens. 9 Z slices separated by 150 nm were imaged, and then the 3 brightest slices around cell junctions were selected and averaged for each channel in ImageJ.
Membrane masks were generated in Packing Analyzer [60] . Polarity magnitude (maximum asymmetry ratio on a cell-by-cell basis) and the variation in polarity angle were calculated as previously described [49] . Values were compared using one-way ANOVA with Dunnett's multiple comparisons test, or using paired t-tests for clones, comparing control and mutant regions of the same wings.

Strutt H., & Strutt D. (2020). DAnkrd49 and Bdbt act via Casein Kinase Iε to regulate planar polarity inDrosophila.

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