For the metaphase assay and immunofluorescence, we performed the imaging on a microscope (Carl Zeiss, imager.M2) fitted with a 63× oil-immersion objective lens and 10× ocular lens. We processed and analysed the images with ZEN 2.3 imaging software (Carl Zeiss Microscopy GmbH, Jena, Germany). For the Boyden chamber and wound-healing assays, we imaged the pictures using a microscope (Carl Zeiss, Vert.A1) fitted with a 10× or 5× objective lens and 10× ocular lens. We analysed the images using ZEN 2.3 imaging software and ImageJ software (ImageJ 1.51f, Wayne Rasband, National Institutes of Health, USA). For the gelatine matrix degradation assays, we performed the imaging on a microscope (Carl Zeiss, imager.M2) fitted with a 40× objective lens and 10× ocular lens. We processed and analysed the images using ZEN 2.3 imaging software and ImageJ software.
Imager m2
Manufactured by Zeiss
Sourced in Germany, United States, Belgium
The Imager M2 is a high-performance microscope imaging system designed for advanced materials science applications. It provides high-resolution imaging capabilities, enabling precise analysis and characterization of a wide range of samples.
Automatically generated - may contain errors
Lab products found in correlation
Apotome 2, by Zeiss (15 mentions)
Axiocam 506, by Zeiss (9 mentions)
Lsm 880, by Zeiss (8 mentions)
Zen software, by Zeiss (7 mentions)
Axiovision software, by Zeiss (7 mentions)
Axiocam mrc5, by Zeiss (4 mentions)
Vectashield, by Vector Laboratories (4 mentions)
Axio imager, by Zeiss (4 mentions)
Antibody diluent, by Agilent Technologies (4 mentions)
Axiocam erc5, by Zeiss (3 mentions)
Apotome 2 module, by Zeiss (3 mentions)
Zen v2.3 blue edition software, by Zeiss (3 mentions)
Horse serum, by Vector Laboratories (3 mentions)
Novared, by Vector Laboratories (3 mentions)
Duolink in situ pla, by Merck Group (3 mentions)
Axio scan z1, by Zeiss (3 mentions)
Apotome module, by Zeiss (3 mentions)
Vectashield with dapi, by Vector Laboratories (3 mentions)
Dylight 488 or 594 conjugated secondary antibody, by Thermo Fisher Scientific (3 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (2 mentions)
184 protocols using imager m2
1
Microscopy Imaging and Analysis Protocols
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Visualizing Cellulose and Callose in Plant Roots
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To stain cellulose, Mitra and Loque [61 ] protocol for Calcofluor staining was followed with some modifications. Briefly, the roots were transferred to 2 ml tubes and stained with 0.02% calcoflour white (Sigma) for 5 min. The primary root was placed on the microscope glass slide and transverse section of middle part of the roots was prepared using sharp blade. Calcofluor White was visualized using an epifluorescence microscope (Zeiss Imager M2).
The staining of callose was done according to Muller et al. [62 (link)] with some modifications. Briefly, the roots were stained with 0.1% (w/v) aniline blue solution in 0.1 M sodium phosphate buffer (pH = 7.2) for 1.5 h. The cross-section of roots was done as described above and visualized under epifluorescence microscope (Zeiss Imager M2).
The staining of callose was done according to Muller et al. [62 (link)] with some modifications. Briefly, the roots were stained with 0.1% (w/v) aniline blue solution in 0.1 M sodium phosphate buffer (pH = 7.2) for 1.5 h. The cross-section of roots was done as described above and visualized under epifluorescence microscope (Zeiss Imager M2).
3
Quantitative Fluorescent Imaging of RPE/Choroidal Flat Mounts
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ISH of RPE/choroidal flat mounts was performed using the Multiplex Fluorescent Reagent Kit v2 (ACD) according to the protocol of Gross-Thebing et al. [26 (link)]. The following probes were used: Iba1 (channel C1 or C3), VEGF (channel C3), and PGF (channel C1). Images were taken with a Zeiss Imager M.2 including an ApoTome.2.
The colored pixel intensity in individual image areas of the laser spots was quantified using the Colored Pixel Counter tool for Fiji.
The colored pixel intensity in individual image areas of the laser spots was quantified using the Colored Pixel Counter tool for Fiji.
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Adipocyte Size Analysis via Microscopy
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Samples were analyzed by microscope Zeiss Imager.M2 using a 10× objective. The size of 200 cells was measured for each animal using ImageJ (Rasband, W.S., ImageJ, U. S. National Institutes of Health, Bethesda, Maryland, United States, https://imagej.nih.gov/ij/ , 1997–2018) by manually drawing the outline of each adipocyte.
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Histological Analysis of Gibel Carp Gonads
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The gonads of gibel carp were fixed with 4% paraformaldehyde in PBS at 4°C over night. After washing with PBS, the samples were immersed in 30% saccharose-PBS buffer for 5 h at 4°C, embedded in paraffin, and then were cut into 4μm sections. Hematoxylin-eosin staining was performed as described previously [50 (link)]. Immunofluorescence co-localization of Gsdf and Vasa was performed as described previously [71 (link)]. The images were obtained by upright fluorescence microscope Axio Imager M2 (Carl Zeiss).
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Quantifying Parvalbumin and Perineuronal Nets
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Brains were sectioned in a cryostat at 35 μm and prepared for fluorescent immunohistochemistry as previously described (Xu, Ouyang, Xiong, Stary, & Giffard, 2015 ). Briefly, sections were treated with sheep anti‐parvalbumin (PVALB; [1:500]; R&D Signaling cat# AF5058) and the PNN‐binding fluorescent wisteria floribunda agglutinin (WFA; [1:500]; Vector Labs cat# FL‐1351) and incubated in 594 nm secondary (Invitrogen cat# A‐11016). Z‐Stacks (1μm thick X 35 sections) were acquired through cornu ammonis‐1 (CA1), cornu ammonis‐3 (CA3), and somatosensory cortex using a Zeiss Imager M2 equipped with an Apotome 2 for optical sectioning and collapsed into a maximum‐projection image for cell quantification. Average PVALB+ and PNN+ cell counts were acquired from six hippocampal sections per animal, using maximum projection images by an individual blinded to experimental group. Neocortical cell counts were acquired from the same sections in the adjacent somatosensory/barrel cortex, dorsolateral to the rostral hippocampus, in order to limit variability in PVALB+ and PNN+ counts that could be due to tissue processing.
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Cell Morphology Evaluation of Leukemia Cells
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After the appropriate time of Ro treatment (or DMSO in controls) in culture, 1.5 × 105 MLL-AF9 BM or human leukemia cells (K562 and MOLM13) were washed once with 1× PBS, counted and centrifuged onto slides for 5 minutes at 35 × g and air-dried for 24 h prior to Richard-Allan Scientific Three-Step Stain Staining Set (Thermo Scientific) based on Eosin Y and Methylene Blue/ Azure A and mounted with Permount solution (Fisher). Cell morphology was evaluated by light microscopy at ×400 magnification (Zeiss Imager M-2 equipped with AxioCam ERc 5 s).
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Immunofluorescence Staining of Cells
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Cells were fixed for 20 min at 4°C in 4% paraformaldehyde and washed in PBS. Cells were incubated for 20 min at room temperature in phosphate buffered saline Triton (PBST) (0.1% Triton X-100; Sigma; in PBS) containing 10% donkey serum (Serotec Ltd.) and incubated overnight at 4°C with primary antibody diluted in 1% donkey serum in PBST. Cells were then washed in PBS and incubated with secondary antibodies for 2 h at room temperature. Hoechst 33258 was used to visualize DNA. See Supplementary Table S1 (Supplementary Data are available online at www.liebertpub.com/scd ) for a complete list of primary antibodies used in this study. Samples were imaged using a Zeiss Imager M.2, equipped with AxioCam MRm and MRc cameras and AxioVision software for image capture and scale bars=100 μM.
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Detecting EBNA1-NCL Protein Interactions
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Cells were fixed in 1× PBS, 4% paraformaldehyde for 20 min and permeabilized for 10 min with 0.4% Triton X-100, 0.05% CHAPS. The EBNA1–digoxigenin probe mRNA (5′-CTTTCCAAACCACCCTCCTTTTTTGCGCCTGCCTCCATCAAAAA-3′) at 50 ng/well was denaturated for 5 min at 80°C. The probe hybridization reaction was carried out in 40 μl of hybridization buffer (10% formamide, 2× SCC, 0.2 mg/ml E. coli tRNA, 0.2 mg/ml salmon sperm DNA and 2 mg/ml BSA). The fixed cells were washed and blocked into the blocking solution (1× PBS, 3% BSA, 0.1% saponin) before incubation with the primary antibodies (anti-digoxigenin, Sigma 1/200 and anti-NCL, Abcam 1/1000). The PLA reaction was performed under the manufacturer’s protocol using the Duolink PLA in situ kit, PLA probe anti-rabbit plus, PLA probe anti-mouse Minus and the in situ detection reagent FarRed, all from Sigma. The results were analysed using a Zeiss Axio Imager M2. All the PLA experiments were performed at least three times independently, and the following controls probes were implemented: without mRNA probe or without primary antibodies.
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Immunofluorescent Labeling of Inner Ear Sensory Organs
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