To image cells, between 0.05 and 0.1 g of sediment (estimated) was placed in a sterile 2 mL tube, and 0.1 mL of sterile phosphate buffered saline (PBS) pH 7.4 was added. The tube was vortexed to mix the sediment, and 30 μL of the suspended sediment/PBS mixture was transferred to a clean tube. 3 μL of a 1:100 dilution of 1 mg mL−1 DAPI (4, 6-diamidino-2-phenylindole) stock solution was added, and the sediment suspension was stained for 10 min in the dark. The suspension was then centrifuged at 7600 g for 2 min, and the fluid removed. Fifteen microliters of sterile PBS pH 7.4 was then added to the tube and mixed. This suspension was observed on a Zeiss Axio Observer DI and photographed with an attached Zeiss MRc camera (Zeiss, Oberkochen, Germany) and Axiovision software (Zeiss). Images were adjusted for contrast and brightness using GraphicConverter.
Mrc camera
Manufactured by Zeiss
Sourced in Germany
The Mrc camera is a high-performance digital camera designed for microscopy applications. It features a large sensor size and high resolution to capture detailed images with excellent image quality. The camera is compatible with a wide range of microscopes and can be used for various imaging techniques, including brightfield, fluorescence, and phase contrast.
Automatically generated - may contain errors
Lab products found in correlation
Axiovision software, by Zeiss (1 mentions)
Axio observer di, by Zeiss (1 mentions)
Axio imager z1 microscope, by Zeiss (1 mentions)
Apotome structured illumination, by Zeiss (1 mentions)
Dmrb microscope, by Zeiss (1 mentions)
Bh 2 light microscope, by Olympus (1 mentions)
Axioskop microscope, by Zeiss (1 mentions)
Axioskop, by Olympus (1 mentions)
Autoquant x3, by Media Cybernetics (1 mentions)
Volocity 3d image analysis software, by PerkinElmer (1 mentions)
Matrigel, by BD (1 mentions)
Binocular loop, by Leica (1 mentions)
Two chamber transwell assay, by Corning (1 mentions)
Axio observer z1, by Zeiss (1 mentions)
Siport neofx transfection agent, by Thermo Fisher Scientific (1 mentions)
48 well microchemotaxis chamber, by Neuro Probe (1 mentions)
Transwell insert, by Corning (1 mentions)
Axiovert microscope, by Zeiss (1 mentions)
Axiocam hrc, by Zeiss (1 mentions)
Microtome, by Leica (1 mentions)
13 protocols using mrc camera
1
DAPI Staining of Sediment Microbes
2
Multimodal Imaging of Biological Samples
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Immunofluoresence images were captured with a Zeiss Axio Imager Z1 microscope and Zeiss MRm camera, processed and false-colored using Zeiss Axiovision software. High magnification images were captured using Zeiss Apotome structured illumination. Histology (hematoxylin and eosin) images were captured with a Leica DMRB microscope using a Zeiss MRc camera and processed using Zeiss MRGrab software.
3
Microscopic Spore Measurement Protocol
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Specimens were mounted in 2 % KOH, Melzer’s reagent or cotton blue in lactic acid and studied with a Zeiss Axioskop or an Olympus BH2 light microscope using bright field or phase contrast optics. Spore measurements were made without ornamentation and ornament height was recorded separately. Spore measurements are given as a range covering 90 % of measured spores with 5 % extreme values given within parentheses. The number of collections used (x) and spores measured (y) is provided in the form n = x/y. Spores were photographed in a Zeiss Axioskop microscope equipped with a ×100 DIC objective lens, a Zeiss MRc camera, and Zen Blue software. Photographed tissue was mounted in Melzer′s reagent.
4
3D Imaging of Neural Structures
5
Transwell Assay for Cell Migration and Invasion
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Migration and invasion were determined by a two-chamber transwell assay (Corning Incorporated; Corning, NY, USA). The upper side of the polycarbonate film was left untreated (migration) prior to cell seeding or was covered with Matrigel™ (500 ng/μL; BD Biosciences; Franklin Lakes, NJ, USA) (invasion, chemotaxis assay). 600 μL of complete medium was added into the lower chamber. After succinate addition, 2 × 104 cells were suspended in 100 μL supplement-free medium and added into the upper chamber (inserts). After incubation for 24 h at 37 °C, transwells were gently picked up, invaded cells on the bottom of the inserts were rinsed with PBS. Nucleic acids were stained with 0.05% crystal violet and photographed with a binocular loop (Leica; Wetzlar, Germany) microscope equipped with a Mrc camera (Zeiss; Oberkochen, Germany). Cells were counted using the ImageJ software (NIH; Bethesda, MD, USA).
6
miR-106b Regulates Cell Migration
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Scratch assay was used to investigate the effects of miR-106b expression on cell migration. Briefly, 6.6 × 104 WRO and 1.9 × 105 TPC1 cells were transiently transfected with miR-106b mimic, si-C1orf24 or respective scramble negative controls using siPORT neoFX transfection agent to WRO cells according to manufacturer’s instructions (Ambion) and cell electroporation (150 V, 900 μF) to TPC1 cells. After 24 hours of transfection, the “wound gap” was introduced, by scraping the cell monolayer with a pipette tip, into WRO and TPC1 cultures. Then, the medium was replaced and incubated in a humidified chamber at 37°C with 5% CO2, integrated with Zeiss Axio Observer Z1 inverted phase microscope system and equipped with a MRc camera (Carl Zeiss, Göttingen, Germany). Images at time zero were captured to record the initial area of wounds. Subsequently, images were taken every one hour, over the course of 72 h, at 5x magnification. Cell migration toward the wounds was expressed as percentage of wound closure. Area of the wound was quantified using CorelDraw Graphics Suite X5 (Ottawa, Canada). Experiments were performed in quintuplicates.
7
Visualizing kinesin-driven microtubule dynamics
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A 1:4 ratio of TRITC-labeled bovine and unlabeled porcine tubulin (Cytoskeleton Inc., USA) at a concentration of 20 μM were used to prepare taxol stabilized MT-filaments in general tubulin buffer as described by the supplier (Cytoskeleton Inc., USA). Into a double backed tape chamber, we sequentially flowed in 4.1 μg/μl of a 67 kDa recombinant human kinesin (Cytoskeleton Inc., USA), blocking buffer (5 mg/ml Casein) and MT filaments. The chamber was then washed with a casein-containing buffer and the reaction was started with 1 mM ATP with anti-fade mix (0.05 M glucose, 1% sucrose, 0.5 mg/ml catalase, 0.5 mg/ml glucose oxidase, 0.5% beta-mercaptoethanol (Cytoskeleton Inc., USA)). Time-series images were acquired every minute for 30 minutes on an upright epifluorescence microscope with a 40x (N.A. 0.75) EC Plan Neofluar lens mounted on a Zeiss Axio Imager.Z1 (Carl Zeiss, Germany) using filters for excitation (563 nm) and emission (581 nm) and an MRC camera (Carl Zeiss, Germany).
8
Chemotaxis Assay for Cancer Cells
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A 48-well micro-chemotaxis chamber (8-μm diameter pores; Neuroprobe) and transwell inserts (Corning #353097) were used as previously described (Porporato et al., 2014 (link)), with 0.15% (SiHa-F3) or 0.5% (B16F10) FBS as a chemo-attractant. Fifty thousand SiHa-F3 cells or two hundred thousand B16F10 cells per well were allowed to migrate during 16 h. Migrated cells were then fixed with methanol, stained with Crystal Violet, and imaged with an inverted phase Axiovert microscope equipped with a Mrc camera (Zeiss). Cells were counted using the ImageJ software (NIH). Data were normalized to cell viability.
9
Whole Embryo and Sectioned Sample Imaging
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Whole embryos were imaged in 3% methylcellulose using a Zeiss Stemi SV 11 microscope with an AxioCam HRc (Zeiss) camera and AxioVision (Zeiss) software. For sectioned samples, embryos were dehydrated with an ethanol work-up and embedded in JB4 medium (Polysciences Inc.) and 7-μm-thick sections were cut with a Leica microtome. Sections were collected on glass slides and mounted with a coverslip in Aquapolymount (Polysciences Inc.) and imaged using a Zeiss Axioplan2 microscope with an MRc camera (Zeiss) and AxioVision (Zeiss) software.
10
Measuring Embryonic Tissue Dimensions
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To compare tissue dimensions in fixed embryos, we killed females and dissected embryos in phosphate-buffered saline (PBS), then fixed the embryos in phosphate-buffered 4% formaldehyde for 14–24 h at 4 °C. We stained whole embryos with 1 µg ml−1 4′,6-diamidino-2-phenylindole (DAPI) for 30 min and photographed them with a Zeiss mRc camera on a Zeiss steREO Discovery V.12 dissecting microscope that was scale calibrated. We used the linear measurement tool in Fiji/ImageJ95 (link) to measure somite and PSM lengths. We analysed these data in R (ref. 94 ) and made plots using ggplot2 (ref. 116 ).
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