Seven DIV neurons expressed NgCAM-Halo for 12 h and treated with 500 nM JF646 (Gross et al., 2013 (link)) to bind all of the folded NgCAM-Halo proteins. Cells were washed twice with conditioned N2 medium and incubated with medium containing DMSO or 10 μM propofol. Cover slips were fixed with 4% paraformaldehyde at 1, 2, 4, 8, and 12 h after the JF646 washout. Fifteen minutes before fixation, neurons were treated with 500 nM JF549. Control cells (0 min) were not exposed to JF646 dye but incubated with JF549 at 12 h and fixed. Cover slips were imaged with an Axio Imager Z1 with a plan-apochromat 63× 1.4 NA objective and an Axiocam 506 mono EMCCD camera. Image analysis was performed in ImageJ/Fiji and Excel. A region of interest was drawn around the Golgi and two to three axon terminals for each cell, and the 20% brightest pixels were identified and their mean intensity determined.
Axiocam 506 mono
Manufactured by Zeiss
Sourced in Germany
The Axiocam 506 mono is a monochrome digital camera designed for microscopy applications. It features a 6.5-megapixel CMOS sensor and supports high-resolution image capture. The camera is capable of delivering fast frame rates and offers a wide dynamic range for accurate imaging of samples.
Automatically generated - may contain errors
Lab products found in correlation
Imager z2, by Zeiss (6 mentions)
Observer z1, by Zeiss (3 mentions)
Zen software, by Zeiss (3 mentions)
Zen blue software, by Zeiss (3 mentions)
Axio zoom v16, by Zeiss (3 mentions)
Axiozoom, by Zeiss (2 mentions)
Axio imager z1, by Zeiss (2 mentions)
Hxp 120v fluorescent lamp, by Zeiss (2 mentions)
Plan apochromat, by Zeiss (2 mentions)
Dfc340 fx camera, by Leica (2 mentions)
M165 fc, by Leica (2 mentions)
Axio imager z2, by Zeiss (2 mentions)
Anodisc, by Cytiva (2 mentions)
Whatman, by Cytiva (2 mentions)
Axio imager m2, by Zeiss (2 mentions)
Axio imager z1 microscope, by Zeiss (2 mentions)
Zyla 4, by Oxford Instruments (2 mentions)
Axio imager m2 upright microscope, by Zeiss (2 mentions)
Dapi fluoromount g, by Southern Biotech (2 mentions)
Anti iba1 antibody, by Fujifilm (2 mentions)
62 protocols using axiocam 506 mono
1
Imaging Folded NgCAM-Halo Proteins
2
Microscopic Imaging of Constipated Worms
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DIC images of the constipated worms in Figure 2D were taken using a Plan Apochromat 40x/1.4 Oil DIC objective in a Zeiss Imager Z2 microscope equipped with an Axiocam 506 Mono camera using ZEN Blue 2.3 software.
3
Live-cell Migration Imaging Protocol
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Single cell migration analysis was performed as previously described (Ioannou et al., 2015) (link). The live images were obtained using a fluorescence microscope with a 37°C incubation chamber (Zeiss Observer.Z1) and equipped with a CCD camera (Axiocam 506 mono). The images were captured for the period of 11 h at 20 min intervals. The cell migration distance and the migration velocity were tracked using Fiji (ImageJ) software.
4
Quantifying Molecular Delivery Dynamics
5
H2O2-Induced Mitochondrial Dynamics in KGN Cells
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KGN cells were incubated with 100 nM MitoTracker Green FM (Molecular Probes, Eugene, OR, USA) for 30 min at 37 °C and 5% CO2 in microscopy-optimized cell culture dishes (µ-dish 35 mm, ibidi). The staining solution was prepared in DMEM/Ham’s F12 without phenol red. To examine changes of the mitochondrial structure, cells were treated with 100 µM H2O2 after staining and washing. Fluorescence was recorded with a wide-field microscope (microscope: Axio Observer.Z1; light-source: Colibri.2; camera: Axiocam 506 mono; objective: Plan-Apochromat 63x/1.40 Oil Ph 3 M27; software: ZEN 2.6; Carl Zeiss Microscopy). A 450–490 nm BP (excitation) and a 500–550 nm BP (emission) were used (F46-002; AHF analysentechnik). In a second approach, stimulation with H2O2 for 4 h in colorless medium prior to staining and imaging was performed to rule out phototoxicity due to multiple imaging as a reason for mitochondrial fragmentation.
To evaluate the effect of H2O2 (100 µM) on mitochondria, 239 control cells and 122 treated cells in two dishes each for both groups were analyzed after 4 h of treatment. Examples for KGN cells with elongatedor fragmented mitochondrial networks are given in the corresponding figure.
To evaluate the effect of H2O2 (100 µM) on mitochondria, 239 control cells and 122 treated cells in two dishes each for both groups were analyzed after 4 h of treatment. Examples for KGN cells with elongatedor fragmented mitochondrial networks are given in the corresponding figure.
6
Fluorescence Microscopy of GFP-Labeled Cells
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YAA3 cells were treated with 3.6 µM of 11β,13-dihydrolactucin or 12.5 µM of FK506 for 90 min, followed by incubation with or without 3 mM MnCl2 for further 90 min. On the last 10 min of incubation, 10 µg/mL of Hoechst 33,342 (Sigma) nuclear dye were added. Cells were washed and resuspended in 5 µL of 1,4-Diazabicyclo[2.2.2]octane (DABCO, triethylenediamine) DABCO solution (200 mM DABCO in 75% (v/v) glycerol, 25% (v/v) PBS) (Sigma-Aldrich). The preparations were monitored for GFP fluorescence as previously described [27 (link)] using a Zeiss Imager Z2 (Zeiss, Germany) fluorescence microscope. Photographs were taken with an Axiocam 506 mono (Zeiss). Three images were taken and analyzed for each sample, each one containing ≈ 600 individual cells. Images were analyzed using Fiji-ImageJ 1.53f (NIH, Bethesda, MD, USA).
7
Imaging and Analysis of Flat-Mounted Embryos
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For imaging of flat-mounted embryos after in situ hybridisation, an AxioZoom V16 stereomicroscope (Zeiss) equipped with an Axiocam 506-Mono and a colour digital camera were used. Immunostained embryos were imaged with Zeiss LSM 800 or 880 with Airyscan confocal microscopes. For live imaging, embryos were aligned on heptane glue coated coverslips and submersed in a thin layer of halocarbon oil. Bright-field live imaging was performed using an AxioZoom V16 stereomicroscope, while fluorescence live imaging was performed with confocal microscopes. Image stacks were processed in Fiji (Schindelin et al., 2012 (link)) and Helicon Focus (HeliconSoft). Image brightness and intensity was adjusted in Corel PhotoPaint X5 (CorelDraw) and Fiji.
8
Visualizing OATP2B1 and MRP1 Expression
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Cells were seeded in Ibidi 4 chambers pH+ µ-slide (Gräfelfing, Germany) 48 hours before microscopic analysis using Zeiss Observer Z1 (Zeiss, Germany). The fluorescence signals were obtained using a Zeiss Colibri 7 LED light source. Red fluorescence (OFPSpark) emitted by the tag attached to OATP2B1 was observed with a yellow LED at an excitation wavelength of 555 nm and an emission quadrature bandpass 582/30. EGFP emitted by the tag attached to MRP1 was observed with a blue LED at an excitation wavelength of 475 nm and an emission quadrature bandpass 514/30. Images were captured with a Zeiss Axiocam 506 mono and processed using Zeiss Zen 2 lite.
9
Visualizing Protein Aggregation in Worms
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Whole-worm images of day 4 adults (Q34), L4s (Q40), or day 1 adults (α-synuclein) were taken with an Axiocam 506 mono (Zeiss) camera using the 5×, 10×, or 20× objective of a Zeiss Axio Imager Z1 microscope at defined exposure time (Q35: 20 ms, Q40: 10 ms, α-synuclein: 10 ms). Aggregate numbers were evaluated from the photos. Due to the smaller size and relatively lower number of aggregates in the α-synuclein model, a maximum intensity projection was created from a Z-stack of the head region and aggregates present from the nose tip to the posterior bulb were counted. Genotypes of the samples were blinded during the counting. Aggregates were defined as discrete structures or puncta above background.
10
Multimodal Imaging of Cellular Structures
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Device imaging was performed using inverted fluorescent microscopy (Olympus IX83 microscope, Olympus; Tokyo, Japan) with MetaMorph software (Molecular Devices; San Jose, CA), with the following exceptions: Growth factor titration, KI-67, CC3, and UEA-1 experiments were imaged using a Zeiss Axio Observer.Z1 microscope with a Zeiss Axiocam 506 mono camera (Carl Zeiss AG; Oberkochen, Germany). The integrated GOC model was imaged using a Zeiss LSM 880 II Airyscan FAST confocal microscope with the accompanying Zeiss Zen Microscope software, as was the EC only bead perfusion study. DAPI staining of HIEC monolayers on Transwells was imaged using a Leica SP8X tandem scanning confocal microscope with a white light laser 40x oil objective, with LASX by Leica Microsystems software (Leica Microsystems; Wetzlar, Germany).
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