Zebrafish infections were completed as previously described [25 (link),52 (link)]. The golden mutant zebrafish line and the Tg(mpeg:mCherry) red fluorescent macrophage line were used as previously described. Embryos at 30 hrs post-fertilisation were dechorionated and anaesthetised in tricaine prior to caudal vein injection with 2–3 nL of M. abscessus strains (~100 CFU/nL) expressing Tdtomato fluorescent protein under the control of pTEC27 plasmid. After infection, embryos were recovered in fresh embryo water and placed into 24-well plates with 2 embryos/well. Embryos were monitored daily, with dead embryos removed from the well daily until 12 days post-infection. At 2, 4 and 6 days post-infection, embryos were anaesthetised in tricaine and placed on 3% methylcellulose (w/v) for fluorescent microscopy using Zeiss Axio Zoom.V16 coupled with an Axiocam 503 mono (Zeiss). Granulomas were quantified based on the bacterial co-localisation with macrophage fluorescent signal. Bacterial burden was quantified in ImageJ using the ‘Analyze particles’ function to quantify bacterial fluorescence. Abscesses were quantified based on the large uncontrolled extracellular growth of fluorescent bacteria that exceeded 100 μm in diameter. All experiments were performed in at least three independent experiments.
Axiocam 503 mono
Manufactured by Zeiss
Sourced in Germany
The Axiocam 503 mono is a high-resolution microscope camera from Zeiss. It features a 5.0-megapixel CMOS sensor and supports monochrome image capture. The camera is designed for reliable and precise image acquisition in microscopy applications.
Automatically generated - may contain errors
Lab products found in correlation
Axio zoom v16, by Zeiss (6 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions)
Axio imager m2m, by Zeiss (3 mentions)
Axio imager z2, by Zeiss (3 mentions)
Zen 2, by Zeiss (3 mentions)
Apotome 2, by Zeiss (2 mentions)
Zen image acquisition software, by Zeiss (2 mentions)
Axiovert, by Zeiss (2 mentions)
Nanog, by Thermo Fisher Scientific (2 mentions)
Matrigel coated slides, by Ibidi (2 mentions)
Nanog d73g4, by Cell Signaling Technology (2 mentions)
Sox2 alexafluor488 e 4, by Santa Cruz Biotechnology (2 mentions)
Ssea4 alexafluor488, by BD (2 mentions)
Tra 160 alexafluor488, by BD (2 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (2 mentions)
Axio imager m2, by Zeiss (2 mentions)
Zen software, by Zeiss (2 mentions)
Observer z1, by Zeiss (2 mentions)
Paraformaldehyde (pfa), by Thermo Fisher Scientific (2 mentions)
Lsm 880, by Zeiss (2 mentions)
39 protocols using axiocam 503 mono
1
Zebrafish Model of Mycobacterial Infection
2
Multimodal Organelle Imaging in Yeast
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For analysis of mitochondrial morphology, yeast strains were transformed with mitochondrial targeted GFP or mCherry. For analysis of ER morphology and peroxisome accumulation, yeast strains were transformed with mCherry-Ubc6 and GFP-SKL, respectively. For analysis of vacuoles, lipid droplets and nuclei, strains were stained using FM4–64 (#T13320; Invitrogen), BODIPY 493/503 (#D3922; Invitrogen) or DAPI (#62248; Thermo Scientific), respectively, according to the manufacturer’s instructions. Exponentially growing yeast cells were analyzed by epifluorescence microscopy (Axio Imager.M2; Carl Zeiss MicroImaging, Inc.) using a 63x oil-immersion objective.38 (link) Images were acquired with a camera (Axiocam 503 mono; Carl Zeiss MicroImaging, Inc.) and processed with Zen 2 (Carl Zeiss MicroImaging, Inc.). Peroxisome and lipid droplet accumulation was quantified by measuring GFP-SKL or BODPY 491/503 fluorescence intensity, respectively, using CellProfiler 4.95 (link) Quantifications of organellar morphology are depicted as mean (bars), standard error of the mean (line) and individual replicates (circles, squares and triangles) from three independent experiments with at least 200 cells.
3
Measuring Fluorescent Intensity in S. pyogenes
4
Quantifying Circulating Tumor Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An image of the entire cell array area was obtained using a fluorescence microscope (Axio Imager M2m; Carl Zeiss, Oberkochen, Germany) integrated with a 10× objective lens and a computer-operated motorized stage, a digital camera (AxioCam 503 mono; Carl Zeiss), and ZEN image acquisition software (Carl Zeiss). We defined a CTC as a DAPI-positive, cytokeratin-positive, and CD45-negative cell. The number of CTCs in 3 mL of peripheral blood was normalized to that in 7.5 mL to compare with the CellSearch system.
5
Immunostaining for DNA Modifications
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Embryos were fixed in 4% PFA/Phosphate Buffer (PB) for 20 min at room temperature. The embryos or sections were washed in TBS with 0.5% Triton (TBS-T) and subsequently blocked with 5% FBS in TBS-T for 3 h at RT. The embryos or sections were incubated in primary antibody solution at 4°C fortwo days. The primary antibody used were mouse monoclonal anti-5mC (Abcam, ab10805, 1:500), rabbit polyclonal anti-5hmC (Active Motif, Cat Nº 39791, 1:500), goat polyclonal anti-hSOX2 (Santa Cruz Biotech; Y-17; 1:500, for Figure 2A ), rabbit monoclonal anti-SOX2 (Abcam, ab92494, 1:1,000; for Figures 6C,F ), and anti-PAX7 IgG1 (Developmental Studies Hybridoma Bank, 1:10). The secondary antibodies used were donkey anti-rabbit 594, donkey anti-mouse 488, donkey anti-goat 647, goat anti-rabbit 594, and goat anti-mouse IgG1 647 (all from Molecular Probes, 1:500). After several washes in TBS-T, the embryos and sections were mounted and imaged by using Carl ZEISS Axio observer 7 inverted microscope (Axio observer Colibri 7, Axiocam 305 color, Axiocam 503 mono) and Carl ZEISS ZEN2 (blue edition) software. The negative controls omitting the primary antibodies (anti-5mC and anti-5hmC) fails to detect any specific mark (see Supplementary Figure S1 ).
6
Hep3B Cell Culturing and Spheroid Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hep3B cells were plated at high density (500,000 cells/ml) in regular tissue culture 10 cm dish (for monolayer) or in low attachment flasks (for spheres) in complete MEM and cultured for 48hrs with a media change at 24hrs. The spheres were stained with Hoechst-33342 and imaged at 48hrs using Zeiss Observer.Z1 microscope equipped with Axiocam 503 mono (Zeiss) camera. The individual channel images of Hoechst-33342 were pseudo-colored to red, merged with bright field and exported using ZEN 2.3 Pro software (Carl Zeiss Microscopy, GmbH, 2011, Blue edition). The final composite was done using Adobe Photoshop CS5 (Adobe Systems Inc., San Jose, CA, USA). Similar experiments were performed to collect monolayer cells and spheres for RNA isolation for RT-PCR/qRTPCR analysis using Hep3B parental cells or miR-520G stably transfected cells.
7
Microtubuli Polymerization Dynamics Visualization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Microtubuli polymerization studies were performed on the Zeiss Axio Observer Spinning Disc system equipped with a Plan-Apochromat 63x/1.4 Oil DIC objective and two Axiocam 503 mono CCD cameras. Clip170-GFP and DsRed2-Paxillin were acquired sequentially every second for 2 min using an automated emission filter wheel for CSU-X1. The following excitation lasers and emission filters were used: GFP, 488 nm diode laser, 525/50 nm filter, RFP, 561 nm (RFP) diode laser, 600/50 nm filter. Kymographs were generated in the Zen blue 2.1 software.
8
Quantifying Bacterial Biofilm Composition
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To quantify the total adhered bacteria visualized after DAPI staining as well as the vital and non-intact bacterial cells (live/dead staining) on each material, 15 different locations of the initial two-hour biofilms were screened for each material sample and wearing cycle. All samples were analyzed using FM with a 63 × oil immersion objective (ApoTome.2, Zeiss, Oberkochen, Germany). The active and dead cells on the images obtained were quantified using the image analysis program ZEN 2 pro (Zeiss, Oberkochen, Germany), and the coverage rates of the live and dead bacteria were subsequently calculated from the data obtained [53 (link)]. The results were also visualized by representative microscopic images that were acquired using a 3-Megapixel Microscope camera (Axiocam 503 mono, ZEISS, Oberkochen, Germany).
9
Imaging GFP::SPT-2 Expression in L4 Worms
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the analysis of the tissue expression of GFP::SPT-2, L4 animals were mounted in M9 buffer with 0.25 mM levamisole on 2% agarose pads. Imaging was performed at 20 °C using a microscope equipped with a ×63 1.25 numerical aperture oil lens (Imager M2; Carl Zeiss) and a charge-coupled device camera (Axiocam 503 mono). Nomarski and GFP Z-stacks (2 μm per slice) were sequentially acquired using the Zeiss acquisition software (ZEN v.3.1 blue edition). The same LED intensity and acquisition time was used for all images (Nomarski (50%, 20 ms), GFP (50%, 100 ms)).
10
Fluorescent Dendriplexes Internalization Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Internalization of fluorescently labeled dendriplexes was investigated by fluorescence microscopy. Cells were incubated with BDEF33 or AE2G3 complexes with or without amiR-155-FAM and alone as stated above. Then, cells were washed twice with 2 mM EDTA solution in PBS and fixed with a mixture of ethanol:glacial acetic acid (3:1, v/v). Cells were resuspended thoroughly to avoid clumps after fixation step and placed on slides.
Then, the slides were analyzed using phase-contrast microscopy for cell cytoplasm and nuclei visualization in transmitted light. After that, slides were mounted with Pro Long Gold antifade containing DAPI (Invitrogen MP, Waltham, MA, USA) to prevent dye photo-bleaching and identify cell nuclei further.
Phase-contrast, as well as fluorescent microscopy, was performed with the Axioscope 40 fluorescence microscope (Zeiss, Germany) equipped with a high-pressure mercury lamp HBO 50W, with Zeiss interference filter sets (Set No. 49 for DAPI) and CCD-chamber AxioCam 503 mono (at 1936 × 1460 px resolution and 14-bit capacity). DAPI-stained nuclei images and FAM signals were captured separately with the software package ZEN-2012 (Zeiss, Germany) on the magnitude X1000 (Figure S1 ). Exposure time was adjusted automatically.
Then, the slides were analyzed using phase-contrast microscopy for cell cytoplasm and nuclei visualization in transmitted light. After that, slides were mounted with Pro Long Gold antifade containing DAPI (Invitrogen MP, Waltham, MA, USA) to prevent dye photo-bleaching and identify cell nuclei further.
Phase-contrast, as well as fluorescent microscopy, was performed with the Axioscope 40 fluorescence microscope (Zeiss, Germany) equipped with a high-pressure mercury lamp HBO 50W, with Zeiss interference filter sets (Set No. 49 for DAPI) and CCD-chamber AxioCam 503 mono (at 1936 × 1460 px resolution and 14-bit capacity). DAPI-stained nuclei images and FAM signals were captured separately with the software package ZEN-2012 (Zeiss, Germany) on the magnitude X1000 (
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!