Growth factor reduced (GFR) Matrigel (120 μl per chamber; Corning, 356231) was added to 8-chamber culture slides, incubated at 37 °C for 20 min and washed with PBS. ECWT/WT were pretreated with conditioned medium from LFBWT/WT or LFBΔ/Δ for 24 h, trypsinized and seeded on Matrigel-coated slides at a density of 2 × 104 cells per chamber. Cells were imaged at 4 h after plating using an Axiovert 40CFL inverted microscope (Zeiss) equipped with AxioCam MRM CCD camera (Zeiss).
Axiocam mrm ccd camera
Manufactured by Zeiss
Sourced in Germany
The AxioCam MRm is a CCD camera from Zeiss, designed for microscopy applications. It features a high-resolution sensor and supports a variety of image acquisition modes. The camera is intended to capture images and data from microscopic samples.
Automatically generated - may contain errors
Lab products found in correlation
Axiovision software, by Zeiss (17 mentions)
Axio observer z1, by Zeiss (6 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (6 mentions)
Axio observer z1 microscope, by Zeiss (5 mentions)
Axiovision 4, by Zeiss (5 mentions)
Apotome, by Zeiss (4 mentions)
Turbofect, by Thermo Fisher Scientific (4 mentions)
Axio imager epifluorescence microscope, by Zeiss (4 mentions)
Poly d lysine coated culture slides, by BD (3 mentions)
Axioimager m2 microscope system, by Zeiss (3 mentions)
Plan apochromat 63 na 1.40 objective, by Zeiss (3 mentions)
Anti phospho histone h3, by Merck Group (3 mentions)
Vectashield mounting medium with 4 6 diamidino 2 phenylindole, by Vector Laboratories (3 mentions)
Axiovision rel 4, by Zeiss (3 mentions)
Alexa fluor 594, by Thermo Fisher Scientific (3 mentions)
Cellobserver z 1, by Yokogawa (3 mentions)
Csu x1, by Zeiss (3 mentions)
Vectashild, by Vector Laboratories (2 mentions)
Axioimager m1 fluorescent microscope, by Zeiss (2 mentions)
A11008, by Thermo Fisher Scientific (2 mentions)
78 protocols using axiocam mrm ccd camera
1
Angiogenic Potential of Endothelial Cells
2
Alternaria alternata Genetic Manipulation
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The wild-type EV-MIL31 strain of Alternaria alternata (Fr.) Keissler was isolated from diseased citrus leaves [5 (link)] and used for transformation and mutagenesis. Fungal strains defective for a SKN7 response regulator (Δskn7), a high osmolarity glycerol (Δhog1) mitogen-activated protein kinase, and a “two component” histidine kinase (Δhsk1) were created from the EV-MIL31 strain in previous studies [7 (link),8 (link)]. Unless otherwise stated all fungal strains were grown on potato dextrose agar (PDA) (Difco, Sparks, MD) at 28°C. Fungal mycelium was ground in sterile water using a disposable pestle (Fisher Scientific, Atlanta, GA), evenly spread onto a layer of cellophane overlaid on PDA for 3 to 5 days, and harvested for DNA or RNA isolation. For conidia formation, fungal strains were grown on PDA without parafilm sealing in light for 5 days, and harvested by immersing, scraping with sterile water, passing through Miracloth and centrifugation (5,000 xg). Conidia were examined with a Zeiss Axio Imager A1 microscope equipped with an AxioCam MRm CCD camera (Oberkochen, Germany). Germination of conidia was assessed by placing them on glass slides or 96-well microtiter plates and incubating in a plastic box for 6 h. Fungal hyphae were treated with cell wall degrading enzymes to release protoplasts in a solution containing sucrose as an osmotic agent as described [35 (link)].
3
FISH Karyotyping of Cannabis sativa
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The following probes were used: pTa71 (18S-28S rDNA) [11 (link)], pCT4.2 (5S rDNA) [69 (link)], the CS-1 probe (C. sativa subtelomeric repeat JX402748), and the CS-237 probe (C. sativa repeat ON055366). CS-1 (a clone preserved from the research by Divashuk et al., 2014), 5S rDNA, and 18S-28S rDNA were labeled by nick-translation with digoxigenin-11-dUTP, and CS-237 was labeled by PCR with biotin-16-dUTP according to the manufacturer’s instructions (Boehringer, Ingelheim am Rhein, Germany). The FISH experiments were performed as described by Karlov et al. [70 (link)]. The stringency of the FISH was about 72% (washing conditions: 15 min in 0.1× SSC at 42 °C). The chromosomes were counterstained with 1 mg/mL DAPI and mounted in Vectashild (Vector Laboratories, UK). An AxioImager M1 fluorescent microscope (Zeiss, Oberkochen, Germany) was used to observe the chromosome preparations. The metaphase plates with fluorescent signals were photographed with a monochrome AxioCam MRm CCD camera and visualized using the Axiovision software (Zeiss). The metaphase chromosomes were classified according to Levan et al. [71 (link)] based on their arm ratios and FISH hybridization patterns.
4
Time-lapse Microscopy of Live Cells
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Time-lapse recordings with phase contrast or double fluorescence applications were performed on a Zeiss Axio Observer Z1 inverted microscope with Zeiss Plan Neofluar 10x, 0.3 NA objective coupled to a Zeiss Axiocam MRM CCD camera and equipped with a Marzhauser SCAN-IM powered stage. Zeiss Colibri LED illumination system was used for fluorescent excitation. For structured illumination microscopy and fluorescent optical sectioning we used Zeiss Apotome module. During time-lapse imaging the cell cultures were kept in a stage-top incubator (CellMovie) providing for required temperature and CO2 atmosphere. Power stage positioning, illumination, focusing, optical sectioning and primary image collection were controlled by Zeiss Axiovision 4.8 software and a custom-made experiment manager software module on a PC. Images were further processed using Zeiss Axiovision 4.8 and NIH ImageJ software.
5
Quantifying CFTR in B. cenocepacia-infected Macrophages
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One million macrophages were cultured on 12 mm glass cover slips in 24-well tissue culture plates and infected synchronously with B. cenocepacia at an MOI of 2 due to clumping at higher MOIs. Macrophage nuclei were stained blue with DAPI. CFTR was detected with CFTR antibody (2784, Abcam, CF3) followed by fluorescent secondary antibodies (Molecular Probes, A11008). Microscopy was performed using an Axiovert 200 M inverted epifluorescence microscope equipped with the Apotome attachment for improved fluorescence resolution and an Axiocam MRM CCD camera (Carl Zeiss Inc., Thornwood, NY). Five independent fields with at least 10 macrophages were scored for each condition. All experiments were performed in triplicate.
6
Immunofluorescence Staining of mCCDcl1 Cells
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mCCDcl1 cells were seeded on to Corning Costar™ Transwell™ Permeable Supports (0.4 μm pore size) in 24 well plates at 1×105 cells/cm2 in complete growth medium. After 7–9 days, cell monolayers were washed in PBS for two times and fixed in 100% methanol (chilled at -20°C) for 5 min at RT, followed by three times wash in ice cold PBS for 5 min each as done before [27 (link), 34 (link)]. The cells were then blocked in blocking buffer (PBS, 10% goat serum and 0.05% Triton X-100) for one hour. Whereupon primary antibodies were add to the blocking buffer at 1:100 dilution and incubated overnight at 4°C. The following primary antibodies were used: anti-SK1 (Alomone, APC-039), anti-SK3-ATTO-594 (Alomone, APC-025-AR), anti-IK1 (Alomone, ALM-051), anti-BKα (Alomone, APC-021) and anti-TRPV4 (Alomone, ACC-034). After primary antibody incubation, the cells were washed 3 times in PBS, 5 min each, followed by incubation with secondary antibody (Life Technologies, alexa fluor 488 or 594 labeled goat anti-rabbit) in blocking buffer. The cells were then washed another three times in PBS, 5 min each, and mounted with ProLong Gold Antifade Mountant with DAPI (Life Technologies). Cells were imaged at 100x using a Nikon A1R Confocal Laser Microscope or a Zeiss Axioskop 40 microscope equiped with a AxioCam MRm CCD camera.
7
Immunofluorescence Assay for Transfected Cells
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For IFAs of transfected BHK-21 cells or intracellular parasites grown in host cells (HFF), cells were fixed with 4% formaldehyde in PBS and permeabilized with 0.1% Triton X-100 in PBS/3% BSA. Coverslips were blocked in PBS 10% FCS and proceeded further for IFA as previously described [26 (link)]. Samples were observed with a Zeiss Axioimager epifluorescence microscope equipped with an apotome and a Zeiss Axiocam MRmCCD camera driven by the Axiovision software (Zeiss), at the Montpellier RIO imaging facility. Images were collected and processed using Zeiss Zen software.
8
Quantifying Tumor Proliferation and Angiogenesis
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Frozen tissue sections were fixed in methanol/acetone and blocked in 12% bovine serum albumin (BSA)/PBS at RT for 30 min prior to the application of the primary rabbit polyclonal antibody against Ki67 (abcam; 1:200) and the rat monoclonal antibody against CD31 (BD Pharmingen, Heidelberg, Germany; 1:200). Sections were then incubated with a secondary anti-rabbit Alexa488-conjugated antibody (Jackson ImmunoResearch, West Grove, Pennsylvania, USA) for Ki67 staining and secondary anti-rat Cy-3-conjugated antibody (Jackson ImmunoResearch) for CD31 staining along with Hoechst bisbenzimide (5 μg/ml) to counterstain nuclei. Sections were embedded using Fluorescent Mounting Medium (Dako, Hamburg, Germany) and imaged at 10× magnification on an Axiovert 135 TV fluorescence microscope (Carl Zeiss, Munich, Germany) equipped with an AxioCam MRm CCD camera (Carl Zeiss) and AxioVision Rel. 4.8 software (Carl Zeiss). Immunostainings were captured at identical illumination conditions, exposure time and system settings for digital image processing. Quantification of cellular proliferation (percentage of Ki67 positive cells in the tumor) and blood vessel density (percentage of CD31 positive area in the tumor) was performed by evaluation of 4–5 visual fields per tumor using ImageJ software (NIH).
9
Comprehensive Imaging of C. elegans Gonad
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Whole-worm DAPI staining was carried out as described (Angelo and Van Gilst 2009 (link)). SYTO-12 (Life Technologies) staining for apoptotic cells was performed as described (Gumienny et al. 1999 (link)). Antibody staining of dissected gonads was carried out as described (Kershner et al. 2014 (link)). Primary antibodies were used at the following dilutions: rabbit anti-GLP-1 (1:20) (Crittenden et al. 1994 (link)), rabbit anti-REC-8 (1:500; SDIX), rabbit anti-HIM-3 (1:200) (Zetka et al. 1999 (link)), mouse anti-DAO-5 (1:200; Developmental Studies Hybridoma Bank), and mouse anti-MPK-1(YT) (1:400; Sigma). The secondary antibodies Alexa fluor 488 goat anti-rabbit and goat anti-mouse (Molecular Probes) were used at a dilution of 1:2000 and 1:1000, respectively. Images were captured with Nomarski optics using a Zeiss Axioplan2 microscope and a Zeiss AxioCam MRm CCD camera. Identical microscope and camera settings were used to acquire images for both the control and experimental groups. Images were processed by Adobe Photoshop, and the signal intensity was quantified using ImageJ.
10
Immunocytochemistry of TSPO in Cells
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C20 wildtype or TSPO knockdown/knockout cells were grown for 24 h on sterile glass coverslips and fixed for 10 min at room temperature with 4% (w/v) paraformaldehyde (Carl Roth GmbH, Karlsruhe, Germany). After washing, cells were permeabilized with blocking/permeabilization solution (10% (v/v) goat serum, 0.5% (v/v) Triton X-100 in 1 × PBS) for 20 min. Cells were then incubated overnight, at 4 °C with rabbit-anti-TSPO antibody (ab109497) and mouse-anti-ATPB antibody (ab14730), both from Abcam, Cambridge, UK, diluted 1:1000 in 2% goat serum and 0.1% Triton X-100 in 1 × PBS. After three additional washing steps, cells were incubated for 1 h with secondary antibodies conjugated with Alexa Fluor 488 and Cy3 (Life Technologies, Carlsbad, CA, USA, both diluted 1:1000 in 2% goat serum and 0.1% Triton X-100 in 1 × PBS. Nuclei were labeled with Hoe33342 (AppliChem, Darmstadt, Germany) at a final concentration of 0.1 ug/mL in 1 x PBS. Finally, cells were mounted with confocal matrix (Micro Tech Lab, Graz, Austria) and examined with an inverted fluorescence microscope (Observer.Z1, ZEISS, Jena, Germany). An XBO 175 W served as the light source (Lambda DG4, Sutter instruments, Novato, CA, USA). Images were taken by a ZEISS AxioCam MRm CCD camera using the ZEN software (ZEISS).
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