Mitochondrial membrane potential (ΔΨm) was performed by JC-1 assay according to the previous report (Zhong et al., 2012a (link)). Briefly, cells were cultured in 6-well plates at a density of 2 × 105/well and in black 96-well plates (with transparent bottom) at a density of 2 × 104/well, respectively. The incubation periods were performed as required. Then the culture medium was removed and the cells were washed twice with PBS. A further incubation with loading dye buffer (containing 2.5 μg/mL JC-1 and 10 mM glucose) was performed for 15 min at 37°C. The cells were harvested and mean fluorescence intensity of FITC was detected using a flow cytometry (BD FACS CantoTM, BD Biosciences, San Jose, CA, United States). Furthermore, JC-1 fluorescence was also observed via fluorescent microscopy and the images were captured using an Axiovert 200 fluorescent microscope (Carl Zeiss) and AxioCam HRC CCD camera (Carl Zeiss).
Axiocam hrc ccd camera
Manufactured by Zeiss
Sourced in Germany, France
The AxioCam HRc is a CCD camera from Zeiss designed for microscopy applications. It features a high-resolution sensor and is capable of capturing high-quality images. The core function of the AxioCam HRc is to provide researchers and scientists with a reliable and efficient tool for image acquisition and analysis in their microscopy workflows.
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Lab products found in correlation
Axioplan 2 imaging microscope, by Zeiss (7 mentions)
Axioplan 2 fluorescence microscope, by Zeiss (5 mentions)
Ncl 4e2, by Abcam (3 mentions)
Matrigel, by BD (3 mentions)
Axiovision 4, by Zeiss (2 mentions)
Crystal violet, by Merck Group (2 mentions)
Axio vert a1 microscope, by Zeiss (2 mentions)
Axiovision software, by Zeiss (2 mentions)
Imager m2 microscope, by Zeiss (2 mentions)
Mz16f, by Leica (2 mentions)
Axiocam erc5s camera, by Zeiss (2 mentions)
Axio imager a1 microscope, by Zeiss (2 mentions)
Axiophot microscope, by Zeiss (2 mentions)
Facscanto, by BD (1 mentions)
Axiovert 200 fluorescent microscope, by Zeiss (1 mentions)
Ec plan neofluar objective, by Zeiss (1 mentions)
Npm fc 61991, by Thermo Fisher Scientific (1 mentions)
Alexa 488 or 594, by Thermo Fisher Scientific (1 mentions)
Ab36865, by Abcam (1 mentions)
Ab12286, by Abcam (1 mentions)
42 protocols using axiocam hrc ccd camera
1
Mitochondrial Membrane Potential Measurement
2
Immunostaining Protocol for Cellular Imaging
3
Root Length and Meristem Analysis
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Root length and meristem length were assessed in at least five independent experiments. Primary root length was measured using the software analysis package Fiji. For statistical, we used R software, we applied one-way ANOVA and Tukey’s multiple comparison test for the experiments.
To measure number of cortex cells, meristem length and cell size, seedlings were fixed in Hoyer’s solution for 30 min. The material was observed on a Zeiss Axioplan imaging 2 microscope under DIC optics. We used Poisson’s test for the experiment. Images were captured on an Axiocam HRC CCD camera (Zeiss) using the Axiovision program (version 4.2). Images analysis was performed using software package Fiji. The size of elongated cells were measured on two cells per root, on cells immediately before to the first root hair cell on six individuals per treatment.
To measure number of cortex cells, meristem length and cell size, seedlings were fixed in Hoyer’s solution for 30 min. The material was observed on a Zeiss Axioplan imaging 2 microscope under DIC optics. We used Poisson’s test for the experiment. Images were captured on an Axiocam HRC CCD camera (Zeiss) using the Axiovision program (version 4.2). Images analysis was performed using software package Fiji. The size of elongated cells were measured on two cells per root, on cells immediately before to the first root hair cell on six individuals per treatment.
4
Transwell Invasion Assay with Metformin and Cisplatin
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The upper chamber of a Transwell insert (8-μm pore size) was coated with 100 μL of Matrigel (BD Biosciences, Bedford, MA, USA) and PBS, followed by drying for 30 min at 37 °C. Cells were suspended in serum-free medium (100 μL; 4 × 105 cells/mL) and layered in the upper compartment of the chamber. The bottom chambers were supplemented with 500 μL of complete medium (10% FBS) containing the indicated concentrations of both metformin and cisplatin. After incubation for 24 h, the invading cells on the lower face were fixed in 4% paraformaldehyde and stained with crystal violet (Sigma-Aldrich). Random fields were counted, and representative images were obtained using an AxioCam HRC CCD camera (Carl Zeiss, Oberkochen, Germany).
5
Fluorescent Protein Expression in Yeast
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Yeast strains transformed with TF-P2A-GFP expressing plasmids were grown exponentially in liquid YPD media containing 200 mg/l G418 sulfate and were washed three times in PBS buffer (136.89 mM NaCl, 8.09 mM Na2HPO4, 1.76 mM KH2PO4, 2.68 mM KCl) before microscopy. Then the cells were viewed by both bright-field and fluorescence microscopy using an Axio Vert.A1 microscope (Carl Zeiss, Göttingen, Germany) equipped with an AxioCam HRc CCD camera (Carl Zeiss, Germany).
6
Cell Invasion Assay using Matrigel
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The upper chamber of a transwell insert (8 μm in pore size) was coated with 100 μl of a 1:6 mixture of Matrigel (BD Biosciences, Bedford, MA) and PBS (invasion assay), and dried for 30 min at 37 °C. Cell suspension (100 μl; 4× 105 cells/ml) in serum-free medium was placed in the upper compartment of the chamber. The bottom chambers were supplemented with 500 μl complete medium (10% FBS) containing the indicated concentrations of resistin. After incubation for 24 h, the non-migrant cells from the upper face were scraped using a cotton swab. The invaded cells on the lower face were fixed with 4% paraformaldehyde (PFA), and stained with crystal violet (Sigma-Aldrich, St. Louis, MO, USA). Random fields were counted, and representative images were photographed using the AxioCam HRC CCD camera (Carl Zeiss).
7
Immunofluorescence Analysis of Nucleolar Proteins
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HeLa cells grown on glass coverslips were fixed with 3.7% paraformadehyde and permeabilized with 0.5% NP40. The following primary antibodies were used: Rabbit anti-FBL (ab5821, Abcam), rabbit anti-Drosha (ab12286, Abcam) and rabbit anti-DGCR8 (ab36865, Abcam). Antibodies were detected with secondary antibodies conjugated to Alexa 488 or 594 (Molecular Probes, Invitrogen) and nuclei were counterstained with Hoechst 33258. Images were captured using Axioplan2 fluorescence microscope (Zeiss) equipped with AxioCam HRc CCD-camera and AxioVision 4.5 software using EC Plan-Neofluar 40x/0.75 objective (Zeiss). Image quantification was carried out according to refs. [34] , [35] (link).
8
Tracing Glucose Intake in Ants
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To confirm that the glucose water diet did not spread over the ant cuticle, a 20% glucose solution labelled with non-toxic fluorescent green drain tracing dye (Hydra) was fed to ants. Five mg ml−1 of dye was added to a 20% glucose solution, of which 300 μl was supplied to ants in the cap of an Eppendorf. Ants were sampled just after taking a feed, and after 6 and 24 h of being exposed to the dye, to trace the spread of the solution over time. After sampling, ants were carefully fixed on their backs, and brightfield and fluorescent images were acquired using a Zeiss M2 Bio Quad SV11 stereomicroscope (Additional File 1 : Fig. S2). The samples were illuminated either with a halogen lamp (brightfield) or a 100-W Hg arc lamp (fluorescence) and reflected-light images were captured with an AxioCam HRc CCD camera and AxioVision software (Carl Zeiss, Cambridge, UK). Green fluorescence was excited with light passed through a 470-nm filter (40 nm bandpass), and the emission was collected through a 525-nm filter (50 nm bandpass).
9
Mitochondrial Membrane Potential Assay for Apoptosis
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The mitochondrial membrane potential (MMP) assay of intact cells was stained with JC-1 before visual determination [19 (link)]. JC-1 probe is a dual-emission fluorescent dye that can reflect the changes in MMP. It forms aggregates that result in a red emission in normal polarized mitochondria, while it forms monomers that emit green fluorescence on the depolarized mitochondrial membrane, the MMP depolarized is an early phenomenon of apoptosis [20 (link)]. MCF-7 and MDA-MB-231 cells were seeded onto 96-well plates. After a 24 h incubation for adhesion, the cells were treated with the indicated compounds for 4 h. The medium was removed and the cells were incubated with JC-1 probe for 30 min. Then, the medium with the probe was removed and the cells were rinsed with phosphate-buffered saline (PBS). The cells were observed with a fluorescent microscope, and images were obtained with an Axiovert 200 fluorescent inverted microscope (Carl Zeiss) and an AxioCam HRc CCD camera (Carl Zeiss).
10
Carpel Development Imaging Protocol
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Flowers from different developmental stages with at least 40 ovules per pistil were dissected and cleared for 2hrs in Hoyers solution. For GUS staining, developing carpels and siliques were dissected and incubated in GUS staining buffer (5 mM EDTA, 0.1% Triton X-100, 5 mM K4Fe(CN)6, 0.5 mM K3Fe (CN)6, and 1 mg mL–1 X-Gluc [Rose Scientific] in 50 mM sodium phosphate buffer, pH 7.0) for 24 – 48hrs at 37°C. The dissected pistils were observed on a Zeiss Axioplan imaging 2 microscope under DIC optics. Images were captured on an Axiocam HRC CCD camera (Zeiss) using the Axiovision program (version 4.2).
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