Cells were permeabilized by incubation with 0.5% Triton X-100 at 4°C for 10 min, washed three times with 1x PBS-T (1x PBS+0.05% Tween 20) and blocked using 10% BSA solution (dilution with PBS-T) at room temperature for 1 hour. The samples were stained with a primary antibodies: Hsp90 (Cell signaling technology), phosphorylated HSF-1Ser326 (Novus Biological) and phosphorylated HckTyr410 (Thermofisher Scientific). For nuclear staining, the samples were counter stained with 4′,6-diamidino-2-phenylindole (DAPI; Thermo Fisher Scientific). Images were taken on a Nikon Eclipse Ti camera (Nikon Instruments) with NIS Elements Imaging Software (3.10). Confocal fluorescence imaging was performed on Zeiss LSM 800, Airyscan Confocal Laser Scanner Microscope with Zen 2.3 software. Post processing of the images was completed either in Image J or Zen lite software.
Eclipse ti camera
Manufactured by Nikon
Sourced in Japan
The Nikon Eclipse Ti camera is a high-performance microscope imaging system. It features a cooled scientific-grade CMOS sensor and advanced optics to capture high-quality, low-noise images. The camera is designed for use in a wide range of laboratory applications, including cell biology, materials science, and other fields requiring detailed microscopic imaging.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Hsp90, by Cell Signaling Technology (1 mentions)
Nis elements imaging software, by Nikon (1 mentions)
Nis elements microscope imaging software, by Nikon (1 mentions)
Nunc lab tek 2 chambered coverglass, by Thermo Fisher Scientific (1 mentions)
Oct compound, by Sakura Finetek (1 mentions)
Calcein, by Thermo Fisher Scientific (1 mentions)
Leitz dm il microscope, by Leica (1 mentions)
Plan apo vc 20x dic n2, by Nikon (1 mentions)
10 protocols using eclipse ti camera
1
Immunofluorescence Analysis of Cellular Signaling
2
Alkaline Phosphatase Staining Protocol
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Alkaline phosphatase staining was performed using the Alkaline Phosphatase Staining Kit II (Stemgent) according to the manufacturer’s instructions. The cells were photographed using a Nikon Eclipse Ti camera (Nikon).
3
Imaging and Quantification of Nanoscale Connections
4
Fluorescence Imaging and Quantification
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Fluorescence imaging was performed on a Nikon Eclipse Ti camera (Nikon Instruments) with NIS-Elements Microscope Imaging Software (3.10). Confocal fluorescence imaging was done on a PerkinElmer Ultraview Spinning Disk Confocal Microscope with Velocity acquisition software and Hammamatsu ORCA-ER CCD camera. Contrast and brightness parameter adjustments were applied across the whole image or equally across all the comparison groups when necessary using NIS-Elements Microscope Imaging Software (3.10). The length, width, and nodal area of the vessel structures was measured in more than 200 images, as well as the elongation of more than 300 epithelial cells aligned along the endothelial vessel structures, using NIS-Elements software. Z-stack images were processed using the NIS-Elements Advanced Research deconvolution module to generate 3D reconstruction images.
5
Characterization of mitotic defects
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HeLa cells were transfected with siControl or sicGAS and then were treated with nocodazole (100 ng/mL) for 16 h. Then, cells at prometaphase were collected by shaking the dish, and then they were seeded (1 × 104 cells/well) in a four-well glass dish (Thermo Scientific™ Nunc™Lab-Tek II Chambered Coverglass, MA, USA) and incubated overnight in standard culture conditions to enable estimation of the duration of mitosis along with visualization of unstable chromosomal phenotypes with time-lapse photomicroscopy. To visualize chromosomes, cells were incubated with 1 μg/mL Hoechst 33342 (Thermo Scientific™ Hoechst® 33342, MA, USA) for 30 min. Fluorescence images were acquired every 5 min for 24 h while using a Nikon eclipse Ti camera (Tokyo, Japan) with a ×40 dry Plan-Apochromat objective. Images were captured with an iXonEM +897 Electron Multiplying charge-coupled device camera (Teledyne Princeton Instruments, Trenton, NJ, USA) and analyzed in the Nikon Imaging Software (NIS)-elements advanced research (AR) (Nikon, Tokyo, Japan).
6
Imaging and Quantification of Nanoscale Connections
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Fluorescence imaging was performed on a Nikon Eclipse Ti camera (Nikon Instruments) with NIS Elements Imaging Software (3.10). Confocal fluorescence imaging was done on a PerkinElmer Ultraview Spinning Disk Confocal Microscope with Velocity acquisition software and Hammamatsu ORCA-ER CCD camera. Contrast and brightness parameter adjustments were applied across the whole image or equally across all the comparison groups when necessary. Quantification was done using NIS Elements Software; we measured the length of the complete and broken nanoscale connections, as well as other projections. Z-stack images were processed using the deconvolution software, to generate 3D reconstruction images.
7
Histological Evaluation of Aortic Samples
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Histological evaluation was performed following the method described in previous studies.9 (link),18 (link), 19 (link), 20 (link) Samples of the aorta from each group were fixed overnight in 4% formalin. Afterward, formalin was substituted sequentially with 10%, 20%, and 30% sucrose solution at intervals of at least 10 hours. Then, the samples were embedded in O.C.T. compound (Sakura Finetek Japan, Tokyo, Japan) and frozen in liquid nitrogen. The samples were stored at ˗80°C until analysis. Frozen sections (7 µm thick) from each group were adhered to charged slides and air-dried overnight. The sections were then stained with Masson's trichrome as previously described,9 (link),18 (link), 19 (link), 20 (link) microscopically examined, and photographed using an Eclipse Ti camera (Nikon, Tokyo, Japan).
8
Platelet Adhesion Assay in BOEC Microfluidics
9
Quantifying Wound Healing Histology
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After the fixation of the wound, this was included in paraffin. For each wound included, three sections were obtained with 5 µm thickness in a rotating microtome (Microm / HM-315). One of the cuts was stained with 0.25% Toluidine Blue in MacIlvaine pH 4.0 buffer, another with Gomori Trichrome and last with Sirius Red. These stains were used for the quantification of mast cells, blood vessels and collagen, respectively. For all analyzes, for each slide, ten photomicrographs of different areas were obtained. For the quantification of mast cells and blood vessels, the images were obtained in the Leica Microsystems Inc. type microscope (Wetzlar) with the 40x objective (400x of magnitude) in a coupled camera. For the differentiation and quantification of collagen types I and III, these photomicrographs were obtained with 20x objective (200X of magnitude), captured in the Nikon eclipse Ti camera, optic camera with polarization filter. All quantifications were performed with the Imagem software (Rasband 2011) . For the quantification of mast cells and blood vessels, we used the multiple point tool and for the quantification of collagen, we used the threshold tool. The values were represented by the average as described below:
1 wound per animal = 1 histological slide = 10 different areas Average= total quantification results obtained in the 10 areas / 10 (number of areas evaluated)
1 wound per animal = 1 histological slide = 10 different areas Average= total quantification results obtained in the 10 areas / 10 (number of areas evaluated)
10
Stomatal Characteristics of Arabidopsis Rosette Leaves
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The experiments were performed using fully developed rosette leaves of 35-day-old Arabidopsis plants. The cleared epidermal peels from abaxial leaf surfaces and nail polish immersions from adaxial leaf surfaces were prepared and examined with a light microscope equipped with a Nikon Eclipse Ti camera, DS-Fi1c-U2 optics and Plan Apo VC 20x DIC N2.
The counts were made on 6 leaves from independent plants for each ecotype and then averaged.
For the measurements of stomatal aperture preparations were made from epidermal peels from abaxial leaf surfaces. Before preparation the leaves were incubated in a buffer composed of 10 mM KCl, 0.1 mM EGTA and 10 mM MES-KOH at pH 6.15 for 2 hours. Measurements were analysed using a light microscope, Eclipse Ti camera, DS-Fi1c-U2 optics and Plan Apo VC 20x DIC N2 (Nikon,Tokyo, Japan). The ratio of the length and width of the pore between guard cells was calculated using the NIS-Element program. The counts were made on 6 leaves from independent plants.
The counts were made on 6 leaves from independent plants for each ecotype and then averaged.
For the measurements of stomatal aperture preparations were made from epidermal peels from abaxial leaf surfaces. Before preparation the leaves were incubated in a buffer composed of 10 mM KCl, 0.1 mM EGTA and 10 mM MES-KOH at pH 6.15 for 2 hours. Measurements were analysed using a light microscope, Eclipse Ti camera, DS-Fi1c-U2 optics and Plan Apo VC 20x DIC N2 (Nikon,Tokyo, Japan). The ratio of the length and width of the pore between guard cells was calculated using the NIS-Element program. The counts were made on 6 leaves from independent plants.
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