Imaging of pollen hydration on stigmas, hydration in a humid chamber and GUS histochemical staining (Methods S1) of flowers and leaves was carried out using a Nikon (Surrey, UK) SMZ1500 dissection microscope coupled to Nikon Digital Sight DS‐U1 camera. A Nikon Eclipse 90i epifluorescence microscope (×10 objective) with Nikon Digital Sight DS‐U1 camera was used for imaging pollen tubes stained with aniline blue and for anthers stained for GUS activity.
Digital sight ds u1 camera
Manufactured by Nikon
Sourced in Japan, United States
The Digital Sight DS-U1 camera is a digital camera designed for microscope imaging. It features a high-resolution CMOS sensor and is capable of capturing images and video at various resolutions. The camera connects to a computer via a USB interface for image acquisition and processing.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse 80i, by Nikon (4 mentions)
Smz1500 microscope, by Nikon (3 mentions)
Tritc conjugated phalloidin, by Merck Group (2 mentions)
Nis elements ar 2, by Nikon (2 mentions)
Eclipse 90i microscope, by Nikon (2 mentions)
Smz 1500 dissection microscope, by Nikon (1 mentions)
E600 microscope, by Nikon (1 mentions)
Durcopan, by Merck Group (1 mentions)
Hoechst 33258, by Merck Group (1 mentions)
Plan fluor 40x, by Nikon (1 mentions)
Glycerol, by Merck Group (1 mentions)
Eclipse te2000 u microscope, by Nikon (1 mentions)
Cell culture incubator, by Thermo Fisher Scientific (1 mentions)
Eclipse ts100 microscope, by Nikon (1 mentions)
80i microscope, by Nikon (1 mentions)
Mantra quantitative pathology workstation, by PerkinElmer (1 mentions)
Eclipse te2000 u, by Nikon (1 mentions)
Ultra cruz mounting medium containing dapi, by Santa Cruz Biotechnology (1 mentions)
Triton x 100 pbs, by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
21 protocols using digital sight ds u1 camera
1
Imaging of Pollen Hydration and GUS Activity
2
Histological Analysis of Dorsal Root Regeneration
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At 12–16 weeks post-injury terminally anesthetized animals underwent transcardial perfusion with heparinized 0.9% NaCl solution followed by 4% paraformaldehyde in 0.1 M phosphate buffer (PB). The L5 spinal cord segment and the replanted dorsal root were rapidly removed and post fixed with 4% paraformaldehyde in 0.1 M PB for 2 h at room temperature. For spinal cord sections, the spinal cord tissue was embedded in paraffin wax and cut in 25 µm transverse sections using a microtome. For semi-thin reimplanted dorsal root sections, tissue was osmicated and then dehydrated in a graded series of ethanol and then embedded in Durcopan (Fluka, Sigma-Aldrich GmbH). After polymerization, the specimens were cut in 0.5 µm semi-thin sections on a LKB Ultrotome V and stained with toluidine blue. Images were obtained with a NIKON E600 microscope with a 60 × oil immersion plan-apo lens and a Nikon Digital Sight DS-U1 camera or a NIKON EZC1 confocal imaging system.
3
Nuclear Morphology Assessment of S2 Cells
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S2 cells (negative control), S2 cells stably expressing TnBVank1 and H. virescens haemocytes isolated 56 h after injection of the vector pIZT/V5-His with or without TnBVank1 were stained with Hoechst 33258 (Sigma-Aldrich, St Louis, MO, USA) to detect nuclear morphology. Cells were fixed with 4% paraformaldehyde for 10 min, washed with PBS, and stained with 10 μg/mL Hoechst 33258 at room temperature for 10 min in the dark. The cells were washed with PBS 1X three times and the slides were mounted with glycerol for morphologic observation by fluorescence microscopy (NIKON Eclipse 80i) equipped with a Nikon Plan Fluor 40x and 100x/0.5–1.3 Oil Iris objective and the images were recorded with Nikon Digital Sight DS-U1 camera.
4
Immunofluorescent Labeling of S2 Cells
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For the immunofluorescent labeling, S2 cells were fixed for 10 min at room temperature in 4% formaldehyde in 1X phosphate buffer saline (PBS) pH 7.5. After permeabilisation for 30 min in 0.1% Triton X-100 solution and washing in PBS 1X, the cells were incubated in 1% BSA for 30 min and, subsequently, with the mixture of two primary antibodies, anti-TnBVANK145 (link) and anti-Alix53 (link), 1:200 in BSA 1%, in a humidified chamber at 4 °C overnight. After three washes in PBS 1X, the cells were incubated with the mixture of two secondary antibodies, which were raised in different species (with two different fluorochromes, i.e. TRITC-conjugated against mouse and FITC-conjugated against rabbit) (Sigma-Aldrich St Louis, MO, USA), 1:1000 in 1% BSA for 1 h at room temperature in dark condition. After washing in PBS 1X, the slides were mounted with FluoroshieldTM with DAPI, histology mounting medium (Sigma-Aldrich St Louis, MO, USA).
To measure the total cell fluorescence, the slides were observed microscopically with Nikon Eclipse 80i equipped with a Nikon Plan Fluor 100x/0.5–1.3 Oil Iris objective and the images were recorded with Nikon Digital Sight DS-U1 camera. The intensity of the cell fluorescence was measured using the ImageJ software (http://rsbweb.nih.gov/ij/ ).
To measure the total cell fluorescence, the slides were observed microscopically with Nikon Eclipse 80i equipped with a Nikon Plan Fluor 100x/0.5–1.3 Oil Iris objective and the images were recorded with Nikon Digital Sight DS-U1 camera. The intensity of the cell fluorescence was measured using the ImageJ software (
5
Visualizing Haemocyte Actin Cytoskeleton
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Haemocytes were isolated 56 h after injection of the vector pIZT/V5-His with or without TnBVank1 as described above and from parasitised larvae. Cells were fixed, permeabilised and blocked as previously described. Subsequently, haemocytes were incubated with TRITC-conjugated phalloidin (Sigma-Aldrich St Louis, MO, USA), diluted 50 µg/ml in 1% BSA for 2 h at room temperature in dark condition. After washing in PBS 1X three times, the slides were mounted with glycerol (Sigma-Aldrich St Louis, MO, USA).
The slides were observed microscopically with Nikon Eclipse 80i equipped with a Nikon Plan Fluor 100x/0.5–1.3 Oil Iris objective and the images were recorded with Nikon Digital Sight DS-U1 camera.
The slides were observed microscopically with Nikon Eclipse 80i equipped with a Nikon Plan Fluor 100x/0.5–1.3 Oil Iris objective and the images were recorded with Nikon Digital Sight DS-U1 camera.
6
Microscopic Analysis of Cell Morphology
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The morphology of Vero and NIH3T3 cells grown in the presence of extracts of
composite was observed by microscopy (34 (link)). The morphology and relative density
of Vero cells cultured in the presence of 100-extract or supplemented DMEM were
examined using SEM (JEOL JSM 6480 Brand LV; Iniqui, Argentina). Cells were
plated at a density of 10,000 cells/cm2 on glass coverslips, and
incubated in the presence of 100-extracts or supplemented DMEM for 4 days. After
incubation, samples were washed with PBS and fixed overnight in 4%
glutaraldehyde buffered in PBS at 4°C. Next, the samples were washed 3 times
with PBS and dehydrated in graded ethanol (10%-100%) for 10 minutes each step.
Finally, after critical point drying, the samples were mounted on stubs and were
sputtered coated with gold.
NIH3T3 cell morphology was analyzed by light microscopy. NIH3T3 cells were
suspended in 0.5 mL of supplemented DMEM and seeded in 24-well plates (10,000
cells/well). After incubation for 24 hours, the media were removed, and
100-extract from each composite was added. The culturing was stopped at days 2
and 5 of culture, and cell images were captured digitally (Eclipse TE2000-U
microscope, Digital Sight DS-U1 camera, Nikon).
composite was observed by microscopy (34 (link)). The morphology and relative density
of Vero cells cultured in the presence of 100-extract or supplemented DMEM were
examined using SEM (JEOL JSM 6480 Brand LV; Iniqui, Argentina). Cells were
plated at a density of 10,000 cells/cm2 on glass coverslips, and
incubated in the presence of 100-extracts or supplemented DMEM for 4 days. After
incubation, samples were washed with PBS and fixed overnight in 4%
glutaraldehyde buffered in PBS at 4°C. Next, the samples were washed 3 times
with PBS and dehydrated in graded ethanol (10%-100%) for 10 minutes each step.
Finally, after critical point drying, the samples were mounted on stubs and were
sputtered coated with gold.
NIH3T3 cell morphology was analyzed by light microscopy. NIH3T3 cells were
suspended in 0.5 mL of supplemented DMEM and seeded in 24-well plates (10,000
cells/well). After incubation for 24 hours, the media were removed, and
100-extract from each composite was added. The culturing was stopped at days 2
and 5 of culture, and cell images were captured digitally (Eclipse TE2000-U
microscope, Digital Sight DS-U1 camera, Nikon).
7
In vivo Zebrafish T-ALL Cell Transplantation Assay
8
MRC5 Fibroblast Culture Protocol
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Human MRC5 fetal lung fibroblasts (ATCC® CCL-171™) that are between passage 20 and 30 were cultured in Roswell Park Memorial Institute (RPMI 1640) supplemented with 10% heat-inactivated fetal bovine serum (FBS), together with 100 μg/mL streptomycin and 100 units/mL penicillin. The cells were maintained in a cell culture incubator (Thermo Electron Corporation, Madison, WI, USA) with humidified atmosphere of 37°C and 5% CO2:95% O2, before passaging. The morphology of cells was examined under the Nikon Eclipse TS100 microscope (Nikon Corporation, Tokyo, Japan) attached with a Digital Sight DS-U1 camera. The setup was also equipped with the ACT-2U imaging software version 1.60.
9
Microscopic Imaging and Analysis
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Observations, measurements, line illustrations, and LM photomicrographs were made using a Nikon Eclipse 80i (Nikon, Tokio, Japan) microscope with differential interference contrast (DIC) optics, a drawing tube (camera lucida) and a Nikon Digital Sight DS-U1 camera. Photographs were edited using Adobe® Photoshop® CS software.
10
AqNOS Gene Expression in Sponge Larvae
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AqNOS gene expression patterns were assessed in precompetent (3 hpe) and competent (5 hpe) A. queenslandica larvae (Supplementary Fig. 3 ). Fixation and storage of specimens, and whole mount in situ hybridisation (WMISH) were performed as described in Larroux et al.93 94 (link). 798 and 1649 bp fragments of the AqNOS cDNA were used as probes (Supplementary Fig. 4 ). WMISH images were captured with Olympus BX60 using a Nikon Digital Sight DS-U1 camera.
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