Fluorescence microscopy images of plant segments expressing GFP were collected using an Olympus SZX12 microscope and photographed with a DP73 digital camera (Olympus, Japan). Expression response of the cytokinin reporter construct in roots of pTCSn1::GFP-ER transformed T0 plants was observed by incubating the plants in 15 μM BAP, and fluorescence observed before treatment and after treatment at 1 h and 24 h as in [29 (link)].
Dp73 digital camera
Manufactured by Olympus
Sourced in Japan, United States, Italy, United Kingdom, Germany
The DP73 is a digital camera designed for microscope imaging applications. It features a 17.28-megapixel sensor and supports high-resolution image capture. The camera provides real-time image preview and supports various image file formats for versatile image acquisition and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Bx43 light microscope, by Olympus (6 mentions)
Bx53 fluorescence microscope, by Olympus (6 mentions)
Bx51 microscope, by Olympus (5 mentions)
Ax70 microscope, by Olympus (4 mentions)
Cellsens imaging software, by Olympus (3 mentions)
Bx51 system microscope, by Olympus (3 mentions)
Cellsens dimension software, by Olympus (3 mentions)
Cellsens dimension 1, by Olympus (3 mentions)
Proteinase k, by Roche (2 mentions)
Dako statspin hybridizer, by Agilent Technologies (2 mentions)
Mz16 microscope, by Leica (2 mentions)
Bx60 microscope, by Olympus (2 mentions)
Dapi mounting medium, by Thermo Fisher Scientific (2 mentions)
Bx53 microscope, by Olympus (2 mentions)
Bx63 microscope, by Olympus (2 mentions)
Ix71 microscopy system, by Olympus (2 mentions)
Axiovert 100 microscope, by Zeiss (2 mentions)
Bx53 light microscope, by Olympus (2 mentions)
Ix73 inverted microscope, by Olympus (2 mentions)
Dp controller software, by Olympus (2 mentions)
92 protocols using dp73 digital camera
1
Cytokinin Response in Transgenic Plants
2
Histological Analysis of Intestinal Tissues
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Excised ileum and colon tissues were fixed in 4% paraformaldehyde and embedded in paraffin. The blocks were serially cut into 5-μm-thick sections and stained with hematoxylin and eosin (H&E). Histological images were obtained using an Olympus IX53 microscope (Tokyo, Japan), an Olympus DP73 digital camera, and Olympus cellSens imaging software.
3
Immunohistochemistry Staining Protocol
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Tumors were fixed overnight with 10% formalin and embedded in paraffin. Serial sections of 4 μm thickness were prepared from formalin-fixed and paraffin-embedded blocks collected as described above. One section was stained with H&E and others were used for immunohistochemistry for each block. Immunostaining was performed manually by a conventional method: Briefly, sections were deparaffinized in xylene and rehydrated in a sequence of descending concentrations of ethanol. Endogenous peroxidase reactivity was blocked with 3% H2O2 for 10 min. For antigen retrieval, sections were submerged in 0.1 M citrate buffer (pH6.0) or 5 mM EDTA solution (pH8.0) and microwaved (500W) continuously for 20 min in a pressure cooker. For F4/80 staining, sections were treated with 200 μg/mL proteinase K (Roche) for 5 min at room temperature. Sections were then incubated with a respective primary antibody for 1.5 h at room temperature. After washing, sections were treated with Polink-2 Plus HRP RAT or RABBIT with DAB kit (GBI, Inc., Bothell, WA, USA) according to the manufacturer’s instructions. Finally, sections were counterstained with hematoxylin, dehydrated, and mounted. Images were acquired using an Olympus BX43 light microscope connected to a DP73 digital camera (Olympus, Tokyo, Japan).
4
Measuring Fibroblast Colony Formation
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The colony-forming unit fibroblast (CFU-F) assay was performed using whole spleen stromal cells prepared from tamoxifen-treated mice (P7) as described above. Cells were plated at a clonal density (1.0 × 105/9 cm2) and, after 14 days of culture, colonies were counted under a fluorescent stereomicroscope (Olympus MVX10) with a DP73 digital camera (Olympus, Tokyo, Japan). Differentiation cultures to induce adipocytes, osteocytes and chondrocytes were carried out as previously described27 (link).
5
Sudan Black Staining of C. elegans Lipids
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Sudan Black (Sigma-Aldrich, St Louis, MO, USA) staining of stored fat was performed as previously described45 (link),46 (link) with slight modification. A piece of frozen culture containing space-flown C. elegans was slowly thawed in an equal amount of 2% paraformaldehyde in M9 buffer, and then fixed for 30 min at room temperature. The fixed worms were then subjected to three freeze–thaw cycles, and then sequentially dehydrated through washes with 25, 50, and 70% ethanol. Staining was performed overnight in a saturated Sudan Black solution in 70% ethanol. At the end of the staining procedure, seven worms cultured in microgravity and five worms cultured onboard the 1-G centrifuge were recovered and analyzed. Bright-field images were obtained using an Olympus BX51 microscope system with a DP73 digital camera (Olympus, Tokyo, Japan). The obtained images were converted to 8-bit gray-scale images and the extent of staining was quantitatively measured using ImageJ software version 1.49 (National Institutes of Health, Bethesda, MD, USA). The distribution of density across the nematode was converted to histograms using the “Histogram” function on the software.
6
Immunofluorescence Analysis of Angiogenic Markers
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The embedded brain tissues were sectioned to a thickness of 5 µm using a microtome (Leica, Nussloch, Germany). The sections were mounted on coated slides (#631-1349, Adhesion slides, Menzel Gläser, Polysine®, Thermo Fisher Scientific, UK), hydrated, and boiled in citrate buffer (pH 6.0) for antigen retrieval. After washing twice with 0.1% Triton X-100/PBS for 5 min, the sections were blocked with 1.5% normal horse serum for 1 h at RT to inhibit non-specific signals. The sections were subsequently incubated overnight at 4°C with anti-VEGFA (1:200, #SC1836, Santa Cruz Biotechnology), anti-CD31 (1:200, # AF3628, R&D Systems, MN, USA), anti-p-MAPK1 (#9102, Cell Signaling Technology, Inc), and anti-SP-1 antibodies, followed by incubation with the appropriate secondary antibodies for 1 h at RT. The slides were observed using an Olympus BX53 fluorescence microscope with DP73 digital camera (Olympus, Tokyo, Japan). Three fields were arbitrarily photographed for each tumor, and the average value of the three fields was used as a representative value for one tumor. The fluorescence intensity was analyzed by Image J using the following formula: Corrected Total Cell Fluorescence (CTCF) = Integrated Density − (Area of selected cell × Mean fluorescence of background readings).
7
Isolation and Characterization of AaDXS2 Promoter
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Genomic DNA of A. annua was isolated using the CTAB method. A genome walking method, that is, fusion primer and nested integrated PCR (Wang et al., 2011 (link)) was carried out to amplify the promoter of AaDXS2 (pAaDXS2). Primers used to amplify pAaDXS2 are listed in Supplementary Table S1 . The TSSP software was used to determine the transcription start site of AaDXS2 (Solovyev and Shahmuradov, 2003 (link)). The cis-elements in pAaDXS2 were analyzed using PlantCARE website1 and PLACE website2 .
To investigate the expression pattern of AaDXS2 in plants, the pAaDXS2 was cloned into pCAMBIA1391.Z to drive the expression of GUS gene. The pAaDXS2::GUS construct was introduced into Agrobacterium tumefaciens strain GV3101 and transformed into A. thaliana by the floral dip method (Zhang et al., 2006 (link)). Mature leaves and flowers of 45-day-old arabidopsis seedlings as well as siliques from 2-month-old transgenic A. thaliana were used for GUS histochemical staining as described previously (Jefferson et al., 1987 (link)). GUS stained tissues were observed under Olympus SZX16 microscope, and pictures were taken using Olympus DP73 digital camera.
To investigate the expression pattern of AaDXS2 in plants, the pAaDXS2 was cloned into pCAMBIA1391.Z to drive the expression of GUS gene. The pAaDXS2::GUS construct was introduced into Agrobacterium tumefaciens strain GV3101 and transformed into A. thaliana by the floral dip method (Zhang et al., 2006 (link)). Mature leaves and flowers of 45-day-old arabidopsis seedlings as well as siliques from 2-month-old transgenic A. thaliana were used for GUS histochemical staining as described previously (Jefferson et al., 1987 (link)). GUS stained tissues were observed under Olympus SZX16 microscope, and pictures were taken using Olympus DP73 digital camera.
8
Immunofluorescence Analysis of NF-κB p65 Translocation
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When cells were plated for the western blot experiments, coverslips were first added to the culture dishes, so that a portion of the treated cells could also be fixed and examined for p65 nuclear translocation. Prior to the addition of RIPA buffer, coverslips were removed from wells and fixed for 10 min with methanol/acetone (1:1 vol:vol) at -20° C. Cells were washed twice with 1X PBS, and immunofluorescence was carried out using primary antibodies against NFκB subunit p65 (1:200; cat. no. PA1-14298, Thermo Scientific) and secondary Alexa 488 antibody conjugates (1:200; Invitrogen, Grand Island, NY). Coverslips were mounted onto slides with Fluoromount G, and cells were imaged using an Olympus AX70 microscope equipped with an Olympus DP73 digital camera (Olympus, Center Valley, PA).
9
Immunofluorescence Characterization of Germ Cell and Oocyte Markers
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We used 4% paraformaldehyde/PBS at room temperature for 45 minutes to fix cells and permeabilization of cells was done by 0.2% Triton X-100 in PBS. The cells were blocked overnight with a blocking solution (5% milk and 0.05% Tween-20 in PBS) and then incubated with primary antibodies by using 1:100 dilution of monoclonal anti-DDX4 (ab13840, Abcam, USA), monoclonal anti-DAZL (ab34139, Abcam, USA), as a germ cell markers and monoclonal anti-SCP3 (ab15093, Abcam, Cambridge, UK), monoclonal anti-GDF9(ab93892, Abcam, USA) and monoclonal anti-GDF9B (ab108413, Abcam, Cambridge, UK) as oocyte like cells markers for 2.5 hrs. Afterward, cells were washed in PBS/Tween 20 (0.1%) (Sigma) three times and incubated with Alexa fluor 488 and 594 secondary antibodies (Sigma, USA) for 1hrs in a dark place at room temperature. Cells were again washed with PBS/Tween 20 (0.1%) three times. Finally, the cell nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich, D8417) and studied by Olympus DP73 digital camera associated with a fluorescence microscopy IX81 (U-MW-IB3). ImageJ software was used to assess the proportion of DDX4, DAZL, SCP3, GDF9, and GDF9B proteins in the resultant microphotographs.
10
Nrf2 Activation Assay in Cells
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Cells were seeded in dishes which had been pre-placed with glass covers and were treated with indicated doses of (+)-CLA for 18 h. Then, the glass covers were washed with PBS three times and fixed with −20 °C methanol/acetone (1:1) for 10 min. The glass covers were then incubated with anti-Nrf2 antibody at 4 °C overnight. After washing in PBS, the glass covers were incubated with Alexa Flour 594 and DAPI for the indicated time. An Olympus BX53 fluorescence microscope coupled to an Olympus DP73 digital camera (Tokyo, Japan) was employed to image the fluorescence signals.
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