RNA in situ hybridization was performed via RNAscope 2.5 HD Red Assay Kit (Advanced Cell Diagnostics, Cat: 322350) in accordance with manufacturer’s instructions. Fixed tissue sections were treated with the manufacturer’s Fresh Frozen Tissue Sample Preparation Protocol, fixed in chilled 4% PFA, dehydrated, and treated with H2O2 and Protease IV before probe hybridization. Paraffinized sections were deparaffinized and treated with H2O2 and Protease Plus prior to hybridization. Probes targeting SARS-CoV-2 spike (Cat: 848561), positive control Hs-PPIB (Cat: 313901), or negative control DapB (Cat: 310043) were hybridized followed by proprietary assay signal amplification and detection. Tissues were counterstained with Gill’s hematoxylin. An uninfected mouse was used as a negative control and stained in parallel. Tissues were visualized using an Olympus BX60 microscope and imaged with a Nikon (Model #) camera.
Bx60 microscope
Manufactured by Nikon
Sourced in United States
The BX60 is a high-quality microscope designed for laboratory use. It features a sturdy construction, precise optics, and a range of magnification capabilities to facilitate clear and detailed observation of samples. The BX60 is suitable for a variety of applications requiring accurate and reliable microscopic analysis.
Automatically generated - may contain errors
Lab products found in correlation
Smz1000 stereoscopic microscope, by Olympus (3 mentions)
Ds ri1 camera, by Nikon (3 mentions)
Rnascope 2.5 hd assay red kit, by Advanced Cell Diagnostics (2 mentions)
Ds ri1 camera head, by Nikon (2 mentions)
A1 confocal microscope, by Nikon (2 mentions)
S 3000n variable pressure scanning electron microscopy, by Hitachi (2 mentions)
Image pro plus software, by Nikon (1 mentions)
Dfc310 fx, by Leica (1 mentions)
Dab substrate chromogen, by Agilent Technologies (1 mentions)
Application suite, by Leica (1 mentions)
Envision dual link system hrp, by Agilent Technologies (1 mentions)
Dm1000 microscope, by Leica (1 mentions)
Nis elements ar, by Nikon (1 mentions)
Myeloperoxidase mpo, by Agilent Technologies (1 mentions)
Pcna clone pc10, by Agilent Technologies (1 mentions)
D700 digital camera, by Nikon (1 mentions)
S 3000n variable pressure scanning electron microscope, by Hitachi (1 mentions)
Ds ri1, by Nikon (1 mentions)
Pgem t easy vector, by Promega (1 mentions)
Digoxigenin labeling kit, by Roche (1 mentions)
12 protocols using bx60 microscope
1
SARS-CoV-2 Spike Protein in Situ Hybridization
2
Immunohistochemical Analysis of Aortic Root
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Cross-sections of the aortic root were stained with primary antibodies followed by HRP-conjugated secondary antibodies and developed with DAB substrate (brown). Images were captured under the Nikon Bx60 microscope connected to a Nikon DP70 camera with Cell-F imaging software (Soft Imaging System) and quantification was performed with Image Pro Plus Software.
3
Kidney Regeneration and Fibrogenesis Analysis
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In order assess kidney regeneration, paraffin-embedded sections were submitted to immunohistochemistry after heat mediated antigen retrieval and stained for Proliferating cell nuclear antigen (PCNA, clone PC10) (DAKO, USA) or Myeloperoxidase (MPO) (DAKO, USA), using a Horseradish Peroxidase (HRP)-conjugated secondary antibody EnVision™+ Dual Link System-HRP (Dako, EUA) revealed with 3,3’-diaminobenzidine (DAB)+ substrate-chromogen (Dako, EUA). In order to evaluate tissue fibrogenesis: Paraffin-embedded sections were submitted to picrosirius staining and subsequent polarization microscopy in order to distinguish collagen fibers. Kidney section images were captured at room temperature (21–25°C) at approximately 10 fields per kidney using an Olympus BX60 microscope and NIS-Elements F capture system (Nikon, Center Valley, PA, USA) or Leica DM 1000 microscope and Leica DFC310 FX (Leica, Wetzlar, Germany) capture system. Positive tissue staining was quantified through the use of Leica Application Suite (Leica, Wetzlar, Germany) or NIS-Elements AR (Nikon, Center Valley, PA, USA) software.
4
Multimodal Fluorescence Imaging of Cellular Structures
5
Quantifying Hippocampal Subregion Markers
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We chose the similar coronal sections from each group and captured images with an Olympus BX60 microscope equipped with a Nikon D700 digital camera. Using 20X objective, images of the CA1, CA3 and DG regions were captured. Image Pro Plus 6.0 image analysis software was used to analyse the images of 10 randomly selected sections from each group. Using a method described in the literature (15 (link)), the images of the hippocampal partition were processed. Before treatment, we corrected the space and optical density in microscale (the minimum scale of 0.01 mm) images and images of blank sections captured under the same conditions. The CA1 region was selected in a 200 × 100 μm2 area along the pyramidal layer, CA3 and DG regions were selected in 200 × 200 μm2 areas. The positive area and integrated optical density (IOD) of each slice was calculated, and calculate the MOD (MOD = IOD/area). The IOD represents the relative mRNA and protein levels. Data are presented as the means ± SD. P < 0.05 was considered to indicate statistically significant differences.
6
SARS-CoV-2 Spike Protein RNA In Situ Hybridization
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RNA in situ hybridization was performed via RNAscope 2.5 HD Red assay kit (Advanced Cell Diagnostics, Cat: 322350) in accordance with manufacturer’s instructions. Fixed tissue sections were treated with the manufacturer's Fresh Frozen Tissue Sample Preparation Protocol, fixed in chilled 4% PFA, dehydrated, and treated with H2O2 and Protease IV before probe hybridization. Paraffinized sections were deparaffinized and treated with H2O2 and Protease Plus prior to hybridization. Probes targeting SARS-CoV-2 spike (Cat: 848561), positive-control Hs-PPIB (Cat: 313901), or negative control DapB (Cat: 310043) were hybridized followed by proprietary assay signal amplification and detection. Tissues were counterstained with Gill’s hematoxylin. An uninfected mouse was used as a negative control and stained in parallel. Tissues were visualized using an Olympus BX60 microscope and imaged with a Nikon (Model number) camera.
7
Confocal and Electron Microscopy Imaging
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Confocal microscopy images were taken with a Nikon A1 confocal microscope. Samples were either live-imaged or fixed and sectioned as previously described [9 (link)]. Excitation and detection wavelengths for GFP, Venus, and DsRed were as previously described [9 (link), 32 (link)]. To detect FM4-64 and PI staining, a 514 nm laser line was used for excitation and a 561 nm long-pass filter was used for detection. The modified pseudo-Schiff-PI (mPS-PI) staining was performed as described and a 488 nm laser line was used for excitation and emission was collected at 520–720 nm [62 (link)]. DAPI staining was excited at 405 nm and detected in the 425–475 nm. Autofluorescence was excited at 488 nm or 514 nm and detected in the 660–700 nm range.
Optical photographs were taken with a Nikon SMZ1000 stereoscopic microscope or an Olympus BX60 microscope equipped with a Nikon DS-Ri1 camera. Scanning electron microscopy was performed using a Hitachi S-3000N variable pressure scanning electron microscope after standard tissue preparation [9 (link)].
Optical photographs were taken with a Nikon SMZ1000 stereoscopic microscope or an Olympus BX60 microscope equipped with a Nikon DS-Ri1 camera. Scanning electron microscopy was performed using a Hitachi S-3000N variable pressure scanning electron microscope after standard tissue preparation [9 (link)].
8
Leaf Morphology Phenotyping in Moss
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Gametophores (21 days old) with almost the same number of leaves were selected for phenotyping the leaf morphology. Leaves among the 10th (L10) to 14th (L14) leaf were dissected from each gametophore and photographed using an Olympus BX60 microscope equipped with a Nikon DS-Ri1 camera. Quantitative measurements were undertaken using the ImageJ software. The leaf length was measured as the distance between the leaf base and tip along the midline of the leaf. The leaf width was determined by the maximal leaf width across the widest part of the leaf. The leaf aspect ratio was calculated by dividing the leaf length by the leaf width for each leaf. Data analysis and visualization were performed in GraphPad Prism 9. At least 30 leaves from eight individuals were chosen for quantification.
9
GUS Activity Histochemical Detection
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The histochemical detection of GUS activity was performed according to a previously described method, with slight modifications (44 (link)). Briefly, the tissues were fixed in a solution containing 1% MES-KOH (pH 5.6), 0.3% formalin, and 0.3 M mannitol for 30 min at room temperature and washed three times with 50 mM NaH2PO4 (pH 7.0). Subsequently, the tissues were infiltrated for 30 min in 50 mM NaH2PO4 buffer (pH 7.0) supplemented with 0.5 mM 5-bromo-4-chloro-3-indolyl β-d -glucuronide (X-Gluc), 0.5 mM K3Fe(CN)6, 0.5 mM K4Fe(CN)6, and 0.05% Triton X-100 and then incubated at 37°C in the dark for 24 to 48 hours. The tissues were postfixed in 5% formalin and soaked in 5% acetic acid for 10 min at each stage. After undergoing a dehydration process in a graded ethanol series to 100%, the tissues were ready for further examination. Images were taken using an Olympus BX60 microscope equipped with a Nikon DS-Ri1 camera.
10
Characterizing SlWOX1 Expression via In Situ Hybridization
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A 773-bp coding sequence of SlWOX1 was PCR-amplified from M82 cDNA and cloned into pGEM-T Easy vectors (Promega, Cat. no. A1360) for in vitro transcription using a digoxigenin labeling kit (Roche, Cat. no. 11277073910) . Anti-sense and sense probes were synthetized using SP6 (Promega, Cat. no. P1085) and T7 RNA polymerase (Promega, Cat. no. P2075), respectively. The long probes were then hydrolyzed to an average length of 150 bp and resuspended in 50% formamide at the desired concentrations. Primers for amplifying probes are listed in Table S1.
In situ hybridization experiments were performed as previously described (Zhang et al., 2017) . The paraffin sections were 8 lm thick. Both the hybridization and washing were performed at 55°C, and staining was conducted at room temperature. Optical photographs of in situ hybridization sections were taken with an Olympus BX60 microscope equipped with a Nikon DS-Ri1 camera.
In situ hybridization experiments were performed as previously described (Zhang et al., 2017) . The paraffin sections were 8 lm thick. Both the hybridization and washing were performed at 55°C, and staining was conducted at room temperature. Optical photographs of in situ hybridization sections were taken with an Olympus BX60 microscope equipped with a Nikon DS-Ri1 camera.
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