A previously described intravital microscopy method was used [10 (link), 81 (link)]. Female 7-week old C57BL/6 mice were anesthetized with a ketamine/xylazine. Their peritoneal cavity was opened via midline abdominal incision, and their mesenteric circulation was exposed. To activate mesenteric endothelium, 5 μl of a 10 mM Ca2+-ionophore A23187 was topically applied to the surface of the mesentery. Afterwards, GFP-expressing bacteria (1.2×108 CFU/mouse in 50 μl PBS) were injected intravenously through the retro-orbital plexus. After 30 minutes, all visible adhesion of S. aureus to mesenteric vasculature was assessed and captured with a Nikon upright microscope with a digital camera. If no adhesion was observed, representative images of vessels were captured.
Upright microscope
Manufactured by Nikon
Sourced in Japan, United States
The Nikon Upright Microscope is a versatile laboratory instrument designed for a wide range of microscopic observations. It features a stable, upright stand that provides a reliable platform for precise imaging and analysis. The microscope incorporates high-quality optics, allowing users to examine samples with clarity and precision. Its core function is to magnify and observe small-scale specimens, enabling detailed study and examination.
Automatically generated - may contain errors
Lab products found in correlation
Alexa 488 labeled phalloidin, by Thermo Fisher Scientific (3 mentions)
Triton x 100, by Merck Group (3 mentions)
Hoechst 33342, by Merck Group (3 mentions)
Bovine serum albumin (bsa), by Merck Group (3 mentions)
One step hrp polymer, by Abcam (2 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (2 mentions)
Plan fluor 20, by Nikon (2 mentions)
S plan fluor 40, by Nikon (2 mentions)
Zli 9557, by ZSGB-BIO (1 mentions)
Ds ri2 color ccd, by Nikon (1 mentions)
Ff875 di01, by IDEX Corporation (1 mentions)
Et900lp, by Chroma Technology (1 mentions)
Chemray 240 automatic biochemical analyzer, by Rayto (1 mentions)
Matrigel, by BD (1 mentions)
Imagej, by Nikon (1 mentions)
Transwell chamber, by Merck Group (1 mentions)
Matrigel, by Corning (1 mentions)
Crystal violet, by Beyotime (1 mentions)
Image pro plus software, by Media Cybernetics (1 mentions)
Mitotracker, by Thermo Fisher Scientific (1 mentions)
44 protocols using upright microscope
1
Intravital Imaging of Bacterial Adhesion in Mouse Mesentery
2
Immunohistochemical Detection of LC3 and F4/80 in Lung and Tumor Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemistry for detection of the LC3 protein and the macrophage marker F4/80 was carried out using standard protocols. Briefly, unstained sections of lungs or tumors were deparaffinized and rehydrated, and then incubated in the peroxidase blocking reagent (BioVision cat #K405‐50). Antigen retrieval was performed by boiling the sections in sodium citrate for 20 min. To decrease background staining, the slides were incubated for 1 h in the mouse on mouse blocking reagent (Vector Labs MKB‐2213‐1), followed by overnight incubation with the primary antibodies (LC3B Cell Signaling 3868S rabbit polyclonal, 1:300; or F4/80 Cell Signaling 70076S rabbit monoclonal, 1:200). Next day, the slides were washed and incubated with One‐Step HRP polymer (BioVision cat #K405‐50) for 30 min at RT. The slides were then washed several times and then incubated with the DAB chromogen for 10 min at RT, followed by several washing steps and quick counterstain with Hematoxylin. Slides were then mounted and visualized under the upright microscope (Nikon).
3
Colon Histopathological Scoring Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The colon was fixed in 4% paraformaldehyde, dehydrated, and transparentized. After paraffin embedding, the colon was selected, dewaxed and stained with HE, transparentized and sealed, and observed under an upright microscope (Nikon Japan), and the histopathological scoring standard was in accordance with Murano standard: degree of inflammatory infiltration (0-5), degree of crypt injury (0-4), degree of ulcer (0-3), and whether there is edema (0 or 1) .
4
TUNEL Staining of HUASMCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was performed according to the manufacturer’s instructions (Roche #12156792910). In short, HUASMCs were plated in dishes (60 mm) with slides, and after AngII and/or ANGPT8 siRNA treatment, cells were embedded in paraffin for 30 min and then treated with 0.01% Triton for 2 min on ice. Slides were then treated with TUNEL reaction mixture in a humidified chamber and washed with PBS. Finally, slides were mounted using Vectashield mounting medium with DAPI (ZLI-9557, ZSGB-BIO, Beijing, China). Images were obtained with a Ni-UNikon Upright Microscope equipped with a DS-Ri2 color CCD (Nikon, Tokyo, Japan).
5
Selective Excitation of (6,5) SWCNTs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Nanotubes were excited by a tunable Ti:Sa laser to preferentially excite (6,5) SWCNTs at the resonance of the dark K-momentum exciton. The beam was focused into the back aperture of a high NA objective (60×, NA 1.0) mounted on an upright microscope (Nikon, Tokyo, Japan), with an excitation intensity of 10 kW/cm2 of circularly polarized light at the sample. The fluorescence was collected with the same objective and imaged on a low noise EMCCD camera (Roper Scientific SAS, Evry, France) to produce wide-field images of individual SWCNTs. A dichroic mirror (FF875-Di01, Semrock, Rochester, NY, USA) and the combination of long- and short-pass emission filters (ET900LP, Chroma Technology Corp., Bellows Falls, VT, USA; FESH1000, Thorlabs SAS, Maisons-Laffitte, France) were used in order to illuminate and detect the (6,5) SWCNTs’ emitted fluorescence. Images of SWCNTs were recorded with 30 ms integration time per frame.
6
Liver Histology and Biomarkers in Rats
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The isolated rat livers were fixed with 4% paraformaldehyde, embedded in paraffin, and sliced. Slices were stained with hematoxylin and eosin (H&E), and masson-stained. Histological changes were evaluated by randomly selecting visual fields using an upright microscope (Nikon, Japan). The levels of alanine transaminase (ALT), aspartate transaminase (AST), gamma-glutamyl transferase (γ-GGT), total bilirubin (TBIL) and direct bilirubin (DBIL) were measured using a Chemray 240 Automatic Biochemical Analyzer (Rayto Life and Analytical Sciences Co., Ltd., Shenzhen, China). Albumin (ALB) was evaluated using commercial kits, according to the manufacturer’s instructions.
7
Cell Migration and Invasion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For migration assays, 5 × 104 cells in 200 μL of serum-free media were seeded in the upper chamber of an insert (8 -μm pore size, Corning, USA). For invasion assays, 2 × 105 cells in 200 μL of serum-free media were seeded into the upper chamber of an insert coated with Matrigel (BD Bioscience, USA). Five-hundred microliters of media containing 10% FBS were added to the lower chamber. After 24 h of incubation, the cells remaining on the top surface of the membrane were gently removed with a cotton swab, while the cells migrating or invading through the membrane were fixed with methanol and stained with 0.1% crystal violet. The entire membrane surface was imaged, and the cells were counted using an upright microscope (Nikon, Japan). Each experiment was performed in triplicate.
8
Cresyl Violet Staining of Brain Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen sections of brain tissue were fixed with 4% paraformaldehyde for 15 min and washed three times with PBS. Cresyl Violet Staining (Solarbio, China) was added to brain slices and incubated at 55°C for 1 h. Wash off the excess staining and differentiate by Nissl differentiation solution for a few seconds until the background is nearly colorless. Seal with glycerin jelly mounting medium and photographed with upright microscope (Nikon, Japan) and analyzed with ImageJ.
9
Histopathological Analysis of Vital Organs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mice were euthanized on 42 days after first immunization for tissue processing. Vital organs such as lungs, hearts, livers, kidneys, and spleen were harvested and fixed in 10% phosphate‐buffered formalin for 7 days, embedded in paraffin and sectioned at 3 μm thickness. Sequential sections were stained with hematoxylin and eosin to assess pathology and organ damage. Stained slides were scanned with an upright microscope (Nikon).
10
Cell Migration and Invasion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell migration and invasion were assessed using Transwell chambers (8.0 μm pore size; EMD Millipore, Billerica, MA, USA) with or without Matrigel (diluted 1:9) (Corning Inc., USA), respectively. For the migration assay, infected Hep-3B cells (4×105) or Huh-7 cells (2×105) were resuspended in 200 μL of serum-free MEM (Minimum Essential Medium) and seeded into the upper chamber, while 500 μL of DMEM containing 10% serum was added to the lower chamber. For the invasion assay, cell-containing medium was seeded into the Matrigel-coated upper chamber, and 500 μL of DMEM containing 10% serum was added to the lower chamber. After 24 h, the cells in the upper chamber were removed with cotton swabs, while those on the lower membrane were fixed in 4% paraformaldehyde for 30 min and then stained with 0.1% crystal violet (Beyotime, Jiangsu, China) for 30 min. Cells were counted under an upright microscope (Nikon, Tokyo, Japan, 200× magnification).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!