Soft palate tissues were harvested from OSA and control groups, then fixed with paraformaldehyde and embedded in paraffin. For histology, paraffin sections (4 μm thick) were stained with hematoxylin and eosin. For immunohistochemistry, paraffin sections were de‐paraffinized, rehydrated and antigen retrieved with incubation in 0.01 m citrate buffer (pH 6.0) at 121 °C per 100 kpa for 2 min. Slides were incubated with primary antibodies including anti‐CD68 (dilution 1 : 200; ab125212; Abcam, Cambridge, UK), anti‐HMGB1 (dilution 1 : 200; ab18256; Abcam), anti‐TLR4 (dilution 1 : 200; GB11519; Servicebio, Woburn, MA, USA) and anti‐CD31 (dilution 1 : 200; ab28364; Abcam) at 4 °C overnight. Then secondary antibody conjugated with fluorescent dye were incubated at 37 °C for 30 min. Nuclei were stained with 4′,6‐diamidino‐2‐phenylindole (DAPI) (Sigma‐Aldrich, St Louis, MO, USA). Images were acquired with a 50i Nikon fluorescence microscope (Nikon, Tokyo, Japan) and processed with photoshop cs 4 software (Adobe Sytems, San Jose, CA, USA).
50i fluorescence microscope
Manufactured by Nikon
Sourced in United States
The Nikon 50i fluorescence microscope is a versatile instrument designed for high-performance fluorescence imaging. It features a powerful illumination system, advanced optics, and a user-friendly interface. The 50i is capable of capturing detailed, high-quality fluorescence images for a wide range of applications.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Merck Group (4 mentions)
Ab28364, by Abcam (2 mentions)
Ab18256, by Abcam (1 mentions)
Ab125212, by Abcam (1 mentions)
Gb11519, by Wuhan Servicebio Technology (1 mentions)
Bromodeoxyuridine (brdu), by Merck Group (1 mentions)
Fluorescently tagged secondary antibodies, by Cell Signaling Technology (1 mentions)
Hematoxylin, by Merck Group (1 mentions)
Hoechst 33258, by Merck Group (1 mentions)
Vectastain dab kit, by Vector Laboratories (1 mentions)
Ab15580, by Abcam (1 mentions)
Ab134175, by Abcam (1 mentions)
Ab181591, by Abcam (1 mentions)
Ab81299, by Abcam (1 mentions)
Ab52189, by Abcam (1 mentions)
Ab6326, by Abcam (1 mentions)
Neutral balsam, by Solarbio (1 mentions)
Ab2910, by Abcam (1 mentions)
Ab14196, by Abcam (1 mentions)
6 protocols using 50i fluorescence microscope
1
Histological and Immunohistochemical Analysis of Soft Palate Tissues in OSA
2
Hypoxia effects on iHepSC proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The effect of hypoxia on the proliferative activity of iHepSCs was investigated by bromodeoxyuridine (BrdU, Sigma, St. Louis, MO, USA) incorporation. After 24 h incubation under the hypoxic condition, iHepSCs were labeled with 10 M BrdU for 2 h, fixed in 4% paraformaldehyde for 15 min, washed with PBS for 5 min × 3, and incubated in 2 N hydrochloric acid (HCl) for 30 min at 37°C and in 0.1 M sodium borate (pH 8.5) for 10 min (exclusively in BrdU incorporation assay). Cells were washed with PBS-Tween, blocked with 1% bovine serum albumin (BSA) for 30 min at room temperature, and incubated overnight at 4°C with primary antibodies in PBS containing 0.1% Triton X-100 and 1% BSA. After washing in PBS, cells were reacted with the fluorescent-labeled secondary antibody for 1 h at 37°C. The nucleus was counterstained with Hoechst 33342. Images were obtained with a 50i Nikon fluorescence microscope (Nikon). The information about the antibodies is listed in Supplemental Table
2 .
3
Immunofluorescent Staining of FAIM2 in H5V Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H5V cells were fixed for 20 minutes using PBS containing 4% paraformaldehyde (Guangzhou, China), and then blocked for 30 minutes using blocking buffer (RiboBio). Cells were incubated with primary antibody rat anti-mouse FAIM2 (Abcam) at 4°C overnight, then incubated with fluorescently-tagged secondary antibodies (Cell Signaling Technology) for 30 minutes at 37°C. Nuclei were stained using 4′, 6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich, St. Louis, MO, USA). Images were captured by a 50i Nikon fluorescence microscope (Nikon, Melville, NY, USA), and processed with Adobe Photoshop CS4 software (San Jose, CA, USA). The number of Fas, FADD, or Caspase3 positive cells was determined by the number of DAPI-stained positive cells.
4
Immunohistochemical and Immunocytochemical Analysis of Cell Cycle Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemical staining for H2A histone family, member X (γ-H2AX, Abcam, ab81299), BrdU (Abcam, ab6326), Ki67 (Abcam, ab15580), Cyclin A2 (Abcam, ab181591), Cyclin E1 (Abcam, ab52189) and Cyclin D1 (Abcam, ab134175) were performed on 4% paraformaldehyde-fixed liver sections. 5μm-thick slices were incubated with primary antibodies at 4? overnight followed by biotinylated secondary antibody and the avidin/biotin horseradish peroxidase system (Vectastain DAB Kit; Vector Laboratories, Burlingame, CA). The nuclei were counterstained with hematoxylin (Sigma-Aldrich, St. Louis, MO) and the sections were covered in neutral balsam (Solarbio, Beijing, CHN). Immuno-fluorescent staining for β-catenin (Sigma-Aldrich, MAB2081) and PHH3 (Roche, 760-4591) were detected on paraffin-embedded liver section. Slides were incubated with primary antibodies at 4 °C overnight and then with secondary antibodies conjugated with fluorescent dye at 37°C for 30 min.
Immunocytochemical staining for BrdU (Abcam, ab6326) and Ki67(Abcam, ab15580), cells were fixed, permeabilized and blocked, then incubated with primary antibody, followed by fluorescence-tagged secondary antibodies. Nuclei were counterstained with Hoechst 33258 (Sigma-Aldrich, St. Louis, MO). Images were acquired with a 50i Nikon fluorescence microscope (Nikon, Melville, NY).
Immunocytochemical staining for BrdU (Abcam, ab6326) and Ki67(Abcam, ab15580), cells were fixed, permeabilized and blocked, then incubated with primary antibody, followed by fluorescence-tagged secondary antibodies. Nuclei were counterstained with Hoechst 33258 (Sigma-Aldrich, St. Louis, MO). Images were acquired with a 50i Nikon fluorescence microscope (Nikon, Melville, NY).
5
Histological Analysis of Rat Soft Palate
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Soft palate harvested from rats in both groups were fixed with paraformaldehyde, and later embedded in paraffin. Paraffin sections (4 μm thick) were then stained with hematoxylin and eosin (H & E) to examine the histological change of mucosa and muscles in soft palate. For immunohistochemistry, paraffin sections were de-paraffinized, rehydrated with xylene and ethanol, and antigen was retrieved by incubation in 0.01 M citrate buffer (pH 6.0) at 121°C/100 kpa in a pressure cooker for 3 minutes. Slides were incubated overnight with primary antibodies including anti-annexin V (1: 200, ab14196 Abcam), anti-CD31 (1: 200, ab28364 Abcam), anti-caveolin-1 (1: 100, ab2910 Abcam), and anti-LC3B (1: 200, 3868 R&D) at 4°C, then incubated with secondary antibody conjugated with fluorescent dye at 37°C for 30 minutes. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich, St. Louis, MO, USA). Images were acquired with a 50i Nikon fluorescence microscope (Nikon, Melville, NY, USA) and analyzed using Adobe Photoshop CS4 software (San Jose, CA, USA).
6
Immunocytochemistry of Tight Junction Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We used 4% formaldehyde to fix MLVECs for 15 min and permeabilized by 0.1% Triton X-100 for 15 min. We incubated fixed cells in 10% BSA for 1 h, and cells were then reacted with ZO-1 primary antibodies at a dilution of 1: 50 at 37°C for 1.5 h. After washing to remove nonspecific binding, cells were incubated with anti-rabbit IgG-FITC for ZO-1 used in 1: 1000 dilution at 37°C under dark conditions for 1 h. Cells were subjected to 3 PBS washes and exposed to 4′6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich) to stain the nuclei. We captured the ICC images using a 50i Nikon fluorescence microscope (Nikon, Melville, NY).
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!