Two additional teeth from each group were prepared for confocal laser scanning microscopy (CLSM) to observe the interaction between the adhesive system and internal dentinal fluids at the dentin-resin layer. Three drops of each adhesive system were mixed with one grain of tetramethylrhodamine B isothiocyanate (Sigma-Aldrich) to form a homogeneous solution. Tooth samples were restored using the methodology described above for each of the groups. After 24 h of storage in water, the pulpal chambers were filled with a solution of 0.1% fluorescein sodium (Sigma-Aldrich) for 4 h. The restored tooth segments were cut perpendicular to the adhesive interface using a slow-speed precision saw to obtain 2–3 slabs with 1.0 mm thickness from the center of the tooth. The samples were evaluated under a CLSM (LSM 5 Pascal, Zeiss, Jena, Germany) and images of the adhesive interface with 63 times of magnification at 1024 × 1024 pixels and 2.0 μs/pixel speed were obtained.
Tetramethyl rhodamine b isothiocyanate
Manufactured by Merck Group
Tetramethyl rhodamine B isothiocyanate is a fluorescent dye used in various laboratory applications. It is a derivative of rhodamine B, a common fluorescent tracer. The core function of this dye is to provide a fluorescent label for biomolecules, such as proteins or nucleic acids, enabling their detection and visualization in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Sodium fluorescein, by Merck Group (1 mentions)
Lsm 5 pascal, by Zeiss (1 mentions)
Endothelial growth medium, by Thermo Fisher Scientific (1 mentions)
Matrigel matrix, by Merck Group (1 mentions)
Polymyxin b agarose, by Merck Group (1 mentions)
Mcp 1, by Merck Group (1 mentions)
Transwell tissue culture insert, by Corning (1 mentions)
24 well plate, by Corning (1 mentions)
Calcein am, by Merck Group (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
5 protocols using tetramethyl rhodamine b isothiocyanate
1
Visualizing Dentin-Resin Interface Using CLSM
2
Transwell-Based Endothelial Cell Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transwell tissue culture inserts (diameter, 6.5 mm; pore size, 5.0 µm) and 24-well plates were purchased from Corning Costar Corp (Corning, NY). RPMI 1640 medium and endothelial growth medium were obtained from Gibco (Grand Island, NY). Heat-inactivated fetal bovine serum was obtained from HyClone (Logan, Utah). Mouse anti-human claudin-1 was obtained from Cell Signaling Technology (America, Boston). The Limulus amebocyte lysate test kit, polymyxin B-agarose, anti-human ICAM-1 monoclonal antibody (MAb), anti-human E-selectin (MAb), Calcein AM, phalloidin, tetramethylrhodamine bisothiocyanate, matrigel matrix, and MCP-1 were purchased from Sigma-Aldrich (America, St Louis, MO). Recombinant T. pallidum protein Tp0965(rTp0965)was purified on nickel-nitrilotriacetic acid (Ni-NTA) chromatographic column from E. coli lysates frozen in our laboratory. The human MCP-1 enzyme-linked immunosorbent assay (ELISA) kit was purchased from R&D Systems, Inc. (Minneapolis, MN). Anti-mouse immunoglobulin G and horseradish peroxidase (HRP)-tagged antibody were purchased from Amersham (Piscataway, NJ). Anti-rabbit HRP-tagged antibody was purchased from Zymed (San Francisco, CA).
3
Phalloidin Staining of Zebrafish Embryos
4
Actin Cytoskeleton Visualization of eq-ASCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The morphology of the eq-ASCs on the scaffolds
was evaluated with phalloidin staining of the actin
cytoskeleton. Briefly, cells were fixed in formaldehyde 4%,
permeabilized using triton 0.2%, and sequentially stained
with the phalloidin-TRITC label (Tetramethylrhodamine
B isothiocyanate; Sigma) and 4ˊ, 6-diamidino-2-
phenylindole (DAPI, Sigma). Finally, fluorescence
images were obtained using fluorescence microscopy
(Olympus, BX51, Japan).
was evaluated with phalloidin staining of the actin
cytoskeleton. Briefly, cells were fixed in formaldehyde 4%,
permeabilized using triton 0.2%, and sequentially stained
with the phalloidin-TRITC label (Tetramethylrhodamine
B isothiocyanate; Sigma) and 4ˊ, 6-diamidino-2-
phenylindole (DAPI, Sigma). Finally, fluorescence
images were obtained using fluorescence microscopy
(Olympus, BX51, Japan).
5
Phalloidin Staining of Zebrafish Embryos
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phalloidin stainings were performed according to previous reports.30 31 (link)
shroom4 MO-injected zfl were fixed at 5 dpf in 4% PFA overnight at 4°C and subsequently washed three times for 5 min in phosphate-buffered saline (PBS) with 0.1% Tween-20 (PBST) and 5 min in PBS-Tx (2% Triton X-100). Afterwards, zfl were incubated for 2 hours in PBS-Tx to allow permeabilisation. Stainings with 2 µg/mL tetramethyl rhodamine B isothiocyanate (Sigma Aldrich, P1951) were conducted overnight at 4°C. Stained zfl were briefly washed in PBS before applying DAPI (ACDBio, RNAscope DAPI, 320858) overnight at 4°C. Zfl were washed in PBS before imaging.
shroom4 MO-injected zfl were fixed at 5 dpf in 4% PFA overnight at 4°C and subsequently washed three times for 5 min in phosphate-buffered saline (PBS) with 0.1% Tween-20 (PBST) and 5 min in PBS-Tx (2% Triton X-100). Afterwards, zfl were incubated for 2 hours in PBS-Tx to allow permeabilisation. Stainings with 2 µg/mL tetramethyl rhodamine B isothiocyanate (Sigma Aldrich, P1951) were conducted overnight at 4°C. Stained zfl were briefly washed in PBS before applying DAPI (ACDBio, RNAscope DAPI, 320858) overnight at 4°C. Zfl were washed in PBS before imaging.
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!