The fecal bacteria solution was stained with 10 µg/ml CM-DiI fluorescent dye (Thermo Fisher Scientific, Waltham, MA, USA) for 30 min at 37 °C. Mice were gavaged with 200 μl stained fecal bacteria solution. Fresh feces of recipient mice were collected on seven days after intragastric administration. The feces were examined by confocal microscope (Nikon MODEL ECLIPSE Ni-E).
Cm dii fluorescent dye
Manufactured by Thermo Fisher Scientific
Sourced in United States
CM-DiI is a fluorescent dye used for labeling and tracking cells. It is lipophilic in nature and can be incorporated into the cell membrane. The dye emits fluorescence that can be detected using appropriate optical filters.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using cm dii fluorescent dye
1
Tracking Gut Bacteria Colonization
2
KNGT Fecal Microbiota Transfer in db/db Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen fecal bacteria solutions were thawed for 2–4 h in an ice bath prior to FMT. The fecal bacteria solution was then stained with 10 µg/mL CM-DiI fluorescent dye (Thermo Fisher Scientific, Waltham, MA, USA) for 30 min at 37 °C. Each db/db + KNGT mouse was gavaged with 0.2 mL of the stained fecal bacteria solution [25 (link)]. Mice gavaged with the same amount of PBS were used as controls. Fresh feces of treated and control mice were collected in sterile tubes on days 3, 7, and 14, respectively, after intragastric administration. The feces were examined by using an Olympus IX73 microscope (Olympus, Tokyo, Japan). Detection of fluorescent signals in the fecal bacteria solution of db/db + KNGT mice on day 14 confirmed successful colonization of KNGT fecal bacteria in the db/db mice.
3
Tracking BM-MSCs and WJ-MSCs in Rats
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The BM-MSCs or WJ-MSCs were labeled using CM-DiI fluorescent dye (Invitrogen) in order to track them in vivo in rats. Briefly, the cells were resuspended in KO-DMEM (Gibco) along with 5 μM CM-DiI and incubated for 30 min at 37 °C. The excess dye was removed by washing with DPBS, and the cells were resuspended in Plasmalyte A and injected at a dose of 10 × 106 cells/rat through the tail vein. Animals received vehicle alone to serve as normal controls. The DiI-labeled cells were confirmed ex vivo in liver tissues by fluorescence microscopy after 30 days of cell administration.
4
Isolation and Characterization of Exosomes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After 72h of HO-1/BMMSC culture in exosome-free serum medium, debris and dead cells in the medium were removed by centrifugation at 250×g for 15min and 3000×g for 30min and then filtered through a 0.22μm filter. The medium was then subjected to ultracentrifugation at 110,000×g for 140min at 4°C. Exosomes were resuspended in sterile phosphate-buffered saline (PBS).
Exosome morphology was observed using a transmission electron microscope (Hitachi-HT7700, Tokyo, Japan). The exosomal particle size was determined using nanoparticle tracking analysis (Malvern-NS300, Malvern, UK). The levels of exosome-specific proteins CD9, CD63, tumor susceptibility 101 (TSG101), and calnexin (negative control; ProteinTech, Wuhan, China) were detected using western blotting.
To monitor exosome trafficking, exosomes were labeled with chloromethyl-1,1′-dioctadecyl-3,3,3′3′-tetramethylindocarbocyanine perchlorate (CM-DiI fluorescent dye, Invitrogen, Waltham, MA, USA) for 15 min in the dark at 4°C. After CM-DiI staining, the exosomes were washed in PBS and collected by ultracentrifugation (110,000×g for 70min) at 4°C. Finally, CM-DiI-labeled exosomes were resuspended in PBS. The rat liver cell line IAR20 was cocultured with the labeled exosomes for 6 h, and the uptake of exosomes was observed using a confocal microscope (FluoView™-FV1000, Olympus, Tokyo, Japan).
Exosome morphology was observed using a transmission electron microscope (Hitachi-HT7700, Tokyo, Japan). The exosomal particle size was determined using nanoparticle tracking analysis (Malvern-NS300, Malvern, UK). The levels of exosome-specific proteins CD9, CD63, tumor susceptibility 101 (TSG101), and calnexin (negative control; ProteinTech, Wuhan, China) were detected using western blotting.
To monitor exosome trafficking, exosomes were labeled with chloromethyl-1,1′-dioctadecyl-3,3,3′3′-tetramethylindocarbocyanine perchlorate (CM-DiI fluorescent dye, Invitrogen, Waltham, MA, USA) for 15 min in the dark at 4°C. After CM-DiI staining, the exosomes were washed in PBS and collected by ultracentrifugation (110,000×g for 70min) at 4°C. Finally, CM-DiI-labeled exosomes were resuspended in PBS. The rat liver cell line IAR20 was cocultured with the labeled exosomes for 6 h, and the uptake of exosomes was observed using a confocal microscope (FluoView™-FV1000, Olympus, Tokyo, Japan).
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!