| ID |
3 |
| date of publication |
2025/3/28
LM-GlycomeAtlas
|
| Species |
Mouse |
| Supplier |
Leica |
| Instrument |
LMD7000 |
| Observation Type |
Bright-field |
| Detail Description |
Tissue dissection were performed using an LMD system (LMD7000, Leica Microsystems) attached to a microscope (DM6000B, Leica Microsystems). Tissue fragments (total area: 0.5 mm^2) from hematoxylin-stained sections were collected into a tube. |
| Lectin Array Chip |
download (Excel) |
| Sample preparation |
20 µL of protein extracts were fluorescently labeled with Cy3-succinimidyl ester (Cy3-SE; 10 µg of protein equivalent; GE Healthcare, Buckingham- shire, UK) for 1 h at RT in the dark. After adjusting the volume to 100 µL with probing buffer (25 mM Tris-HCl at pH 7.5, containing 137 mM NaCl, 2.7 mM KCl, 500 mM glycine, 1 mM CaCl2, 1 mM MnCl2, and 1% Triton X-100), the sample solutions were incubated for 2 h at RT to quench the excess reagent. |
| Supplier of instrument |
GlycoTechnica |
| Instrument |
GlycoStation Reader1200 |
| Data acquisition |
After appropriate dilution with the probing buffer, the sample solutions (0.075-0.3 mm^2 equivalent) were added to wells (60 µL/well) on the lectin array chip (LecChip; GlycoTechnica, Yokohama, Japan) containing triplicate spots of 45 lectins. After incubating overnight at 20 °C and washing three times with probing buffer, the array chip was scanned using an evanescent-field fluorescence scanner (GlycoStation Reader 1200; GlycoTechnica). All data were analyzed using GlycoStation Tools Pro Suite 1.5 software (GlycoTechnica). |
| Image analysis software |
After appropriate dilution with the probing buffer, the sample solutions (0.075–0.3 mm2 equivalent) were added to wells (60 µL/well) on the lectin array chip (LecChip; GlycoTechnica, Yokohama, Japan) containing triplicate spots of 45 lectins. After incubating overnight at 20 °C and washing three times with probing buffer, the array chip was scanned using an evanescent-field fluorescence scanner (GlycoStation Reader 1200; GlycoTechnica). All data were analyzed using GlycoStation Tools Pro Suite 1.5 software (GlycoTechnica). |
| Data processing and statistical analysis |
The net intensity was calculated by subtracting the background from the signal intensity. The data under appropriate gain conditions providing net intensities below 40,000 for all spots were used to obtain the glycomic pro- files, which were presented as the relative signal intensities of the 45 lectins normalized to the mean value of all the lection signals. |
