|
| Status |
Public on Mar 07, 2023 |
| Title |
Control=-1-10-22 | 003 | CD45 |
| Sample type |
protein |
| |
|
| Source name |
gastrocnemius muscle
|
| Organism |
Mus musculus |
| Characteristics |
strain: CD-1 strain
treatment: 50 ul PBS
roi name: High Desmin (HD)
segmentation (aoi): CD45
merge: Control-3-HD
group: Control
|
| Treatment protocol |
Groups of mice (CD-1 strain, 18-20 grams) received an intramuscular injection, in the right gastrocnemius, of 30 µg venom. Control mice were injected with 50 µL PBS only. At the time intervals of 1 h, 6 h, 24 h following injection, groups of three mice were sacrificed by CO2 inhalation, and the tissue was excised and added to 10% formalin solution in water.
|
| Growth protocol |
Groups of mice (CD-1 strain, 18-20 grams)
|
| Extracted molecule |
protein |
| Extraction protocol |
Muscle tissue sections (4 µm thick) were deparaffinized, subjected to antigen retrieval, and incubated overnight with three fluorescent-labeled visualization antibodies to detect muscle fibers (desmin), leukocytes (CD45), and nuclear stain (Syto 83), along with a cocktail of 48 oligonucleotide-labeled antibodies to detect a variety of cellular markers
|
| Label |
Barcode probe
|
| Label protocol |
Eighteen regions of interest (ROI) from 416 to 652 μm diameter were selected based on desmin staining intensity to generate Low Desmin (LD) and High Desmin (HD) areas
|
| |
|
| Hybridization protocol |
Once the 18 ROIs were processed, indexing oligos were hybridized to NanoString optical barcodes for digital counting on the nCounter
|
| Scan protocol |
UV light illumination of the ROIs, the eluent was collected via microcapillary aspiration and transferred into individual wells of a microtiter plate.
|
| Description |
Control-3-HD - Sum
Protein Count
three fluorescent-labeled visualization antibodies to detect muscle fibers (desmin), leu-kocytes (CD45), and nuclear stain (Syto 83), along with a cocktail of 48 oligonucleo-tide-labeled antibodies to detect a variety of cellular markers (Supplementary Table S1). Eighteen regions of interest (ROI) from 416 to 652 μm diameter were selected based on desmin staining intensity to generate Low Desmin (LD) and High Desmin (HD) areas
|
| Data processing |
Raw data coount was subjeted to Quality Conrtol (QC) analysis, data normalization, background exclusion, and imputation data
|
| |
|
| Submission date |
Jan 16, 2023 |
| Last update date |
Mar 07, 2023 |
| Contact name |
Ana Karina de Olliiveirra |
| E-mail(s) |
ankaoliv@gmail.com
|
| Phone |
434-924-1753
|
| Organization name |
UVA
|
| Department |
Department of Pathology
|
| Lab |
Spatial Biology Core
|
| Street address |
1340 Jefferson Park Avenue
|
| City |
Charlottesville |
| State/province |
VA |
| ZIP/Postal code |
22908 |
| Country |
USA |
| |
|
| Platform ID |
GPL33025 |
| Series (1) |
| GSE222977 |
Mapping the immune cell microenvironment by digital spatial profiling in muscle tissue injected with the venom of Daboia russelii |
|
