Cellular and Tissue Phenotyping Platform
The Cellular and Tissue Phenotyping Platform provides the entire scientific community with high-tech instrumentation.
Presentation
It covers flow cytometry, cell sorting, protein screening and sample preparation for cytometric analysis or sorting, mRNA sequencing (whole tissue RNAseq, single cell RNAseq, etc.) or single cell sequencing (single cell RNAseq).
Expertise
The Cellular and Tissue Phenotyping Platform of the Institute of Vision specializes in the following areas of expertise:
- Cell viability
- Apoptosis
- Caspase activity
- Mitochondrial membrane potential
- Cell proliferation
- Cell cycle
- Cytokines
- Antigen quantification
- Direct and indirect immunolabeling membrane or intracytoplasmic
- Cell characterization and phenotyping
- Bioinformatics analysis of sequencing data
Equipment
Flow Cytometer - Celesta SORP - Becton Dickinson - 4 lasers - 18 colors
The Celesta Sorp is equipped with the HTS (Hight Throughput Sampler) system, enabling fully automated high-throughput sample acquisition.
LASER | Filtres | Mirrors | Fluorochromes |
|---|---|---|---|
Violet (405 nm) | 450/40 | N/A | BV421 |
525/50 | 505LP | BV510, BV480 | |
610/20 | 595LP | BV605 | |
670/30 | 655LP | BV650 | |
710/50 | 690LP | BV711 | |
780/60 | 750LP | BV786 | |
Blue (488 nm) | 530/30 | 505LP | BB515, FITC |
610/20 | 595LP | BB630, PI | |
710/50 | 690LP | BB700, PerCP-Cy5.5 | |
Green-Yellow (561 nm) | 586/15 | N/A | BYG584, PE, TdTomato |
610/20 | 600LP | PE-CF594 | |
670/30 | 635LP | PE-Cy5, 7AAD | |
780/60 | 750LP | PE-Cy7 | |
Red (637 nm) | 670/30 | N/A | APC |
730/45 | 690LP | APC-R700 | |
780/60 | 750LP | APC-H7, APC-Cy7 |
Chromium 10x Genomics
It allows the isolation of dissociated cells in a droplet (up to 10,000 cells per sample) and the preparation of 3' libraries from the mRNA contained in each droplet. Sequencing of these libraries allows the study of single-cell gene expression (3' transcriptome) and thus the fine profiling of the tens of thousands of cells contained in the sample.
LASER | Filtres | Mirrors | Fluorochromes |
|---|---|---|---|
Violet (405 nm) | 448/45 | N/A | BV421, DAPI |
528/45 | 500LP | BV510, CFP, AmCyan | |
Blue (488 nm) | 488/15 | N/A | SSC |
527/32 | 507LP | FITC, GFP, BB515, AF488 | |
700/54 | 665LP | BB700, PerCP-Cy5.5, PerCP | |
Green-Yellow (561 nm) | 582/15 | 582LP | PE, DsRed |
613/18 | 605LP | PE-CF594, mCHERRY | |
697/58 | 665LP | PE-Cy5, PE-Cy.55 | |
783/56 | 752LP | PE-Cy7 |
- Chromium 10x Genomics. Cet appareil permet l’isolement de cellules dissociées dans une gouttelette (jusqu’à 10000 cellules par échantillon) et de préparer des librairies 3’ à partir des ARNm contenus dans chaque gouttelette. Le séquençage de ces librairies permet d’étudier l'expression génique unicellulaire (transcriptome 3') et ainsi de profiler finement les dizaines de milliers de cellules contenues dans l’échantillon.