The platform provides the Institute's teams with high-performance tools, expertise, technical assistance and a technology watch with the aim of supporting the research projects developed at the Institute with state-of-the-art technology.
The Platform aims to satisfy your requests in cytometry and analyses related to the research activities of the laboratories, and to be open to the entire academic (CNRS, INSERM, SU...) and industrial scientific community.
The Platform offers various services:
-provision of high-tech tools
-training of users
-Collaborations for research and development projects
-service provision
The staff makes every effort to ensure the smooth running of the platform:
-organization of the activity (booking schedule, data storage),
-training / assistance of users,
-monitoring of projects and analyses,
-maintenance and management of material and equipment.
These equipments are associated with service and technological development activities. The platform has, in fact, a role of training, collection and transmission of knowledge.
All these activities are carried out within the framework of services or the provision of tools.
Devices
Celesta SORP Becton Dickinson
Flow cytometry is a technique that allows high-speed analysis of a suspension of particles (cells, bacteria, parasites, balls ..), the individual characteristics of each of them such as the size, shape and complexity and any component or function that can be detected by a fluorescent compound. This cytometer does not allow cell sorting, however, depending on the applications, it may be carried out using magnetic beads.
Setup Celesta SORP
| LASER | LASER POWER | DETECTORS | FILTERS | MIRORS | EXEMPLES |
|
488nm FSC & SSC |
100 mW |
A B C |
780/60 695/40 530/30 |
750LP 670LP 505LP |
PerCP-Cy7 PerCP-Cy5.5 FITC, BB515 |
| 637nm | 100 mW |
A B C |
780/60 730/45 670/30 |
750LP 690LP |
APC-Cy7 APC-R700 APC |
| 405nm | 100 mW |
A B C D E F |
780/60 710/50 670/30 610/20 525/50 450/50 |
750LP 690LP 655LP 595LP 505LP |
BV786 BV711 BV650 BV605 BV510 BV421 |
| 561nm | 100 mW |
A B C D |
780/60 670/30 610/20 586/15 |
750LP 635LP 595LP |
PE-Cy7 PE-Cy5 PE-CF594 PE |
Applications
• Cell viability: 7AAD or Propidium Iodide staining
• Apoptosis assays: annexin V-7AAD or PI, subG1 peak
• Caspase activity
• Mitochondrial Membrane Potential JC-1 staining
• Cell proliferation, (CFSE , BrDU)
• Cell cycle
• Cytokines
• Antigen quantification
• Direct and indirect stainings, membrane or cytoplamic stainings
• Cell characterisation and phenotyping.......
Melody Becton Dickinson_Cytometer Cell Sorter
The Melody cell sorter is placed under PSMII
Setup Melody
| LASER | FILTERS | MIRORS | EXEMPLES |
|
488nm FSC & SSC |
488/15 527/32 700/54 |
507LP 665LP |
SSC FITC, GFP, BB515, Alexa Fluor488 PerCP, PerCP-Cy5.5, 7-AAD |
| 405nm |
528/45 |
448/45 500LP |
DAPI, Proliferation Dye 450, BV421 BD Horizon V500, BV510, AmCyan, CFP |
| 561nm |
582/15 613/18 697/58 783/56 |
582LP 605LP 665LP 752LP |
PE, DsRed mCherry, BD Horizon PE-CF594, PE-Texas Red, PI PE-Cy5, PE-Cy5.5 PE-Cy7 |
10X GENOMICS
The platform is equipped with the Chromium from 10x Genomics which 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.
Team
Scientific Manager Cellular Section
Email: cecile.delarasse@inserm.fr
Luisa Riancho
Technical manager
Email: luisa.riancho@inserm.fr
Xavier Guillonneau
Scientific Manager Molecular Section
Email: xavier.guillonneau@inserm.fr
Frédéric Blond
Bio-informatician Engineer
Email: frederic.blond@inserm.fr
Sebastien Augustin
Molecular Biology Engineer
Email: sebastien.augustin@inserm.fr
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