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Cellular and Tissue Phenotyping Platform

The Cellular and Tissue Phenotyping Platform provides the entire scientific community with high-tech instrumentation.

Cécile Delarasse Principal Investigator
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Xavier Guillonneau Principal Investigator
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Luisa Riancho Technical manager
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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 
(exemples)

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 
(exemples)

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.

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