English

Screening

Contact : Benoît Bordignon

Project: co-financed under the Operational Program FEDER-FSE 2014-2020 Languedoc Roussillon

PERSONNEL

Technical manager
Screening /
Development
Bio-informatician
Statistics /
Image analysis
MGC Manager

Equipments

Pipeting robot:

Plates reader:

Robot:

  • Cell dispenser
CellDispenser
Fast liquid pipeting system, used for cell seeding (under sterile laminar flow hood).

Manual pipetting system:

Distributeur Manuel
Manual pipetting system for 96 wells at once (volume : 10 - 100 µl).

SpeedVac for multiwell plates:

Speed Vac
Speedvac for 2 plates.

Technology & expertise

  • High Content Screening


  • Our services

    Our screening service allows production of in vitro and in vivo screens for models compatible with 96/384 well plates. Our robotic equipment allows repetitive jobs to be performed in a repeatable and robust way, any project requiring automation can benefit from robotization of all or some of its steps. We are able to realize two types of screens, « high-throughput screening » (HTS) and « high-content screening » (HCS) depending on models, targets and desired parameters.

    We propose to accompany the project leaders in each step of a screen. First, we help to optimize a fonctional test protocole, in 96-well plate format, on the projet targets. Secondly, we transfer a manuel protocol to robot (automation), we carry out a series of preliminary tests with spécific controls (positive and negative) to evaluate robustness and reproducibility of the automated protocol. In parallel, we optimize acquisition and analysis methods adapted to sreen parameters. Finally, we perform the complete screening and proceed to a statistical analyzes of the results allowing "Hits" identification.

     

    High-Throughput Screening (HTS)

    For a HTS-type screen, we use the TECAN FreedomEVO 200 platform. It contains two pipetting arms with 8- and 96-channels, a 96/384 wells plate washer, a carousel and a high capacity CO2 incubator, as well as a multimode microplate reader up to 1536 wells format. This platform allows the realization of fully automated HTS screens, from preparation of samples to the final reading of plates, through various treatments and incubations.

    High Content Screening (HCS)

    For a HCS-type screen, we also use the TECAN FreedomEVO 200 platform for automated sample preparation. We then use the ThermoScientific Cellomics (ArrayScan) for the HCS. It is an instrument dedicated to acquisition, measurement and multiparametric analysis of cellular content for a large number of samples. It contains an automated microscope (Carl Zeiss) for the analysis of fixed or living samples in 96-well (and 384) plate formats. It offers very wide analytical possibilities adapted to various fields (immunostaining, cytotoxicity, pharmaco-kinetics, oncology, cell biology, etc.).

     

    Terms of Access

    Contact us for any information on screening, for a quotation or for implementation of a collaborative project.

    Indicative prices ranges (decreasing depending on volume):

          - HTS screening of Chimical Library like Prestwick, 1280pts: € 3,000 - € 5,000
          - HCS screening with Immunolabelling (IF) and Imaging,1280pts : € 4,000 - € 7,000
          - HCS screening for siRNA library, 2300pts: € 5,000
          - Low-throughput
    assay as IC50 (5 drugs; n=3): € 2,000 - € 3,000

     

    Our banks

    ORFEOME via MGC Platform

    website: https://www.biocampus.cnrs.fr/index.php/en/facilities-en/browse-by-facility/platform/366-mgc-en

    https://www.igmm.cnrs.fr/en/service/montpellier-genomic-collection-mgc/

     

    siRNA Humans Libraries: Kinesins/Myosins/Dyneins; Membrane Trafficking; Rho GEFs

    website: https://horizondiscovery.com/products/screening/screening-libraries/Gene-modulation-screening-libraries/siRNA/human-sirna-libraries

     

    Crispr Cas9: Sanger Human whole genome Crispr libraries, lentiviral and glycerol stock

    website: https://www.sigmaaldrich.com/life-science/functional-genomics-and-rnai/sanger-crispr-arrayed-library.html

     

    Essential Compound Library, a representative sample of the French National Chemical Library (CNE : 1000 compounds)

    website: http://chimiotheque-nationale.cn.cnrs.fr/?Presentation

    https://chembiofrance.cn.cnrs.fr/en/

     

    Anti-cancer Compound Library (4730 compounds) TargetMol

    site web : https://www.targetmol.com/compound-library/anti_cancer_compound_library

     

    Prestwick Chemical Library, update 2019 (1520 compounds)

    website: https://www.prestwickchemical.com/screening-libraries/prestwick-chemical-library/

     

    Lopac 1280 Pharmacologically Active Compounds Library

    website: https://www.sigmaaldrich.com/FR/en/product/sigma/lo4200?context=product

     

                                                  

    How to find us?

    CRBM UMR5237

    Rez-de-chaussée porte 141,
    1919 route de Mende
    34293 Montpellier Cedex 05

    Téléphone : +33 4 34 35 95 18

    GPS : Latitude = 43.63767 ; Longitude = 3.86544

    SEM volutome zeiss

    
    Tarif établissement partenaire: -- €/heure
    Tarif établissement public et privé : -- €/heure

    Brève description :
    Gemini 360 Zeiss équiped with a Volutome (serial block face)
    Optimized for high resolution imaging of biological samples
    Mode High Vacuum (HV)
    Mode Variable Pressure (VP) up to 60 Pa
    Focal Charge Compensation (FCC) with the Volutome
    Max resolution (x,y) en HV:
    - 0,7 nm at 15kV
    - 1.2 nm at 1kV
    - 0.6 nm in STEM at 30kV
    Detectors:
    - SE2 (Everhart-Thornley): secondary electrons in chamber HV
    - C2D (à casacde de courant): électrons secondaires en chambre VP
    - BSD Volutome: backscattered electrons, retractable,high resolution low voltage
    - in Lens SE et BSD
    - aSTEM4: transmitted electons, retractable, 4 segmenys
    Sample holders
    - Standard holder 9 stubs
    - Correlative CorrMic holder for coverslips; compatible with Zeiss light microscope
    - STEM holder for 12 standard grids
    - Volutome sample holder
    Softwares:
    SMARTSEM : microscope control
    ZEN Core : volutome
    Atlas 5: automated acquisition 2D (mosaïcs, multiposition, multiscale)
    Atlas array tomography: automated acquisition of serial sections
    Zeiss Volutome
    Maximal volume : 1x1x1 mm
    Minimal section thickness: 20 nm (=max Z resolution max)
    

    SPIM: Lightsheet Z.1

    
    Partner lab price : 15.66 €/hour
    Public and private lab price: 55.41 €/hour
    Associated technologies:
  • 3D imaging
  • Videomicroscopy
  • In vivo imaging

  • Brief description:
    Light Sheet microscope Z.1 Zeiss for fast acquisitions of live samll organisms like embryons ou cleared samples. The microscope is equipped with one illumination arm and one detection arm with one 20X objectives and 2 cameras.

    Controlled by Zen software
    Objectives:
    20X / water n=1.45
    20X clearing objective
    5X clearing
    Détectors:
    PCO.Edge, sCMOS, 1920 × 1920 pixels

    Illumination sources:
    laser 405nm
    laser 488nm
    laser 561nm
    Spectral selection:
    4 dual band emission filters in a filter wheel
    DAPI-GFP
    DAPI-Cy3
    GFP-Cy3
    GFP-DRAQ5

    Incubation:
    temperature between 10°C and 40°C - CO2 control possible
    2 incubation chambers: a classical one for fixed or live samples and one for cleared sample
    Acquisition mode
    -multi-dimensional light-sheet acquisitions: multi-channels, multi-Z stacks, multi-timepoints, multi-angles with rotation of the sample up to 360°
    Sample carriers:
    the sample is prepared in a glass capillary either embedded in a gel or loaded in a well formed in the gel. This capillary is charged in the sample carrier on the stage.
    Each project done on this Light Sheet needs specific development for the sample preparation

    Spinning disc Nikon Ti Andor CSU-W1

    
    Partner lab price : 15.66 €/hour
    Public and private lab price: 55.41 €/hour
    Associated technologies:
  • 3D imaging
  • Videomicroscopy
  • In vivo imaging

  • Brief description:
    ANDOR confocal spinning disk on an inverted NIKON microscope (Ti Eclipse)

    Dedicated for live samples observations on multiweel or petridishes

    XYZ motorized microscope and temperature and CO2 control, focus control

    B&W camera Zyla 4.2, software : iQ3 by Andor
    Objectives :
    - 4x/0.13
    - 10x/0.45
    - 20x/0.75 DIC
    - 40x/1.15 Water
    - 60x/1.4 Oil DIC
    - 100x/1.45 Oil DIC

    Fluorescence filters :
    - DAPI ex BP377/50, DM409
    - FITC ex BP482/35, DM506, Em BP536/40
    - Texas Red ex BP562/40, DM593, Em BP624/40
    - Cy5 ex BP628/40, DM660, Em BP692/4

    Lasers :
    - 405
    - 488
    - 561
    - 640

    QUAD filters for emission :
    425-470nm - 505-542nm - 578-623nm - 660-750n
    Spinning disk Yokogawa head CSU-W1 equiped with two dichroïcs (3 bands 405/488/561 and 4 bands 405/488/561/640)

    B&W Caméra Zyla 4.2
    field = 2048x2048pxl, pixel size = 6.5µm
    IQ3 software
    Motorized microscope in XYZ : 3D stack, timelapse with variable frequency, multiposition, mosaïque, multichannel (combined dimension for handmade experimentations)

    Ideal operating temperature between 19 and 26 °C

    L2 Containment

    Spinning disk Dragonfly

    
    Partner lab price : 17.47 €/hour
    Public and private lab price: 82.24 €/hour
    Associated technologies:
  • 3D imaging
  • Videomicroscopy

  • Brief description:
    Nikon inverted micorsopce coupled to the Andor Dragonfly spinning disk dedicated to 3D confocal imaging of live samples or fixed samples. This microscope is also equiped 2 cameras, for simultaneous 2 color live imaging.

    Controlled by the Andor fusion software.
    Objectives:
    10X Plan Apo lambda 0.45 NA 4mm WD dry
    20X Plan Apo lambda 0.75 NA 1mm WD oil
    40X Plan Fluor 1.3 NA 0.2mm WD oil
    40X Apo 1.15 NA 0.6mm WD water
    60X Plan Apo lambda 1.4 NA 0.13mm WD oil
    100X Plan Apo lambda 1.45 NA 0.13mm WD oil

    Detector:
    2 camaras EMCCD iXon888 Life Andor
    (1024*1024 pixels, 13µm pixel size, 2x lens if needed, QEmax 95%)

    Environmental control: temperature and CO2
    Illumination sources:
    1 laser bench for confocal mode:
    405nm (100mW), 488nm (150mW), 561nm (100mW) et 637nm (140mW)

    Spectral configuration:
    2 dichroïcs in the head unit: either 100% to one camera or DAPI/GFP for vertical camera and mCherry/Cy5 for other camera with 565 dichroic mirror
    6 emission filters in the external wheel:
    DAPI (450/50), GFP (525/50), mCherry (620/60), Cy5 (700/775), GFP/mCher
    Type of acquisition:
    multi-dimensional acquisition in bright field or DIC transmission and confocal in fluorescence: XYZ (Z piezo of range 300um), multi-channels, multi-position, tile-scans, multi-wells, timelapse
    hardware autofocus (PFS)

    Compatible sample holders:
    35mm dish with #1.5 glass bottom, glass slide with #1.5 glass coverslip or glass-bottom multi-well plate

    Spinning disk Nikon Ti Inscoper CSU-X1

    
    Partner lab price : 15.66 €/hour
    Public and private lab price: 55.41 €/hour
    Associated technologies:
  • 3D imaging
  • Videomicroscopy

  • Brief description:
    Nikon inverted TiE micorsopce coupled to a spinning Yokogawa CSU-X1 unit dedicated to 3D confocal imaging of live samples. All the peripheriques are controled by the Inscoper electronic box, that increases the acquisition speed (triggering).

    Controlled by the Inscoper software
    Objectives:
    10X Plan 0.25 NA dry
    20X Plan Apo lambda 0.75 NA dry
    40X Plan Fluor 1.3 NA oil
    60X Plan Apo lambda 1.4 NA oil
    100X Plan Apo lambda 1.45 NA oil

    Detector:
    1 sCMOS back-illuminated PRime95B Photometrics
    (1200*1200 pixels, 11µm pixel size)

    Environmental control: temperature and CO2
    Illumination sources:
    1 laser bench Oxxius:
    405nm (50mW), 488nm (100mW), 561nm (100mW) and 638nm (100mW)

    Spectral configuration:
    3 dichroïcs in the head unit: MONO (488), DUAL (488-561) and QUAD (405-488-561-640)
    6 emission filters in the external wheel:
    DAPI(447/60), GFP(525/30), RFP(607/36), Cy5(685/40), GFP/Cherry(512-23/630-90), DAPI/GFP/Cherry/Cy5 (440-40/521-21/607-34/700-45)
    Type of acquisition:
    multi-dimensional acquisition in bright field or DIC transmission and confocal in fluorescence: XYZ, multi-channels, multi-position, mosaïcs, multi-wells, timelapse
    hardware autofocus
    custom-made protocol thank to Inscoper software
    Compatible sample carriers:
    35mm dish with #1.5 glass bottom, glass slide with #1.5 glass coverslip or glass-bottom multi-well plate

    Spinning disk OLYMPUS SR

    
    Partner lab price : 17.47 €/hour
    Public and private lab price: 82.24 €/hour

    Brief description:
    Olympus IX83 inverted microscope coupled to Yokogawa W1 spinning disk unit, including a SoRa disk, dedicated to 3D confocal, high resolution imaging of live samples or fixed samples. A FRAP photoactivation module is coupled to the microscope.

    Controlled by CellSens software.
    Objectives:
    10X UPLSAPO 0.40 NA 3.1mm WD air
    20X UPLSAPO 0.8 NA 0.6mm WD air
    40X UPLXAPO 1.4 NA WD 0.13mm oil
    60X UPLXAPO 1.42 NA WD 0.15mm oil
    30X UPLSAPO 1.05 NA WD 0.73mm silicon
    60X UPLSAPO 1.3 NA WD 0.3mm silicon


    Detector:
    camera sCMOS Fusion BT Hamamatsu
    (2304*2304 pixels, 6.5µm pixel size, QEmax 95%)

    Environmental control: temperature and CO2
    Illumination sources:
    1 laser bench for confocal mode:
    405nm (50mW), 488nm (100mW), 561nm (100mW) and 640nm (100mW)
    1 laser bench for FRAP
    405nm (50mW), 488nm (100mW)

    Spectral configuration:
    8 emission filters in the external wheel:
    DAPI (447/50), GFP (525/50), GFP narrow (520/10), mCherry (593/40), TexasRed (617/73), Cy5 (685/40), Dual GFP-TexasRed (527-645), QUAD (435-520-595-695)
    Type of acquisition:
    multi-dimensional acquisition in bright field and confocal in fluorescence: XYZ (Z piezo of range 300μm), multi-channels, multi-position, tile-scans, multi-wells, timelapse
    hardware z drift compensation (ZDC2)
    Photoactivation FRAP module (405, 488nm laser)

    Compatible sample holders:
    35mm dish with #1.5 glass bottom, glass slide with #1.5 glass coverslip or glass-bottom multi-well plate