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Xylobiotech technical facilities : forest biotechnology
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INRA Val de Loire facilities (Orléans)

INRA, Centre Val de Loire - Orléans, 2163, Avenue de la pomme de pin, CS 4001 Ardon, 45075 Orléans cedex 2

  • Tel: +33 (0)2 38 41 78 24 (Accueil, P. Montes)
  • F. Laurans, responsable INRA activités XYLOBIOTECH
  • M.C. Lesage-Descauses, responsable INRA activités XYLOBIOTECH
  • Fax: +33 (0)2 38 41 48 09
  • Email:

General presentation:


The INRA site of XYLOBIOTECH is providing to users a confocal laser scanning microscope Zeiss LSM 700, protocols for the preparation of plant samples as well as an expertise in plant cell anatomy.


This equipment together allowed the observation of i) multiple fluorescence associated to colocalization analysis, ii) the 3D and iii) spectral imaging. It also offers possibilities for imaging the living plant cells (FRAP, FLIP, photo activation and conversion) to study intracellular trafficking.

The microscope is localized at the histocytology lab of the Research Unit AGPF (Forest Improvement, Genetics and Physiology) dedicated to the study of wood formation. The lab has a recognized expertise in histology and microscopic observation adapted to the study of wood formation.

Responsible Engineer for the INRA technical facility of XYLOBIOTECH :


Françoise Laurans  (IR2 INRA - UR AGPF)
+33 (0)2 38 41 78 38
Email:

Access to the technical facility:

Anyone interested in using a XYLOBIOTECH equipment will require the expertise of the responsible engineer of the technical facility at INRA Orléans who is in charge of the technical management. The responsible engineer will help in designing experiments and will give any advice and practical support for sample preparation and other experimental issues.

Any putative applicant may first contact the responsible engineer to discuss the objectives and conditions of the XYLOBIOTECH service. User’s needs in the frame of technical capacities of the XYLOBIOTECH equipment will be then formalized within a request form. A specific quotation will be written for each user’s demand and management fees will be charged.

Access to the technical facility from Monday to Thursday (8:30am – 6:00pm).

Equipment description :

Confocal laser scanning microscope

Microscope   
Zeiss LSM 700
Statif   
Right Axio Imager Z2 on antivibration table
Laser scans Laser diodes 405 nm, 488 nm,  555 nm, 639 nm
Filters   
DAPI, GFP, CY3/Rhodamine
Detectors   
2 PMT detectors (reflection), 1 detector (transmission)
Objectives   
10x/0,3 M27 Plan-Neofluar
20x/0,8 M27 Plan-Apochromat coulisseau DIC
40x/1,0DICM27 Plan-Apochromat
63x/1,40 oil DIC M27 Plan-Apochromat
Light sources Fluorescence Metal halide (120W), bright light halogen 100W
XY course Piezo-motorized deck & Joystick
Z course Motorized with accuracy of 10 nm
Speed acquisition 5 images/second
Imaging acquisition
2D/3D imaging (multi-probes)
Simultaneous or sequential acquisition
Spectral separation
FRAP (FLIP)
FRET
Imaging acquisition software
ZEN 2010


Some examples of services and associated costs:

************************Example 1************************

Colocalization of poplar wood fibers in cell walls after immunostaining on semi-thin wood tissue slices (low-definition image associated to the web site).

Immuno-localization of two classes of cell wall components in wood fibers using confocal laser scanning microscopy.

A Initial transmission imaging
B Colocalization of the 2 antibodies
C Localization of the first antibody labeled with Alexa 488
D Localization of the second antibody labeled with Alexa 633

Bar scale: 20 µm

Number of semi-thin wood tissue slices observed: 24
Timing for observation: 8 heures
Cost : 400 € without VAT

Cost estimation included observation fees. It did not include the production of wood tissue slices and immunostaining experiment.

************************Example 2************************

Microphenotyping of transgenic poplar plants produced though multigene co-transformation and modified for lignin metabolism (low-definition image associated to the web site).
Xylem analysis at the microscopic scale highlighted the strong autofluorescence of cell walls located around vessels.

Autofluorescence of xylem cell walls in wild phenotype (A, B, and C) and transgenic line (D, E, and F) observed with confocal laser scanning microscope (excitation 488 nm, LP filter 490 nm). A and E : transmission imaging.

Bar scale: 50 µm

Number of semi-thin wood tissue slices observed: 18
Timing for observation: 4 heures
Cost : 200 € without VAT

Cost estimation only included observation fees. It did not include the production of wood tissue slices.



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