Presentation

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The histology platform has currently all the equipment necessary to perform routine histological techniques on fixed, frozen or fresh samples, including the possibility of virtualizing your histological slides. The platform also has specific equipment for the realization of 3D marked sample reconstructions (OHREM) and laser microdissection projects.

 

Our mission is to take care of your samples according to your request (embedding tissues in paraffin, cutting tissue, histological standard or specific staining and scan of histological slide in bright field or in fluorescence, HREM) to train users on different histological techniques (embedding tissues in paraffin, immunohistochemistry) and on our various workstations available for reservation (microtome, cryostat, vibratome and laser microdissector).

 

We offer you our experience and technology for this platform.

The platform is open to external research teams from public and private institutions.

We will be happy to help you with specific and routine analyzes in your research projects.

 

1. For current service requests, please complete the dedicated forms and send them by email to histo.sfrnecker@inserm.fr

2. For all requests concerning the possibility of innovative technological developments or collaborative analyzes, please contact the manager of the platform Sophie Berissi by email sophie.berissi@inserm.fr (cc scientist referent Dr Ganna Panasyuk at ganna.panasyuk@inserm.fr)

Localisation 

Plateforme d’Histologie

1St Flour

Faculté de Médecine Université de Paris

156-160 rue de Vaugirard

75015 Paris

 

Tél : 01 40 61 54 80/81/85

histo.sfrnecker@inserm.fr

 

Staff

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Sophie Berissi

Head - IE INSERM

sophie.berissi@inserm.fr

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Ganna Panasyuk, PhD

Scientific referent​

ganna.panasyuk@inserm.fr

Sofian Ameur
Damien Conrozier
 

Equipement

Equipment for routine histological technics

Treatment of fixed tissues:

  • A fully enclosed vacuum tissue infiltration machinetissue processor (ASP300S, Leica) allowing automatic dehydration and paraffin impregnation to be carried out for up to 300 samples per day.

  • An inclusion plateembedding center (EC-350, Microm France) to embed the tissues with paraffin

  • A cassette dewaxer (TR 302, Tech-Inter) to remove excess paraffin around the completed block.

 

Sections:

 

  • Two semi-automatic microtomes (2 x HM 340E, Microm France) for performing paraffin cutting services, as well as a semi-automatic microtome (RM2145, Leica) made available forto users

  • A slide printer (SlideMate, Thermo Scientific) for writing slides

  • A cryostat (CM3050S Leica) for making frozen sections. This device is also accessible to autonomous users.

  • A vibratome (VT1200) for production making sections on fresh tissue. This equipment is also accessible to autonomous users.

 

Staining :

  • A multistainer (ST5020-CV5030, Leica) for performing histological standard or specific staining classic and specific tissue stains and for mounting with coverslip

  • A microscope (DME, Leica) allowing constant control of the quality of sections and colorationsstaining.

 

Scanner :

The platform has a slide scanner (Nanozoomer 2.0 HT, Hamamatsu). This system makes it able to produce virtual slides in bright field or in fluorescence. The virtualized blades slides can be used via with NDPview (Hamamatsu observation and annotation software).

Characteristics:

Objective 20x NA 0.75 with overall magnification 20X or 40X

Resolution:  0.23 μm / pixel (40 ×) and 0.46 μm / pixel (20 ×)

Equipment for specific techniques:

Laser microdissector

The platform has also a Laser Microdissector (Palm MicroBeam, Zeiss). It allows to study nucleic acids (DNA / RNA) and proteins of specific cell subpopulations.

Characteristics:

  • Brightfield imaging camera : AxioCam ICc1 rev.3 - CCD 1392x1040= 1,4 Mega Pixels

  • Fluorescence imaging camera: AxioCam MRm rev3 - CCD 1392x1040= 1,4 Mega PixelsFiltres : DAPI (EX 390-420, LP450), FITC/GFP (EX BP 450-490, EM BP 515-565), Rhodamine (EX BP 510-560, EM LP 590)

  • Objectives : 5x/0,25 ; 20x/0,4 ; 40x/0,6 ; 63x/0,75

 

OHREM:

The histology platform has an OHREM (Optical High Resolution Episcopic Microscope, Indigo Scientific). HREM is a 3D histology technique. It is useful for analyzing and quantifying the fine anatomy of thick and large samples and can also provide 3D LacZ and HRP reporter images from transgenic samples or ISH experiments. It is not compatible with fluorescent protractors, it does not achieve subcellular resolution; it is therefore complementary to confocal or light sheet microscopes, which are often limited in the size of samples that can be processed and are not compatible with non-fluorescent reporters. Useful link: https://dmdd.org

Characteristics:

  • 2 light sources: 585nm (enzymatic labeling) or 470nm (morphology)

  • Cutting thickness: 1 to 5µm

  • Sample size: up to 20mm

  • Resolution: 4 or 12 megapixel

  • Cutting time: 2 to 12 hours (depends on sample size and resolution).

  • A binocular magnifier (Stemi 508, Zeiss) for orientation of the samples during resin embedding for the HREM technique

        

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Fully enclosed tissue processor 

(ASP300S, Leica)

 

Inclusion plate

(EC-350, Microm France)

A cassette dewaxer

(TR 302, Tech-Inter)

Services

Routine services

The platform takes care of your samples and performs the services of your choice:

  • Impregnation and embedding of tissues in paraffin

  • Section: frozen tissue (5-30µm) and / or embedded-paraffin tissue (4µm) and / or on fresh tissue

  • histological staining: standard staining (Hematoxylin - Eosin, Toluidine blue) or specific staining of collagen (Masson's trichrome, Picro-sirius), polysaccharides (Periodic Acid Schiff)

  • the virtualization of slides in bright field and fluorescence.

  • Impregnation and embedding of tissues in resin for the section with the OHREM

  • the section of sample included in a resin and acquisition of images with the OHREM.

 
Training

The platform also provides training for one or more members of your team:

  • the use of microtomy equipment: microtome, cryostat and vibratome

  • the development of immunohistochemical labels

  • laser microdissection (training and support)

 
Make a service / training request

For any service and / or training request, you must contact us by email at histo.sfrnecker@inserm.fr

To access the various services, it is imperative to fully subscribe to the charter of use of the histology platform. A meeting is also offered to discuss your project, offer you the various services that meet your needs and explain how the histology platform works.

 
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  2. Phelep A, Laouari D, Bharti K, Burtin M, Tammaccaro S, Garbay S, Nguyen C, Vasseur F, Blanc T, Berissi S, Langa- Vives F, Fischer E, Druilhe A, Arnheiter H, Friedlander G, Pontoglio M, Terzi F. MITF-A controls branching morphogenesis and nephron endowment. PLOS Genet. 2017 Dec 14;13(12).

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  6. Brezillon N, Brunelle MN, Massinet H, Giang E, Lamant C, DaSilva L, Berissi S, Belghiti J, Hannoun L, Puerstinger G, Wimmer E, Neyts J, Hantz O, Soussan P, Morosan S, Kremsdorf D. Antiviral activity of Bay 41-4109 on hepatitis B virus in humanized Alb-uPA/SCID mice. PLoS One. 2011;6(12):e25096

  7. Alliouachene S, Tuttle RL, Boumard S, Lapointe T, Berissi S, Germain S, Jaubert F, Tosh D, Birnbaum MJ, Pende M. Constitutively active Akt1 expression in mouse pancreas requires S6 kinase 1 for insulinoma formation. J Clin Invest. 2008 Nov;118(11):3629-38.

 

Publications