HISTOLOGY
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)
SERVICES
ROUTINE SERVICES
The platform takes care of your samples and performs the services of your choice:
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Impregnation and embedding of tissues in paraffin
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Section: frozen tissue (5-30µm) and / or embedded-paraffin tissue (4µm)
and / or on fresh tissue
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histological staining: standard staining (Hematoxylin - Eosin, Toluidine blue)
or specific staining of collagen (Masson's trichrome, Picro-sirius), polysaccharides (Periodic Acid Schiff)
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the virtualization of slides in bright field and fluorescence.
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Impregnation and embedding of tissues in resin for the section with the OHREM
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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:
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the use of microtomy equipment: microtome, cryostat and vibratome
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the development of immunohistochemical labels
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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.
EQUIPMENT
Equipment for routine histological technics
TREATMENT OF FIXED TISSUES
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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.
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An inclusion plateembedding center (EC-350, Microm France) to embed the tissues
with paraffin
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A cassette dewaxer (TR 302, Tech-Inter) to remove excess paraffin around
the completed block.
SECTIONS
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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
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A slide printer (SlideMate, Thermo Scientific) for writing slides
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A cryostat (CM3050S Leica) for making frozen sections. This device is also accessible
to autonomous users.
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A vibratome (VT1200) for production making sections on fresh tissue. This equipment
is also accessible to autonomous users.
STAINING
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A multistainer (ST5020-CV5030, Leica) for performing histological standard or specific staining classic and specific tissue stains and for mounting with coverslip
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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:
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Objective 20x NA 0.75 with overall magnification 20X or 40X
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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:
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Brightfield imaging camera : AxioCam ICc1 rev.3 - CCD 1392x1040= 1,4 Mega Pixels
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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)
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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:
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2 light sources: 585nm (enzymatic labeling) or 470nm (morphology)
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Cutting thickness: 1 to 5µm
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Sample size: up to 20mm
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Resolution: 4 or 12 megapixel
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Cutting time: 2 to 12 hours (depends on sample size and resolution).
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A binocular magnifier (Stemi 508, Zeiss) for orientation of the samples during resin embedding for the HREM technique
STAFF
Sophie BERISSI
Head
Sofian AMEUR
Mayeul THOMAS
Ganna PANASYUK
Scientific Referent
Damien CONROZIER
PUBLICATIONS
Haemodynamic stress-induced breaches of the arterial intima trigger inflammation
and drive atherogenesis.
Franck G, Even G, Gautier A, Salinas M, Loste A, Procopio E, Gaston AT, Morvan M,
Dupont S, Deschildre C, Berissi S, Laschet J, Nataf P, Nicoletti A, Michel JB, Caligiuri G.
Eur Heart J. 2018 Dec 11.
MITF-A controls branching morphogenesis and nephron endowment.
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.PLOS Genet. 2017 Dec 14;13(12).
Alternative splicing-regulated protein of hepatitis B virus hacks the TNF --
stimulated signaling pathways and limits the extent of liver inflammation.
Pol JG, Lekbaby B, Redelsperger F, Klamer S, Mandouri Y, Ahodantin J, Bieche I,
Lefevre M, Souque P, Charneau P, Gadessaud N, Kremsdorf D,Soussan P.
FASEB J. 2015 May;29(5):1879-89.
Cystic gene dosage influences kidney lesions after nephron reduction.
Canaud G, Bienaimé F, Viau A, Treins C, Baron W, Nguyen C, Burtin M, Berissi S,Giannakakis K, Muda AO, Zschiedrich S, Huber TB, Friedlander G, Legendre C, Pontoglio M, Pende M, Terzi F.Nat Med. 2013 Oct;19(10):1288-96.
AKT2 is essential to maintain podocyte viability and function during chronic
kidney disease.
Canaud G, Bienaimé F, Viau A, Treins C, Baron W, Nguyen C, Burtin M, Berissi S,
Giannakakis K, Muda AO, Zschiedrich S, Huber TB, Friedlander G, Legendre C, Pontoglio M, Pende M, Terzi F.
Nat Med. 2013 Oct;19(10):1288-96.
Antiviral activity of Bay 41-4109 on hepatitis B virus in humanized Alb-uPA/SCID mice.
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. PLoS One. 2011;6(12):e25096
Constitutively active Akt1 expression in mouse pancreas requires S6 kinase 1
for insulinoma formation.
Alliouachene S, Tuttle RL, Boumard S, Lapointe T, Berissi S, Germain S, Jaubert F, Tosh D, Birnbaum MJ, Pende M.
J Clin Invest. 2008 Nov;118(11):3629-38.