Equipment for routine histological technics

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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.

• 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).

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Characteristics:

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

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

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Equipment for specific techniques:

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Laser microdissector

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

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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:

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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

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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