Dehydration and embedding paraffin automaton
A paraffin dehydration and impregnation automaton, Logos One, Thermo Fischer, MM France, acquired in 2017, used only by platform personnel allows automatic dehydration and impregnation in paraffin of 210 samples at a time, whatever the tissue type or species.
The protocol is a long cycle with an increasing ethanol dehydration step followed by isopropanol followed by the paraffin impregnation step.
The block inclusion is performed the next morning manually on a Tissue -Tek inclusion table.
Cryostat and Microtomes
The platform has different cutting equipments:
- 1 cryostat, NX 70, Thermo Fischer, MM France, acquired in 2018, for sectionning of frozen tissues. Recommended thickness of sections: 7µm
- 1 cryostat, Leica, on L2 for the treatment oh human tissues
- 1 microtome, RM 2155, Leica, for slices with a recommended thickness of 4µm, from fixed paraffin-included tissues
- 1 microtome, RM 2145, Leica, for users
- 1 fully automated microtome, Autosection, Tissue Tek, Sakura, acquired in 2018, for the exclusive use of platform staff, for routine sections but also for more difficult samples. This microtome automatically and systematically performs the parallelism between the paraffin tissue block and the blade to limit the loss of tissue material.
Slides Austostainer
An autostainer, SPECTRA, Leica, acquired in 2019. The staining automation, for both frozen and embedded paraffin tissues, offers the possibility to integrate 30 slides independently every 5 minutes and to carry out standard stainings (HES, HE, Trichrome de Masson or specific stainings -Red Sirius (collagen and fibrosis), Pearls (iron), Oil-red-O (lipids and steatosis), PAS (Schiff Periodic Acid) and Blue Alcian (mucopolysaccharides)…
Dewax protocol and Dehydration protocol are also available for manually treated slides between these 2 steps.
Immunostaining automaton
An unmasking chamber (Abcys) essential for the antigen retrieval in pre-treatment of manual fluorescence or brightfield immunostainings, for FFPE (fixed formaldehyd and paraffin embedded) tissues or frozen fixed tissues . It allows a controlled, regulated and effective unmasking of antigenic sites from tissue sections in order to make them accessible to primary antibodies.
A Bond RX (Leica) immunostaining automaton for the management of all types of immunolabeling for time saving and improved quality, reliability and reproducibility of results. Its capacity is 30 slides, all independent. It is for the exclusive use of the platform staff for technical services.
Different types of immunostainings are possible:
- Single Fluorescence
- Single Chromogenic (DAB/ Brown or Red)
- Double or triple fluorescence staining
- Double brightfield immunostaining ( Brown and one other chromogenic)
- OPAL Technology Multiplexing (In Development)
- Ultivue Technology Multiplexing (In development).
Tissue Micro Arrayer
The TMA System, Minicore, Excilone, acquired in 2018, is a semi-automated system that allows the creation of Tissue Micro Array blocks. From dozens of donor paraffin blocks, we obtain a recipient paraffin block composed of a sampling of dozens of different tissues. It is therefore a question of reducing the number of blocks to be processed in order to save time, costs and reagents. The different tissues are then all treated on a unique slide, at the same time, under identical experimental conditions for a better reliability of the results and an easier analysis of the results.
Slide scanner
A slide scanner, the "Lamina slide scanner" (250 automatic slides) from Akoya Perkin Elmer. This automaton is able to acquire your entire slides in few minutes, whether in brightfield (histological stainings and brightfield immunostaining) or fluorescence, and to creat high-resolution virtual slides. Virtual slides are then very easily observable with specific software, such as Case Viewer or Panoramic Viewer, which can be downloaded free of charge from the internet. With these softwares called «virtual microscopes», you can zoom in on the regions of interest without losing any resolution, make captures, annotations, measurements, manual counts.
Below, the excitation and emission wavelengths of the available filters:
|
Filters |
Excitation |
Emission |
Canal |
|
DAPI |
387 ± 18 |
440 ± 44 |
1 |
|
FITC |
485 ± 26 |
521 ± 27 |
2 |
|
TRITC/TsRed |
559 ± 32 |
607 ± 40 |
3 |
|
Cy5 |
649 ± 19 |
700 ± 57 |
4 |
|
TRITC-BZ-HE |
543 ± 20 |
593 ± 40 |
5 |
Laser microdissector
An Arcturus XT laser microdissector, Excilone. The laser microdissection technique makes it possible to eliminate the heterogeneity of a tissue by dissociating, directly on tissue sections, with IR or UV lasers, different cellular types or territories of interest, to analyze them at the molecular level, specifically, independently of the tissue environment and thus, also, reveal weak molecular expressions. This technique is mainly recommended on frozen sections, previously stained with cresyl violet or immunostained (brightfield or fluorescence). The collection of the cell population or region of interest is done using a capsule that will then be adapted to a PCR tube containing the lysis buffer for the extraction of proteins, DNA or RNA.
Available Filters for immunofluorescence:
|
Filters |
Excitation |
Emission |
|
DAPI |
387 ± 18 |
440 ± 44 |
|
FITC |
450-490 |
520 |
|
TRITC |
540/25 |
605/55 |
