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Une plateforme orientée vers les détails et conçue pour répondre à tous vos besoins en histologie.

Une plateforme orientée vers les détails et conçue pour répondre à tous vos besoins en histologie


Sous la direction de notre pathologiste, la Dr Marie-Christine Guiot (MD), nos techniciens en histologie formés fournissent des services d'histotechnologie de la plus haute qualité aux utilisateurs de la communauté universitaire de McGill, ainsi qu'au secteur privé. Sous une surveillance stricte du contrôle qualité, vos échantillons sont traités rapidement.

Notre plateforme est équipée pour fournir des services histologiques généraux, y compris la section, et des services spécialisés tels que la coloration IHC, des colorations spécialisées personnalisées, les TMA et des solutions de pathologie numérique.

Notre mission est de fournir des services de haute qualité de manière efficace et rentable pour promouvoir votre recherche scientifique. Votre satisfaction est notre objectif le plus sincère.

Pour en savoir plus sur nos services, veuillez consulter la page suivante. Au besoin, n’hésitez pas à nous contacter par le biais du formulaire de contact plus bas.

Directeur : Dr. Luke McCaffrey
Co-directeur et pathologiste : Dr. Marie-Christine Guiot
Gestionnaire de la plateforme : Nicole Robinson, MSc
Technicien de recherche principal : Plinio Queiroz Da Cruz, BSc
Technicien de recherche : Joy Zhang, BSc

The Histology Platform has a diverse array of equipment, some is for staff use only, while other pieces of equipment are open for booking by trained users.

Staff use only

Tissue Tek VIP5 Vacuum Infiltration Processor:

An automatic, self-contained tissue processor, which holds up to 300 cassettes.  The VIP software is programmable for up to 20 different programs for use in the fixation, dehydration, clearing and paraffin of a variety of specimens.  We can customize programs to the specific needs of our clients.

Leica EG 1150H Embedding Center:

The Leica EG1150 modular tissue embedding center incorporates two separate components, the independent modules offer the flexibility to arrange embedding workflow for maximum efficiency. Tissues are oriented in a metal mold filled with hot paraffin.

Leica ST5020 Multistainer, Automatic Slide Stainer and Coverslipper:

The automatic Stainer produces consistent, high-quality results for both routine and special stains and can perform single or multiple protocols at the same time. Note, some specialty stains are performed manually.

Microtomes:

The Core has three microtomes for staff use. 1) Leica RM2255 automatic microtome 2) Fisher Microm HM340 semi-automatic microtome 3) Fisher Shandon Finesse ME automatic microtome.

Cryostat:

The Core has one cryostat for staff use. Fishers Cryostat Microm HM 525 features UV disinfection and maintains temperatures between -15 degrees Celsius and -35 degrees Celsius.
H4: Ventana Discovery Ultra:
The Core is equipped with a Roche Ventana Discovery Ultra, and automated IHC staining system. The Ventana has 30 individual and independent slide drawers, each allowing different experiments to occur in parallel. Both fully automated and semi-automated protocols can be developed within the Ventana to suit your needs.
The Ventana Discovery Ultra is only available for independent use by users at the GCI. All users must be trained by a Histology Platform Technician before they are able to use and book this machine. All non-GCI members are required to fill out a service request.
Features:
·    Steps that can occur within the machine: baking, deparaffinization, cell conditioning, staining, counter staining, and titration.
·     35 reagent position and 30 individual and independent slide drawers.
·     Enhanced temperature control.
 
H4: TMA Grand Master:
The TMA Grandmaster is a fully automated Tissue Microarrayer, able to generate TMA paraffin blocks. Please see below for machine specifics.
Block Capacity within Machine:
Highest capacity: 72 blocks; 60 donor blocks and 12 recipient blocks can be loaded at once. If more blocks are needed for the TMA, the blocks with be exchanged after the first set of punches is collected.
Core Diameters
 • 0.6 mm- maximum of 558 cores  
 • 1 mm- maximum of 286 cores  
 • 1.5 mm- maximum of 135 cores  
 • 2 mm- maximum of 84 cores
Fast microarrayer
 • 2 mm max. 84 cores
 • max. 12 seconds per core   
 • Simultaneous loading, imaging, drilling, and punching.
H4: Smart automation
Features automatic block height measurements to ensure the embedded cores are aliged with the recipient block surface, allowing all cores to be exposed during sectioning, without wasting valuable tissue.  
 In addition, the machine is capable of automatic barcode reading, donor block and label image saving for reference, project data saving into Excel file and Automatic PCR extraction.
H4: PCR Extraction
Extracted FFPE tissue samples are ready for DNA extraction and PCR analysis with commercially available kits. A clean block is used for this process to avoid cross contamination.

Either 6 PCR cassettes or 10 PCR tubes/cassettes can be used.

H4: Hamamatsu Nanozoomer S210:
The Nanozoomer S210 is a high throughput digital slide scanner. The S210 can quickly scan tissues and cells into high quality digital images perfect for publication, analysis, or collaboration.
Features:
·         Batch scanning: can scan up to 210 in one batch.
·         Has both 20x and 40x objectives
·         Performs fully automatic scanning.
·         Brightfield scanners: can image routine H&E slides, slides stained with a specialty stain or IHC slides.
·         Z-stack feature allows scanning of thick sections.
·         Free image viewing software, that works on both Mac and PC.
·         Image Management: slides can be downloaded remotely from SharePoint or can be downloaded onto an external hard drive.


H3: Equipment available for use

H4: Microtome and Cryostat:
The Core has a cryostat and a manual microtome station, including the microtome, a water bath, and an additional light, which are available for booking.
All users of the equipment must first be trained by Histology Platform Staff before being able to book or use the instruments.
To keep the equipment in good working order the following protocols must be followed:
1.    The equipment must be booked online.
2.   Before using the equipment, please sign into the logbook to keep track of usage, and note any errors or problems that arise.
3.  Users are required to bring their own tools to use the equipment (such as blades, slides or OCT).
4.    All equipment must be kept clean.
a.    Cryostat: Thoroughly clean the cryostat with 100% after use, including the mounts.
b.    Microtome: Must be cleaned after use, and the water bath emptied. Please turn off all pieces of equipment.
5.    Rummaging around Core drawers and taking core supplies without permission is not allowed. We are unable to quantify levels of our consumables if they are being taken without our knowledge or consent.
6.   If any issues should arise while using the equipment, please let Core staff know as soon as possible.
H4: Ventana Discovery Ultra and Nanozoomer S210:
The Ventana Discovery Ultra is open for use and booking by GCI members, please contact the Core for more information or to set up a training session.
The NanoZoomer S210 is currently for staff use only. In the future the Nanozoomer will be opened for GCI lab use, with labs able to select an individual to be the designated user for the lab.[NR1] 

 [NR1]Depends on when launching- will launch with infinity- no clue when that is 

 

How you submit your samples depends on what samples are being submitted. All samples must be submitted through the completion of histology request, which can be made on the Infinity Platform.

Paraffin Samples

All paraffin samples need to be fixed by the labs before they are dropped off with the Histology Platform. See FAQ section for fixation protocols.

All paraffin samples, whether they are in solution pre-processing, already embedded or sectioned, can be dropped off in the basket outside the Core labeled ‘Histology Drop Off’. All samples must be brought in with a printed copy of the request form generated from Infinity.  

All cassettes being dropped off at the Core should be labeled in pencil, solvent resistant markers or have been printed with a cassette printer.

If slides are being dropped off for staining, ensure that a sharpie is not used for labelling, as the staining process will remove the label. We recommend a pencil or an solvent-resistant marker.

If slides are being dropped off for scanning, please ensure they are completely dry and clean (no excess mounting media or broken glass).
Cassette Size: There are different sizes and types of cassettes one can use for samples, what is needed will depend on the size of your samples.

  • Mesh Cassettes: The openings in the cassette are mesh, making this the best option for small samples, such as salivary gland. If you have very small samples, it is advised they are wrapped in a lens paper before placing into the cassette, to ensure it does not fall out.
  • Standard Cassettes: Good for most samples. Can be used for small samples if wrapped in lens paper if mesh cassettes are not available.
  • Mega Cassette: Used in very large samples, or samples that are long and mush be embedded vertically. This could include whole rate femur or colon. 

Frozen Samples

All frozen requests require an appointment to be made with the Histology Core. Contact the Core at: histology.gcrc@mcgill.ca

On the day of your appointment, you will bring in your samples either on dry ice (fresh-frozen or already embedded in OCT) or at 4 degrees Celsius (if fixed and cryoprotected) to the Core with a copy of your request form. Technicians will indicate the length of time expected to complete your order so you can arrange pick-up. Samples will be stored short term at -20 or -80 if same day pick-up is not an option.

The Histology Innovation Platform, and all Platforms located at the Goodman Cancer Institute ask that you acknowledge the Core in your publications and presentations.

Acknowledging the Platform is Important

  1. Our clinicians and technicians offer valuable intellectual and experimental contributions to your research and deserve to be acknowledged for their time and effort.
  2. Our Platform requires proper acknowledgements in publications as a valuable metric. Acknowledgements are the most important way we can demonstrate our value and are extremely useful when submitting grants or requesting funding from various funding bodies to support the cost of services, or new equipment, which helps us gain financial support to maintain our operations. Without this support we may not be able to continue to operate or provide essential services to the research community.

How to Acknowledge the Platform:

  • When: The Platform should be acknowledged any time a Platform is used by your lab.
  • Where: Acknowledgements can be added to papers, publications, posters, grants, or reports
  • How: Here are some examples of how the Core can be acknowledged in your research:
    • Within the Acknowledgements section: The authors are grateful for support from the Histology Innovation Platform (Technicians names) at the Rosalind and Morris Goodman Cancer Institute.
    • Within the Acknowledgements section: Histological Experiments were performed by the Histology Innovation Platform (Technicians names) at the Rosalind and Morris Goodman Cancer Institute
    • Within the Methods Section: Experiment was performed by the Histology Innovation Platform at the Rosalind and Morris Goodman Cancer Institute
  • If there was a new stain or method developed by the Core for your research than technicians working on the project must be given co-authorship on your publications.

Please send us a PDF copy of each publication where the Histology Platform is acknowledged. This helps us to apply for funding to continue supporting researchers. Publications can be sent to the histology.gcrc@mcgill.ca email address

PROTOCOLS

Protocols will be added shortly.

Paraffin vs. Frozen Specimens

Paraffin embedded samples are the most stable samples and offer better preservation of tissue morphology and architecture with less injury to the tissue. Paraffin samples are more cost efficient and perform well with many different staining techniques including IHC.

Paraffin samples do have drawbacks including certain stains do not perform optimally with paraffin samples, DNA and RNA degradation can occur, and it is hard to extract nucleic acids for further downstream experiments.

There are two different types of frozen specimens: fresh frozen and cryoprotected tissue. Both frozen tissues will have increased lipid retention and improved preservations of enzymes and antigens. Often frozen sections are thicker than paraffin sections which can decrease the microscopic resolution and make it harder to study the tissue morphology.

Fresh frozen samples preserve the native biological state, thus you have more flexibility for downstream experiments, such as IHC with biomolecules that degrade when fixed. However, drawbacks include the continuous storage of samples at -80 degrees and lack of long-term preservation, gradual tissue degradation, the need for fast processing and sectioning.

Cryoprotected frozen, is first fixed and cryoprotected before being frozen. These steps aid in the tissues long term preservation, which is better for long term analysis of samples. Drawbacks can include the lack of cellular viability, long term storage at -80, and potential artifacts and alternations to the tissues.

For IHC experiments, what will the Platform need?

For IHC experiments you will need to fill out a request, and bring down either the blocks to be sectioned or slides to perform the experiment on.

You will also need to bring down the primary antibody for the Core to use. In most cases we will not have the antibody on hand. If you want to check with staff if your antibody is in stock, please send us an email.

If you are performing a duplex or triplex stain, you may need to bring down further reagents. Please contact us for more information.

When choosing a primary antibody for IHC, what do I look for?

Look for a primary antibody, choose one that either has anti-rabbit or anti-mouse as a secondary antibody.

Ensure the antibody has reactivity with your species of interest. This can be found on the antibodies data sheet.

You may choose a polyclonal or monoclonal antibody. Monoclonal antibodies are specific to a single epitope and will be able to detect a single protein with high affinity. A Polyclonal antibody will recognize multiple epitopes and is better suited for heterogenous populations.

Ideally choose an antibody that has already been tested and approved effective in IHC staining, this information can be found on the antibody manufacturer’s data sheet. In many cases you can see other publications that have been published using these antibodies, if deciding between antibodies, one can reference these publications to see if they have been used on your specific tissue or species.

What is a STEP Section?

STEP Sectioning is when a there is a space between sections collected. For example, if you desire a step of 100 microns, one sections will be taken and then the tissue will be trimmed, until a second section 100 microns away from the original section can be taken.

STEP section allows one to have sections from different areas within the same tissue.

What is the best way to ship samples to the Platform?

If samples are being shipped to the Histology Platform a date needs to be selected with the Core where your lab would arrange a courier service to pick up your samples and bring them to the Core. We will provide a detailed address and our lab phone number/ contact person so that we can receive the samples. The Core is open 9-4 on weekdays. Once the work is finished, your lab will send us a shipping label, and coordinate with the company to come and pick up the samples.

Please ensure that if your samples are frozen, that adequate measures are taken to ensure they remain frozen on the duration of their journey. Also ensure that all samples that have yet be processed or embedded (both FFPE and cryoprotected samples) are in enough solution to avoid dehydration.

Do we get to the keep the slide boxes or folders?

The Histology Platforms slide boxes need to be returned after collecting your slides or returned when dropping off your next set of samples. Boxes given out to labs are tracked by Core staff and marked when returned. Quarterly labs will be reminded of the missing slide boxes, and if not returned the labs will be charged.

You can bring down slide boxes for us to use with your order to simplify slide collection once the order is complete.

If you do not have slide boxes in your lab, you are able to request to buy ours as part of your order.

For IHC, do I need to provide control tissue?

We have tissue that can be used for controls in your IHC experiments performed at the Core. The Core has mouse, rat, and human control tissues, if you require a control tissue from another species, you will need to bring this to the Core for your IHC. Additionally, if a diseased state control is needed for your experiment, you will also need to provide this.

How do I know when my samples are ready?

When your samples are ready to be collected you will receive an email notification from the Core. You will be able to collect your samples from the pickup box, on the table outside the Core’s main entrance.

If you are picking up frozen samples, please come within the Cores working hours, as we will store samples in the freezer until they are picked up.

Additionally, if your slides were scan and you wish to download a copy onto a USB or external hard drive, please come during working hours so we can provide you with the necessary support.

How do you properly fix or prepare specimens?

Proper methods of fixation or sample preparation will depend on the type of sectioning being done. All fixation needs to be done quickly after euthanasia and dissection. Fixation allows for the preservation of the native structures of your tissues and is very important for the quality of the sections.

Paraffin: Fixation

Generally, tissues should be dissected into pieces that are between 3-5 mm thick. If larger tissues need to remain intact your fixation time may be longer. If working with small tissues, and cassetting yourself, ensure the samples are wrapped in lens paper to avoid the tissue falling out of the cassette.

To fix:

  1. After dissection, rinse tissue with PBS to remove blood or debris.
  2. Place tissue in fixative solution. The volume of fixative should be at least 15x to 20x greater than the volume of tissue.
    1. The most common fixative is 10% Neutral Buffered Formal (NFB).
  3. Samples are refrigerated at 4 degrees Celsius.
  4. Samples are left in fixative for approximately 24-48 hours, and usually no longer than 72 hours (if using NFB)
    1. This length of time may very if you have a very large tissue or are using a weaker/ stronger fixative.
  5. Samples are transfer into 70% Ethanol in a sealed container.
  6. Samples are ready to be brought to the Histology Platform- make sure to bring down a copy of the request form and label the sample container with the request number.

Frozen: Fixation and Cryoprotection

In cold conditions:

  1. Wash the tissue in cool PBS, after dissection.
  2. Fix in 10% NBF or 4% paraformaldehyde (can be done in Eppendorf or dish make sure volume of solution is about 20 times the volume of the tissue) Leave at 4 degrees for ~48 hours.
  3. Wash with cold PBS
  4. Place tissue in 15% Sucrose- PBS solution, 4 degrees, until the tissue sinks (usually overnight)
  5. Place tissue in 30% Sucrose PBS solution, 4 degrees, until it sinks (usually overnight)
  6. Send to Histology Platform on Ice, or ice packs in the Eppendorf tubes.

We will then be able to complete the OCT embedding and sectioning.

Frozen: Fresh Frozen

The tissue needs to be frozen very quickly, however directly submerging the tissue into liquid nitrogen can cause cracks which will affect the quality of your sections.

There are a few methods to fresh freezing.
Dry Ice:

  1. Place tissues into a tube or small container within a Styrofoam box with dry ice. This method may not be as quick or efficient at freezing.

Submersion

  1. Add dry ice into the bottom of a Styrofoam container and place a small disposable container or stainless-steel bowl onto the dry ice.
  2. Add some small dry ice pellets into the bowl.
  3. Pour isopentane/2-methylbutane into the bowl. The dry ice pellets and isopentane/2-methylbutane mixture will vigorously bubble.
  4. Once bubbling has stopped, place the tissue into the solution until it is completely frozen.
  5. Transfer the tissue into a container that has been pre-chilled on dry ice.
  6. Store the tissue in -80 or in liquid nitrogen.

 
For either method of fresh freezing, the samples must be brought down to core in dry ice to ensure integrity.

Specimen Orientation

At the Core our standard method is to embed so the largest surface area will be exposed when cutting. If you desire a specific orientation, or area of tissue to be exposed please write this in the note section of the request form, and you can attach an image to the request or an Excel file to give us a guide.

Section Thickness

Section thickness will depend on the downstream experiments being performed on samples and which instrument is being used.

For paraffin sections, 4 microns is the most common thickness, but section thickness can range from 2-10 microns.

For frozen sections, thickness usually ranges from 4-15 microns.

What objective do I choose for bright-field scanning?

There are two objectives you can choose from when scanning slides, either 20x or 40x.

20x is the most commonly used objective. These images allow one to see cellular features and structures. Additionally, the file sizes are still relatively small, making data management easier.

40x is used to give more detailed images. This can be important when looking at nuclear structures or even small bacteria within samples. 40x scanning produces large files, which if scanning many slides can make storing the data more complicated.

I need my samples ASAP, what do I do?

If you need your specimens done quickly for a tight deadline, you may select the RUSH option on the request. This prioritizes your submission and moves it to the front of the line. RUSH samples are guaranteed within 2 weeks- and are usually completed in under a week.

To note the RUSH option comes with an extra charge. RUSH requests add an additional 50% of the final order total onto the cost. If the request total cost is $10, the RUSH charge will be $5.

If I have used histology equipment before, do I still need training?

Yes. You still need training by a Histology Core staff member even if you have had experience with this type of equipment in the past. The training allows staff to ensure that you can use the equipment in a safe manner. Our priority is your safety and the maintenance of our equipment.

Le moyen le plus simple de trouver notre laboratoire est d'entrer par l'entrée principale du bâtiment Bellini (3649 Promenade Sir William Osler). Prenez l'ascenseur jusqu'au 3e étage et tournez à droite, passez devant la cuisine et vous devriez voir la Plateforme sur votre droite. La table de dépôt pour vos échantillons se trouve devant la porte. Si vous avez besoin de parler à l'un de nos techniciens, veuillez entrer dans le laboratoire. Veuillez noter qu'il est exceptionnellement difficile d'entendre quelqu'un frapper à la porte depuis la partie principale de notre laboratoire.

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