The mission of the histology facility at Sunnybrook Research Institute (SRI) is to meet the expanding research needs of basic scientists and clinical researchers who require tissue processing. Our facility offers both standard and tailored research-specific histology services, catering to a diverse range of tissue samples and employing various staining techniques based on specific research objectives. We are equipped with an automated tissue processor, embedding station, motorized microtomes, autostainer, coverslipper, cryostats and a vibratome that allow tissue processing to be carried out rapidly and reproducibly.
The histology facility offers:
- Full-service processing of tissue samples. Services include consultations, tissue processing, decalcification, paraffin embedding, paraffin sectioning, cryosectioning, vibratome sectioning,routine and special staining.
- Training on histological equipment.
- Do-it-yourself access to histological equipment.
For a full listing of equipment, see the Equipment section. For services and fees, see the Services section and Fees section. Services are available to the SRI community and to investigators from external institutions.
Equipment
The Leica Autostainer XL is used for hematoxylin and eosin staining.
The Leica EG 1160 embedding station embeds specimens in paraffin blocks.
The Leica VT1000 S vibrating blade microtome can section fixed or fresh specimens immersed in buffer solution.
| Service/Equipment | Internal Fees |
|---|---|
| Paraffin | |
| Processing/embedding | $10.50/block |
| Multiple tissues processing/embedding (more than two per cassette) | $12/block |
| Microtomy (first slide) | $6/block |
| Cut extra sections from same tissue cassette) | $3.75/slide |
| Cut sections from (different) pre-existing block | $3.75/slide |
| Hematoxylin and eosin (H&E) staining | $3.75/slide |
| Special handling fee * | $60/hour |
| Frozen | |
| Cut section from block onto one slide | $3.75/slide |
| Collect additional level(s) on same slide (price per additional level | $2.63/section |
| H&E staining of section (from frozen tissue block) | $3.75/slide |
| Special handling fee * | $60/hour |
| Vibratome | |
| Handling fee (not embedding in agarose) | $7.50/block |
| Handling fee (embedding in agarose) | $15/block |
| Section (for histology) | $1.50/section |
| Special handling fee * | $60/hour |
*Special handling fee is applied additionally depending on a request or when it requires an extensive time to produce a single slide.
**Special staining is available upon request
***External researchers please contact Petia for pricing.
Tissue for paraffin embedding
For optimal preservation of tissue morphology, it is recommended that the thickness of the tissue fall within the range of 0.2 cm to 0.5 cm.
- For subsequent hematoxylin and eosin (H&E) staining, it is advisable to utilize a fresh solution of 10% neutral buffered formalin, ensuring a volume at least 10 times that of the specimen.
- If further immunohistochemistry is planned, we suggest reviewing the datasheet of the primary antibody for information on any optional fixative types and the recommended duration of fixation.
Tissue for frozen sectioning (cryosectioning)
Un-fixed and fixed tissue samples can be frozen sectioned.
- For unfixed tissue, it is crucial to freeze it as quickly as possible after harvesting to prevent the damaging effects of autolysis.
- In the case of fixed tissue, formalin-fixed tissue can be cryopreserved by immersing it in a 30% sucrose solution. This process helps protect cellular structures and components from damage during freezing by reducing the formation of ice crystals within the cells. Finally, the specimen can be embedded in OCT and stored as a frozen block.
For help with freezing technique, contact Petia Stefanova at 416-480-6100 ext. 63356 or pstefanova@sri.utoronto.ca.
Helpful links
The histology core facility offers paraffin services including the tissue processing, embedding, sectioning, and various staining for tissues. One-on-one services are available to discuss tissue processing, embedding orientation and staining preferences. Additionally, the facility provides frozen and vibratome services.
The facility offers training and self-access to three pieces of equipment – microtome, cryostat and vibratome.
- Tissue processing: Tissue processing starts with a fixation step that preserves the tissue followed by steps that dehydrate, clear and infiltrate the tissue with paraffin wax.
- Decalcification: This technique removes mineral from bone and other calcified tissues so that good-quality paraffin sections can be prepared.
- Paraffin embedding: This process creates paraffin blocks from specimens that have been thoroughly infiltrated with paraffin wax. The blocks can be clamped into a microtome for sectioning.
- Sectioning: Using a microtome, very thin sections are cut and placed on a microscope slide for staining.
- Routine staining: Routine staining uses hematoxylin and eosin dyes to stain thin tissue sections, which allows tissue morphology to be visualized.
- Special staining: Special stains are useful for identifying specific cell types, demonstrating certain cellular components, or highlighting particular pathological features.
- Cryosectioning: Cryosectioning is an alternative tissue preparation technique that quickly freezes tissue to preserve it. This technique is generally used when chemical fixatives need to be avoided.
- Vibratoming: This procedure uses a vibrating blade microtome to cut sections of fixed or fresh tissue under physiological conditions without freezing or embedding.
For self-service access to the SRI histology facility, principal investigators (PIs) are required to do the following:
- Fill out and submit an online SRI histology facility PI membership form.
- Pay an annual membership fee for trainees to have access to the facility ($600 for SRI members; $1,200 for non-SRI members).
- Pay an hourly user fee ($30 per hour for SRI members; $45 per hour for non-SRI members) or annual user fee ($2,000 per year for SRI members; $4,000 per year for non-SRI members). Note: This fee applies to all pieces of equipment. For self-service access to the SRI histology facility, trainees are required to do the following:
- Complete and submit an online SRI histology facility orientation form.
- Read, complete and sign the SRI histology facility service agreement form (.docx). Note: This form requires your PI’s signature. Bring the completed form to the histology facility.
- Complete and submit an online SRI histology facility training form. Sign up for a training session on the equipment to be used. Times to be arranged with the facility manager, Petia Stefanova.
- For trainees paying hourly fees, fill out the online SRI histology facility self-service form and submit it. Note: This form is not required for annual users.
Additional details on the terms and conditions of use are provided in the SRI histology facility service agreement form (docx).
Self-service fee schedule
The following fee schedule is for open access to the facility for SRI members.
| Description | Fees |
|---|---|
| Access fee is an annual fee for PIs who wish to have trainee access | $600 |
| Histology annual usage fees (capped to ensure affordability for our members) | $2,000 |
| No-show or cancellation fee (without 24-hour notice) | $50 |
| Facility orientation fee | $50 per trainee |
The following fee schedule is for open access to the facility for non-SRI members.
| Description | Fees |
|---|---|
| Access fee is an annual fee for PIs who wish to have trainee access | $1,200 |
| Histology annual usage fees (capped to ensure affordability for our members) | $4,000 |
| No-show or cancellation fee (without 24-hour notice) | $75 |
| Facility orientation fee | $75 per trainee |
We offer training on the following equipment that can then be used by trainees independently.
| Equipment | Training Fee | Refresher Fee | Usage Fee | |||
|---|---|---|---|---|---|---|
| SRI | non-SRI | SRI | non-SRI | SRI | non-SRI | |
| Leica CM3050 S cryostat* Must provide your own anti-roll plate, disposable blade and microscope slides, slide boxes or racks | $125/hr | $187.50/hr | $62.50/hr | $93.75/hr | $30/hr | $45/hr |
| Leica RM 2255 automatic rotary microtome Must provide your own disposable blades, microscope slides, slide boxes or racks | $125/hr | $187.50/hr | $62.50/hr | $93.75/hr | $30/hr | $45/hr |
| Leica VT1000 S vibrating blade microtome (for histology) Must provide your own disposable blades, PBS for sectioning, plates to collect sections in | $125/hr | $187.50/hr | $62.50/hr | $93.75/hr | $30/hr | $45/hr |
| Leica VT1000 S vibrating blade microtome (for live slice culture) Provide reagents as above. Also arrange in advance to have chamber and sample holder autoclaved | $125/hr | $187.50/hr | $62.50/hr | $93.75/hr | $30/hr | $45/hr |