Department head
Dr. Sabine Hombach-Klonisch
sabine.hombach-klonisch@umanitoba.ca
The University of Manitoba campuses are located on original lands of Anishinaabeg, Cree, Oji-Cree, Dakota, and Dene peoples, and on the homeland of the Métis Nation. More
University of Manitoba
Winnipeg, Manitoba Canada, R3T 2N2
The histology laboratory provides services for researchers and students to process their tissues and cells for histopathological examination.We offer paraffin, cryo, and electron microscopy techniques including processing, embedding, sectioning, staining and imaging. We have an array of common and special histological stains available for specific projects. Selected immunofluorescent and immunohistochemical detection and special assays are also available upon request. Histomorphometry studies related to changes in histopathology of samples can be identified using new techniques in imaging and analysis.
To request histology services, electron microscopy or imaging facility services, please contact the histology lab for a consultation.
After the consultation, we will provide an estimate for your approval. We will then let you know how to prepare your samples for optimal results and schedule an appointment for sample drop off. Please do not bring samples to the lab without an appointment.
Samples must be accompanied by completed forms (work request form, sample submission form, sample waiver form) as well as the signed estimate.
We offer standard paraffination preparation.
To set up a consultation to discuss your specific needs or ask any additional questions, please contact the histology lab.
Carefully read the guidelines and ensure you are bringing properly prepared samples.
Samples must be grossed to a reasonable size BEFORE fixation.
Tissues should be no thicker than 5mm and a maximum of 1.5cm length and width.
Smaller pieces have better fixation and are easier to section.
Provide only tissue significant to your research, having grossed out any other attached tissue or organs that are not necessary.
If you deal with larger specimens, please provide only a biopsy.
Please fix all tissues in appropriate fixative based on what the samples are to be used for.
Perfusion is optimal if using animal tissues, immersion also sufficient if the tissue is of an appropriate size.
Some fixatives may interfere with staining or immunohistochemistry conditions.
Common fixatives used are 10% buffered formalin or 4% formaldehyde (prepared from a 37% stock liquid containing methanol commercially purchased or prepared from a powder).
Minimum fixation time should be 24 hours and can last as long as five days.
Your tissue sample should be free floating in fixative of at least 10X the tissue volume.
Once your samples are in fixative, they may be delivered to the histology lab. Samples may also be stored in PBS at 4°C or 70% ethanol at room temperature for short periods.
In addition to your samples, please bring:
Important: Please make arrangements with histology staff PRIOR to sample drop off to ensure we are available to accept your samples.
The pricing shown here is for UM and affiliated investigators. For industry pricing, please contact us.
Task | Description | Item | Cost |
---|---|---|---|
Processing tissue for paraffin | Placing tissue that arrives in fixative (already grossed) into cassettes and labeling for the tissue processor | Fixative, formalin, dehydrating alcohol, xylene and paraffin | included in processing charge |
1-24 cassettes | $39 | ||
25-50 cassettes | $78 | ||
51-75 cassettes | $117 | ||
76-100 cassettes | $156 | ||
Labour | $40/hour | ||
Embedding tissue in paraffin | Taking the cassettes from the tissue processor and setting them in paraffin in the correct orientation, then filling the mold with paraffin to create the block | Paraffin from embedding machine | Included in tissue processing charge |
Paraffin, if separate | $5 each | ||
Cutting paraffin sections | Sectioning the desired number for sections/slide from each block as requested | APTES | $1/slide |
Superfrost plus | $1.50/slide | ||
Knife blade | included | ||
Labour | $40/hour | ||
Staining paraffin sections | Staining and cover slipping slides as requested | Varies depending on the requested stain | See staining information for details |
H&E | $3/slide | ||
Labour | $40/hour | ||
IHC, IF and other assays | IHC or IF on the prepared slides with antibodies as well as other requested assays (kit) | Custom pricing for individual requests | $100/run plus all required supplies and chemicals |
Labour | $40/hour | ||
Imaging/analysis | Image the slides using bright field or fluorescent microscopetaking pictures, performing analysis where required (counting, measuring, graphing) | UM | $75/hour |
External, imaging done by staff | $100 | ||
USB | Saving image to USB for client | USB | $8 |
Digital transfer | Digital image transfer via online platform | N/A | $0 |
Slide box | Slide storage/transport | Box | $10 |
We offer standard frozen preparation.
To set up a consultation to discuss your specific needs or ask any additional questions, please contact the histology lab.
For best results, tissues must be properly prepared.
Carefully read the guidelines and ensure you are bringing properly prepared tissues and all required paperwork and approvals.
Samples must be grossed to a reasonable size BEFORE fixation or snap freezing.
Smaller pieces have better fixation and are easier to section. Tissues should be no thicker than 5mm and a maximum of 1.5cm length and width.
Provide only tissue significant to your research, having grossed out any other attached tissue or organs that are not necessary. If you deal with larger specimens, please provide only a biopsy.
Please fix all tissues in appropriate fixative based on what the samples are to be used for. Some fixatives may interfere with staining or immunohistochemistry conditions.
Common fixatives used are 10% buffered formalin or 4% formaldehyde (prepared from a 37% stock liquid containing methanol commercially purchased or prepared from a powder).
After 24 hours fixation, wash the tissue in 1 x PBS, three times for five minutes.
Make a 20% sucrose solution in 1 x PBS and transfer the tissue into the solution. Keep the tissue in this solution at 4°C until it sinks (usually overnight).
Make a 30% sucrose solution in 1 x PBS and then transfer the tissue from 20% sucrose solution to the 30% sucrose solution at 4°C again until the tissue sinks (usually overnight).
Transfer the tissue from the 30% sucrose solution to pure OCT or desired embedding medium) for up to 24 hours at 4°C.
Freezing artifacts are common, please ensure you are freezing the blocks correctly (quickly enough) to avoid this. Freezing blocks too slowly results in poor morphology.
If tissues are to be unfixed, they may be snap frozen in isopentane/liquid nitrogen in OCT and delivered frozen to the histology lab.
Please ensure blocks are correctly frozen and not cracked and tissue orientation is correct. Incorrect preparation will result in freeze artifacts.
Once your samples are in fixative, they may be delivered to the histology lab. We will also accept already blocked samples frozen in cryo molds with OCT or other desired medium.
In addition to your samples, please bring:
Important: Please make arrangements with histology staff PRIOR to sample drop off to ensure we are available to accept your samples.
The pricing shown here is for UM and affiliated investigators. For industry pricing, please contact us.
Task | Description | Item | Cost |
---|---|---|---|
Fixing/embedding tissue for cryo sectioning | Fix tissues in 4% paraformaldehyde, infuse with 20% and 30% sucrose, infiltrate with OCT and embed tissues in OCT in molds | Fixative, PBS, sucrose, OCT, molds, dry ice | $9 each |
Labour | $40/hour | ||
Embedding tissue for cryo sectioning | Fixed or snap frozen tissue is placed in molds with OCT and frozen | OCT, molds, dry ice | $5 each |
Labour | $40/hour | ||
Cryo sectioning | Sectioning the desired number for sections/slide from each block as requested | APTES | $1/slide |
Superfrost plus | $1.50/slide | ||
knife blade | included | ||
Labour | $40/hour | ||
Staining frozen sections | Staining and cover slipping slides as requested | H&E | $3/slide |
Varies | See staining page for special stains | ||
Labour | $40/hour | ||
IHC, IF and other assays | IHC or IF on the prepared slides with antibodies as well as other requested assays (kit) | $100/run | |
All required supplies and chemicals | varies | ||
Labour | $40/hour | ||
Imaging/analysis | Image the slides using bright field or fluorescent microscopetaking pictures, performing analysis where required (counting, measuring, graphing) | UM | $75/hour |
External, imaging done by staff | $100 | ||
USB | Saving image to USB for client | USB | $8 |
Digital transfer | Digital image transfer via online platform | N/A | $0 |
Slide box | Slide storage/transport | Box | $10 |
Using a variety of staining processes, we can provide visual details about protein abundance, distribution, and localization.
To set up a consultation to discuss your specific needs or ask any additional questions, please contact the histology lab.
We provide TEM services for tissues, cells and nano-particles using the Phillips CM10.
Tissue and cell samples are typically embedded in Embed 812 and sectioned with a diamond. Thick sections are 0.5-1um and stained with toluidine blue for orientation, quality control and documentation. Thin sections are 90-100nm collected on grids. Staining is done using uranyl acetate or uranyless and lead citrate. Imaging is done at 800 - 64,000X magnification.
To set up a consultation to discuss your specific needs or ask any additional questions, please contact the histology lab.
For best results, tissues must be properly prepared.
Carefully read the guidelines and ensure you are bringing properly prepared tissues and all required paperwork and approvals.
Tissues must be grossed to 1mm cubes before fixation.
Provide only tissue significant to your research, having grossed out any other attached tissue or organs that are not necessary. If you deal with larger specimens, please provide only a biopsy.
Please fix all tissue or cells in 3% glutaraldehyde in 0.1 M Sorensen’s buffer.
This can be provided by the histology lab, please make arrangements for fixative pick-up in advance of cell harvest or animal sacrifice so we can ensure the solutions are fresh.
If processing brain or nerve tissue, a combination of parafomaldehyde and gluteraldehyde may be provided.
If using animals, perfusion fixation is imperative for preservation of fine structures.
Two to three hours for cells and three to four hours for tissue.
Your tissue sample should be free floating in fixative of at least 10x the volume of the tissue.
Once your samples are in fixative or sucrose solution (also may be provided) they may be delivered to the histology lab at a pre-arranged appointment time with the completed paperwork:
A slide box is not required for TEM samples.
The pricing shown here is for UM and affiliated investigators. For industry pricing, please contact us.
Task | Description | Item | Cost |
---|---|---|---|
Processing and embedding tissue for EM |
Fixing tissue or cells that arrive (already grossed) in fixative, sucrose buffer through to embedding in plastic |
Fixative, sucrose, buffer, osmium tetroxide, alcohol dehydration, propylene oxide, EmBed 812 |
Included |
1-10 blocks |
$195 |
||
Labour, five hours per tissue set and six hours per cell set |
$40/hour |
||
Sectioning (diamond knife) |
Thick sectioning and staining with toluidine blue, thin sectioning 90-100nm and placing on desired number of grids (200 or 300 mesh copper) nickel or carbon/formvar available upon special request |
diamond knife, slides, toluidine blue, grids |
Included |
Labour |
$50/hour |
||
Staining grids |
Uranyl acetate, lead citrate or uranyless, lead citrate |
Included |
|
Labour |
$50/hour |
||
Imaging Electron Microscope Phillips CM10 |
TEM imaging done by histology staff |
UM faculty and affiliates |
$75/hour |
External |
$120/hour |
||
USB | Saving image to USB for client | USB | $8 |
Digital transfer | Digital image transfer via online platform | N/A | $0 |
Superresolution structured illumination microscopy (SR-SIM) is an advanced imaging technique used to image fine structural details with minimal changes to standard immunofluorescence protocols using conventional dyes.
The resulting image is of high quality with up to 120 nm lateral resolution (X,Y) and 300 nm axial resolution (Z).
This technique uses five different grating frequencies to allow for optimal matching of illumination pattern to laser wavelength and objective lens used.
For more information about SR-SIM, contact Dr. Thomas Klonisch at 204-789-3893.
Shown here are fluorescently labelled cells imaged in widefield and with structured illumination processing comparison.
To request training and/or access, complete the request form.
Use the links below for quick access to our forms.
Dr. Sabine Hombach-Klonisch
sabine.hombach-klonisch@umanitoba.ca
Dr. Thomas Klonisch
thomas.klonisch@umanitoba.ca
Dana Henderson
dana.henderson@umanitoba.ca
Farhana Begum
farhana.begum@umanitoba.ca
Histology Services, Imaging Facility, Electron Microscopy Platform
Room 119, 745 Bannatyne Avenue
Basic Medical Science Building
University of Manitoba (Bannatyne campus)
Winnipeg, MB R3E 0J9