© Histology Services, Imaging Facility & Electron Microscopy Platform, University of Manitoba, Canada,  2022

Cryo

We offer standard frozen preparation

To set up a consultation to discuss your specific needs or ask any additonal questions please contact the histology lab. Please make arrangements with histology staff PRIOR to sample drop off so we can be available to accept your samples. Carefully read the guidlines below and ensure you are bringing properly prepared samples, and all required paperwork and approvals.

Cryo Sample Preparation Guidelines:

For best reults tissues must be properly prepared. Size - Samples must be grossed to a reasonable size BEFORE fixation or snap freezing. 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. Fixation - 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, 3 times for 5 minutes. - Sucrose infuse tissue (cryoprotection): 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. - Infiltration: 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 at a pre-arranged appointment time with the completed Work Request, Sample Submission and Waiver Forms. We will also accept already blocked samples frozen in cryo molds with OCT or other desired medium. Please bring a slide box of sufficient size for the requested slides at the time of sample drop off or one will be added to your invoice.

Task

Description

Chemicals Cost For Service

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
$9.00 each $40.00/hour labour
Fixing/Embedding tissue for cryosectioning
Embedding Tissue for cryosectioning
Fixed or snap frozen tissue is placed in molds with OCT and frozen
OCT, molds, dry ice
Cryosectioning
Sectioning the desired number for sections/slide from each block as requested
knife blade included
$40.00/hour labour $1.00/slide APTES $1.50/slide Superfrost Plus
Staining frozen sections
Staining and coverslipping slides as requested
Varies depending on the requested stain
$40.00/hour labour $3.00/slide H & E SeeStaining page for special stains
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
$100.00/ run $40.00/hour labour plus all required supplies and chemicals
Imaging/analysis
Image the slides using bright field or fluorescent microscope taking pictures, performing analysis where required (counting, measuring, graphing)
Digital Transfer USB
Digital image transfer via online platorm Saving image to USB for client
cost of USB
no charge $8.00/USB
$5.00 each $40.00/hour labour
Slide Box
Slide storage/transport
cost of box
$10.00/Box

Cryo Service Pricing

HISTOLOGY SERVICES IMAGING FACILITY ELECTRON MICROSCOPY PLATFORM
Please contact the histology lab for customized industry pricing
$75.00/hour U of M $100.00/hour External Imaging Done by Staff