MICROSCOPIE

Plateforme d'imagerie et microscopie de l'IBIS

User Tools

Site Tools


en:demande_de_service:explications

How to Submit

Sample Preparation Request

Coordinates

Your contact details are necessary to identify you. We cannot process a sample submitted with incomplete contact information. To obtain the ULaval rate, you must provide a project number (numbers beginning with CG, FT, IC, etc.). Otherwise, for NPOs, public and parapublic organizations and businesses, rates are available via the website.

Samples

Number: If your samples can be pooled for embedding in the same cassette, the number of samples will match the number of cassettes.

What solution is the specimen in?: Indicate the solution, buffer, fixative, solvent, etc. If the solution presents a danger other than those related to fixers or solvents, indicate this.

Fixation: Fixation is a critical step in microscopy and fixatives are not the same in electron microscopy as in histology. Refer us to our website for more information. If your specimen needs to be fixed by the microscopy platform, please contact us for an appointment. You can get Fixer Ampoules from the platform as needed.

Storage: Especially if your sample is not fixed, be sure to include an expiration date and storage temperature.

Identification: Don't forget to complete the sample identification form. You can enter particularities or different treatment per sample.

Optical Histology

Work to be done: Indicate only the steps to be done.

Cuts per slide: It is possible to put 1 or 2 cuts per slide, depending on the size of the specimen.

coverslips: #1.5 coverslips are the desired standard for high resolution microscopy (for oil objectives). Slides #1 are thinner and are sometimes appreciated for easier reading in some automatic imagers.

Number of slides per sample and choice of stains: The number of slides desired per sample. Write “max” if it is a depletion of the specimen. In addition to standard Hematoxylin & Eosin staining, a variety of stains are available on our website.

Notes and Parameters: Mention in the bottom section any important information. Does the orientation of the specimen matter? Should we make serial sections, that is to say successive sections without failure in order to image a volume? Should we cut the whole specimen or on the contrary be sure to keep some for the future? Is there a particular cutting thickness?

Electron Microscopy

Electron microscopy comes in three parts. Ultramicrotomy for transmission microscopy (TEM), direct deposition for TEM and preparations for scanning microscopy (SEM). Check the appropriate box(es).

TEM: Ultramicrotomy

Embedding: It is possible to embed the sample in an epoxy (EPON) or acrylic (LR-White) matrix. Typically EPON is more routinely used. However, LR-White has the advantage of allowing immuno-detection (immuno-gold) and dehydration remains partial. If you want to do immunodetection, the fixation must be appropriate (see our website). For embedding, the specimens are cut into pieces smaller than 1 mm. The embedding package automatically gives 5 blocks per sample, if your specimen allows it.

Cuts: Semi-thin cuts are included in the embedding package. They are typically stained with toluidine blue. Ultra-thin cuts are extra. These 80 nm sections are typically mounted on grids. Normally, 5 grids per sample are provided, including 2 colored (3 kept for adjustments). Color adjustments are free of charge.

Staining: Staining is done with uranyl acetate and/or lead citrate, but other options are possible. Visit our website.

TEM: Direct Deposit

With this technique, the solution is deposited directly on a grid with a carbon film. The excess is absorbed with a filter paper so as to create a thin film. For samples of solid nanoparticles, or virus particles, this technique works quite well. For food matrices or polymers, some experimentation is required to avoid drying artifacts. Enter the solvent and the desired dilution. If the particles form aggregates, it is possible to use ultrasound before dispersion (mention in the notes section). Enter the desired coloring, if necessary.

SEM

Drying: Generally, samples should be dried before visualization except when using low vacuum SEM. Dehydration is done by alcohol gradient and finished with hexamethyldisilazane (HMDS) or liquid CO2 at the critical point.

Metallization: Except for conductive samples, it is necessary to metallize the sample with a gold layer deposit.

Website

en/demande_de_service/explications.txt · Last modified: 2022/02/02 15:51 by Administrateur

Donate Powered by PHP Valid HTML5 Valid CSS Driven by DokuWiki