Reception and Delivery Hours
10-11 - 13-14 h

Equipment Reservations

• Equipment reservations (cryostats) must be formally made through Intranet-Reservations-Equipment. Established rules regarding equipment use and general operation must be followed.
• A maximum of 2 hours can be reserved by the same user and laboratory, with a maximum of 8 hours per week. If reserved hours are not to be used, they must be canceled at least 1 hour in advance. In case of cancellation with less than one hour notice, the Facility must still be informed. If 30 minutes pass from the start of the reservation without notifying the reasons for the delay, the Facility will proceed to cancel the reservation and call the users on the waiting list.
• If more hours are required due to sample characteristics, contact the Facility for authorization or redistribution based on demand.
• The cryostat 1950 located in the Instrumentation Room (2nd floor IIB2) may also be used by authorized users under the supervision of the Facility staff. Authorized users are those recognized by the supervisor as having sufficient experience with the equipment.
• Any doubts, problems, or malfunctions of the equipment should be reported as soon as possible to the service supervisor through the incident page (Cyberlar-Incidents-Histology) or by phone.

Sample Submission Guidelines

• The user must request the service via intranet/purchases/histology before sample submission. In Notes, indicate: orientation of organs, if a preliminary cut is needed, area to be cut, number of slides/samples, if multiple different samples are to be placed on each slide, or any specific requirements.
• For external account billing, the user must complete the corresponding request and submit it to the service along with the samples. Both must fill in the sample reception and preparation registry for histology.
• Samples must be clearly labeled with the principal investigator's name (initials) or laboratory and numbered with a specific code (consecutive numbers). Label the cassettes with a pencil. Send an excel or list to the service with the submitted samples.
• For frozen human samples,indicate if the sample has been previously fixed.

Sample Collection Guidelines

• Histology staff will contact the designated person once all requested services are completed and reviewed to inform them to collect the preparations. Samples will be on the Service's entry table labeled with the researcher's/user's name and order number. Check and sign the sample collection registry.
• Samples for cryostat cutting can be left in ultrafreezer no. 2, located in the basement in P. Animalario, in the drawer designated for the Histology Service (shelf 5) and the B-39 tray. The boxes with samples must be perfectly identified with the user and the Histology Facility's name. Notify the delivery and collection of material and sign the corresponding notebooks

1. Paraffin Block Preparation

• Samples must be fixed in 4% paraformaldehyde or formaldehyde (according to the researcher’s criteria) with a volume 20 times greater than the sample. The sample size should be approximately 1 cm³.
• Samples should remain in the fixative overnight at 4°C or for 1-2 hours at room temperature. The fixation time depends on the sample’s source and size.
• If it takes more than 24 hours to deliver the sample, wash it in PBS 1x for 30 minutes (2-3 times) and store it at 4°C until delivery.
• Note in the logbook how long the sample has been in the fixative or whether it is delivered in PBS1x or 70% ethanol. It is recommended to deliver the sample within the embedding cassette (request in advance), already cut and cleaned. If a cut is required before processing, indicate the plane and whether decalcification is needed.

2. OCT Block Preparation

• OCT blocks can be prepared with or without fixation.
• For OCT block preparation by the service, samples are fixed in 4% paraformaldehyde or formaldehyde and then processed in 30% sucrose (in PBS 1x) for at least overnight (O/N). They can be delivered in either the fixative or the sucrose.

3. Paraffin and Frozen Sections

• Indicate the number of slides required for each sample and whether a specific number of sections per slide is needed. If not specified, the sections will be made at 5 microns.
• Billing is done in intervals of 10 cut slides or 5 cut slides. Billing is per slide, not per section.
• Provide the necessary boxes for the slides with the sections.
• For floating sections, indicate how many series are needed per sample. Provide Eppendorf tubes or plates and the antifreeze solution. Billing is per sample.
• Paraffin, OCT, or gelatin blocks can also be delivered directly for further processing.

4. Staining

• In addition to the stains offered on the website (HE/E, Oil Red O, HE Nissl), specific stains can be provided, and the service will perform the staining by requesting "Other Stains for Paraffin Sections."

Rates:

• Tarifas histología 2023 ( a partir del 1 de marzo 2023)
• tarifas histología 2024 (a partir del 1 de marzo 2024)
• tarifas histología 2024 actualización UAM

Cryostat 1950 Usage Rules

A. General Access and Reservation Rules

• Only authorized personnel with prior experience in this equipment can use it. Any problems caused by improper use will be the responsibility of the user and, ultimately, the principal investigator. These users must know and follow the operating, cleaning, and safety rules established by the Facility. They must also be registered in the autonomous cryostat user log.
• Users without prior experience must contact the  manager Facility for proper training.
• Only 2 hours per user/laboratory and 8 hours per week can be reserved. If an hour becomes available, it can be reserved with the permission of the Facility staff. This reservation may be canceled if requested by a user without any reserved hours.
• Access to the Instrumentation Room, where the cryostats are located, is controlled. To gain access, request the key at reception and fill out the corresponding logbook (user, laboratory, entry and exit times, incidents). The user is responsible for locking the room and returning the key to reception after use.
• Each user must bring their cleaning materials and accessories (ethanol, brushes, blades, OCT, slides).
• Reservations are mandatory. If the reservation will not be used, it must be canceled at least 1 hour in advance, and all users must be notified via email. Reservations are made via intranet. If there is no reservation, notify Histology to be added to a waiting list, and the Facility will call when a reservation is canceled.

B. General Cryostat Usage Rules

• LOCK the handle whenever a sample is placed or moved. After finishing work, lock it again.
• Before starting to cut, remember to unlock the handle and avoid forcing it with the brake engaged.
• DO NOT TOUCH the snow button when defrosting.
• DO NOT CHANGE the angle of the knife holder. It is recommended to keep it between positions 3-5.
• Ensure that the anti-roll plate is well-positioned to avoid friction during cutting (the anti-roll plate sticks out too far from the knife edge), as this may damage the plate. Avoid scratching the plate.
• To avoid vibrations that may affect the cut thickness:
  • Tighten the handle well to orient the sample.
  • Place enough OCT on the sample holder before positioning the sample. Perform this process at room temperature. Once the sample is placed, insert it into the cryostat chamber.
  • Make sure the two holding levers for the knife holder and the lever for adjusting the blade are tight.
  • The knife holder should not be too far out (at least 1 cm from the outer edge). When starting work, move the knife holder as close as possible to the sample.
  • If the arm is too far out while cutting, push it back and move the knife holder closer to the sample.
• After finishing work:
  • Remove the blade.
  • If the anti-roll plate was removed, put it back in place.
  • Leave the arm fully retracted and locked.
  • Clean any residue with 70% ETHANOL (never with water) from the trays or inside the chamber.
  • Ensure that all levers are tightened.
  • Return the sample holder discs to their place and clean any residue with ethanol.
  • Close the cryostat glass door.
  • If the temperature was changed, reset it to the original setting.
  • Turn off the chamber light.
  • Fill out the cryostat logbook with the entry and exit times.
  • Disconnect the sample holder cooling system.

NOTE: If the "Key" button is held for more than 5 seconds, the equipment will lock. The sign that it is locked is that the ":" symbol between the hour/minute on the clock panel disappears. To unlock, hold the "Key" button for 5 seconds until the ":" symbol reappears.

• Do not turn off the chamber cooling system.
• Do not leave any materials on the countertops.
• Never turn off or adjust the air conditioning temperature.
• Lock the door and return the key to reception.

The last registered user is ultimately responsible for any material loss or misuse of the equipment. For example, there should always be two sample holder discs. In case of any issues with the equipment, contact the Manager Facility, Yolanda Ortega (yortega@iib.uam.es or the Histology Office phone extension 443865).

C. Specific Rules for Cryostat 1950

• Only users authorized by Histology can use it. To use it in the afternoon, prior supervision during Facility hours is required.