Below you will find the terms and conditions of use of the UMIB Core Facility to make your experience with us as pleasant as possible.
Through the UMIB Core Facility you will have the opportunity to access state-of-the-art equipment that will assist you in the development of your scientific research, that is, a Seahorse Extracellular Flow Analyzer, a Stopped-Flow Analyzer and a Thermocycler.
Location
UMIB Core Facility is located at ICBAS-UP, at Rua Jorge Viterbo Ferreira, no. 228, 4050-313 Porto, Building 1, Piso 5, Sala 3.
- The provision of services is carried out by the Researcher Responsible for the UMIB Core Facility or the Co-Responsible Investigator.
- UMIB Core Facility’s laboratory opening hours are from 9:00 a.m. to 6:00 p.m. on weekdays.
- The provision of services is subject to prior reservation and is restricted to:
a) Researchers integrated into UMIB.
b) Other researchers from academic or business entities. - Users must, prior to booking the equipment (at least 5 working days), request their quote by email. If you wish to make the reservation request, you must register using the following email.
- The provision of the service will be preceded by a meeting with the Researcher or Co-Investigator in charge of the UMIB Core Facility to define appropriate procedures and protocols to ensure the quality of the results obtained in the provision of services. The procedures set out at this meeting shall be recorded in a document signed by the interveners.
- On the date set for the provision of the service, the user must attend the UMIB Core Facility 15 minutes before the scheduled time, accompanied by the samples.
- During the performance of the provision of services should remain with the respective equipment, and it is up to it to monitor the analysis of the samples under study, their conservation and storage.
- Users may not handle any material or equipment without prior authorization from the Researcher or co-investigator in charge of the UMIB Core Facility.
- Users must comply with the “General Laboratory Safety Rules” and the instructions given by the Researcher or co-investigator in charge of the UMIB Core Facility.
- Failure by the user to comply with the conditions defined in these conditions of use and causing damage to the goods, equipment and/or the premises, resulting from the user’s deed or negligence, shall be imputed to the user, and shall bear financially the damage caused, and, where appropriate, the corresponding legal damages that may be determined.
Equipment
Extracellular flow analyzer
Seahorse XFe24 Analyzer
The Seahorse XFe24 Analyzer uses 24-well plates with a maximum injection capacity of 4 distinct compounds.
Expected results
The Seahorse XFe24 Analyzer evaluate the oxygen consumption rate (OCR) and the extracellular acidification rate (ECAR) of living cells, tissues, and mitochondria. OCR and ECAR rates are key indicators of mitochondrial respiration and glycolysis, as well as ATP production rate, and together these measurements provide a system-level view of cellular metabolic function in cultured cells and ex-vivo samples. The use of this equipment allows the use of smaller equipment (centrifuge, greenhouse and microscope).
Notes
- The consumables used in the Seahorse XFe24 Analyzer have to be purchased from Agilent because it is the only company that markets materials compatible with the equipment. https://www.agilent.com/search/?Ntt=XFe24
- The consumables used in the Seahorse XFe24 Analyzer have to be purchased from Agilent because it is the only company that markets materials compatible with the equipment. https://www.agilent.com/search/?Ntt=XFe24
Preparation of samples The method of preparing samples for reading in the Seahorse XFe24 Analyzer will depend on the type of sample: Suspended cells and cell organelles
- Select the desired cell density or cell organelles (can be tested previously on a plate of 96 well, the well should have about 80% confluence)
- Select the best coating for the plate to the cells or organelles under study (e.g., Polylysine, Collagen, etc.).
- Pre-coat the reading plate of the Seahorse XFe24 Analyzer, following the recommendations of the compound used for the coating.
- Transport the previously coated plate and the cells or organelles to the UMIB Core Facility laboratory.
- Dilute the cell pellet or organelles in the Seahorse reading medium, and seed the cells at the desired concentration.
Adherent cells
- Select the density of cells so that at the time of the assay the cells reach about 80% of confluence (can be tested previously on a plate of 96 well).
- Seed the cells on the Reading Plate of the Seahorse XFe24 Analyzer;.
- Cells must reach the confluence desired by the user.
- The culture medium should be replaced by the Seahorse reading medium and the plate should be incubated during 1 hour in a CO2 free incubator.
- The Seahorse reading medium must be replaced by fresh heated seahorse medium and the plate is ready to be placed in the Seahorse XFe24 Analyzer.
Tissues
- Use the tissue as fresh as possible.
- Cut the tissue into small fragments (2-5mg) and gently wash each fragment in Seahorse reading medium (heated to 37ºC).
- Place each tissue fragment in the center of each well of the Seahorse XFe24 Analyzer reading plate.
Equipment
Stopped-Flow Analyser
SX20-LED
The SX20-LED Stopped-flow Spectrometers is a high-resolution spectrometer with high-intensity LED light source with available wavelength at 415 nm. It requires low sample volume and acquires results with low “dead time”.
Mix in proportion two solutions, simultaneously triggering both syringes by a single plunger for continuous, fast, and efficient mixing in 1:1 proportion.
Expected results
- Used to study rapid reaction mechanisms (1ms – 100 seconds).
- Two reagents are quickly mixed, still in an observation cell and spectroscopic properties are measured as a function of time.
- Absorbance is measured at 415 nm.
The analysis of spectroscopic signals can determine reaction rates, intermediateshort-lived, give an insight into the mechanisms of molecular processes, … - A series of interrupted flow experiments can be used to show the effect of temperature, pH, and reagent concentration on reaction kinetics.
Notes
- The modulators/consumables to be used must be ensured by the user.
- The use of this equipment allows the use of smaller equipment (centrifuge, incubator without CO2, microscope and osmometer).
Preparation of samples The method of preparing samples for reading on the SX20 Stopped Flow Spectrometer will depend on the sample type:
Organelles/cell vesicles
Organelles (e.g., mitochondria) or cellular vesicles (obtained through cell membranes) can be isolated from cells or tissues. Cellular organelles or vesicles should be isolated prior to the experiment, with user-defined protocols. The biological material must be transported to the UMIB Core Facility laboratory in an isotonic medium (e.g., PBS).
Intact cells
- Suspended cells: Centrifuge the cell suspension and wash the cell pellet with PBS. Dilute the cell pellet in an isotonic solution.
- Adherent cells: Tripinize cells and ensure separation of cell aggregates. Wash the cell pellet with PBS. Dilute the cell pellet in an isotonic solution.
Equipment
Thermocycler
T100 Thermal Cycler
The T100 thermocycler is a compact thermocycler that has a user interface with intuitive touch screen to facilitate PCR execution. Thermal gradient technology allows you to quickly optimize your reaction in a single run.
Expected results
- Nucleic acid amplification (PCR)
- Cloning and gene analysis
- Gene expression analysis
- Mutational analysis
Preparation of samples
The method of preparing samples for the Thermocycler is the responsibility of the user, so the protocols used for the extraction of nucleic acids (DNA or RNA) must be prepared prior to the use of the equipment.
- The preparation of the samples until the time of their analysis is the responsibility of the user, committing the user to fully comply with all the rules defined for the provision of service.
- It is up to the user to monitor the analysis of the samples under study, to ensure their storage and, if applicable, their storage.
- Exceptionally, and since previously established, UMIB core facility may ensure the temporary preservation of samples for the agreed period, after which, if not collected, they will be discarded.
The price* to pay for each service varies depending on the type of user associated with (does not include the necessary consumables).
In cases expressly provided for and identified in the budgeting, the price may be increased by the cost of consumables used in the preparation/storage of samples.
| Users | Seahorse XFe24 Analyzer | Stopped Flow Spectrometer SX20 | Thermocycler T100 |
|---|
| User type | Price / hour | ||
| C1 | 25€ | 10€ | 10€ |
| C2 | 40€ | 15€ | 15€ |
| C3 | 50€ | 25€ | 25€ |
| User type | Description |
|---|---|
| C1 | Integrated researchers from UMIB or the Clinical Academic Center of the Abel Salazar Institute of Biomedical Sciences and The University Hospital Center of Porto (CAC ICBAS-CHUP). |
| C2 | Researchers integrated in organic units of the University of Porto. |
| C3 | Researchers from other academic and research institutions, of a public or private nature. |
- Consumables used in The Seahorse XFe24 Analyzer equipment can be purchased directly by the user or may be provided by UMIB Core Facility upon acceptance of a quote request for this purpose.
- The modulators and consumables to be used on Stopped Flow Spectrometer SX20 and Thermocycler T100 equipment, respectively, must be insured by the user and may not be supplied by UMIB Core Facilty.
* The amounts shown do not include VAT.
Cancellation of the reservation must be made by email, at least two working days before the booked date, and the amount is due.
- UMIB Core Facility ensures that the results obtained to users are downloaded by e-mail within 24 hours of the service being serviced.
- The results of the service provision will be sent without treatment of the information contained therein, and it is exclusively up to the user to analyze and interpret the primary experimental results
obtained. - With the transfer of the results to the user ceases the responsibility of the UMIB Core Facility.
- All communications to be made by UMIB Core Facility will be formalized through the email address.
- In case of urgency, the telephone contact may be used for the number indicated on this page, without prejudice to the subsequent sending of an e-mail.
With the delivery of the result of the provision of services, there is the transfer of ownership and ownership of the elements produced, to the user.
Specific services may be agreed for situations duly analysed and framed by the UMIB Core Facility and which require a more elaborate and/or prolonged preparation of the samples to be tested, in the terms and conditions that may be defined.
UMIB Core Facility will be responsible for/to:
- Comply with and enforce the guidelines contained in this regulation.
- Ensure the proper functioning of the UMIB Core Facility and the correct use of the equipment.
- Ensure, in due course, that the conditions for the performance of each service are met.
- Supervise the careful and rational use of UMIB core facility materials and equipment.
- Manage the appointments and reservations, providing all necessary clarifications to users.
- Promote meetings prior to the use of the equipment.
- Ensure that the samples brought by the users are in perfect conditions of use, taking care of the
quality of the results generated. - Verify that the materials brought by the users are compatible with the equipment.
- Send the results obtained to users.
- Prepare guidelines on the use of equipment and materials.
- Supervise the packaging of the material for common use, as well as the maintenance of the
equipment. - Monitor the organization, operation, and management of stocks of commonly used materials.
- Provide for the repair of the equipment, complying with the rules in force in the ICBAS-UP.
- Ensure the other tasks necessary for the proper and efficient functioning of the UMIB Core Facility.
The provision of services by UMIB Core Facility may be contracted by a specific contract.
UMIB Core Facility will periodically promote surveys and other measures to assess the quality of services provided and levels of user satisfaction.