Cellular Biology Unit: Microscopy and flow cytometry
Role and objectives
In recent years, cell biology and microscopy and flow-cytometry techniques have advanced considerably within the research activity performed in the institute. Today, most all groups have incorporated this technology into their research.
For this reason, having a cellular biology unit will boost all of the research done in the IIS-FJD.
The unit's tasks shall be:
- Drafting SOPs
- Advising on the techniques and protocols in place for the correct use of the equipment
- Updating available protocols and new technology
- Aiding in the preparation and processing of images within the area of confocal microscopy
Once the unit has been set up and is operational, the goal is to turn it into another provider of training opportunities, organizing courses on the handling and operation of the equipment available in the unit. In time, habitual and potential users will be able to receive continuous training.
The overall objectives of the unit are therefore to standardize the techniques, handling, and use of equipment, thus leading to greater efficiency. Also, the unit intends to become a point of support available to researchers working in cellular biology. In this way, the IIS-FJD will increase and improve its scientific and technological facilities and will bolster the research capacity of groups working in R&D, increasing and improving the transfer of research results.
The Cellular Biology Unit will report to the Research Committee and will be led by one head of service. A proposal has been made to hire a qualified, university-educated technician for this spot. The candidate is currently Beatriz Sastre Turrión.
At the moment, a part-time technician is working within the area of confocal microscopy, although a mid-level support technician has been requested.
The Fundación Jiménez Díaz currently features the following:
FACSCalibur flow cytometry system (Becton Dickinson)
Contains 2 air-cooled lasers (1 argon-ion laser (488 nm) and 1 solid-state red diode laser red diode laser (635 nm))
The device can analyze up to 4 colors and 8 parameters at a time (FSC, SSC, 4 colors, pulse processing, and time)
The FACSCalibur flow cytometer makes it possible to analyze samples labeled with the following fluorochromes:
- FL1: FITC, Alexa 488
- FL2: PE, PI
- FL3: PE-Cy5, PerCP, Per-CP 5.5
- FL4: APC, APC Cy7, Alexa 647
The cytometer has a Macintosh G4 JGH2 processor, a 40-GB hard drive, using the Cell Quest program for acquisition and analysis.
FACS Vantage SE cytometer-sorter, a state-of-the art cell analyzer and sorter.
The system contains a module for results analysis. This system includes:
- a. Power Mac G3
- b. Monitor
- c. HP DeskJet 1200 C/PS laser printer
- d. Software: Cell Quest version 3.4, FACS Assistant
2 Leica confocal microscopes (SP2 and SP5)
The latter has a microscope camera for in vivo studies
10 light microscopes and 4 fluorescence microscopes
Currently, the cytometry and confocal-microscopy unit is in two different areas of the same building.
The cytometry unit is on the 4th floor of the IIS-FJD. Both the sorter and the cytometer are in two different rooms in order to insure the most appropriate conditions for both the equipment and the type of experiments that they are used to carry out.
The confocalmicroscopy unit is on the 2nd floor of the same building. Both microscopes are in a room that is specifically equipped for them, especially in terms of temperature, as this is one of the most crucial factors for microscopes of these characteristics to function properly.
Access and safety regulations
Researchers can use both services by contacting the unit head to request a session.
These sessions will be assigned based on equipment availability during the operating hours of the unit. Groups requiring longer sessions than normal may request a new slot.
Users of the service must keep to their allotted slots, taking into account the time needed to clean the equipment (in the case of flow cytometry); also, users must take care to follow the instructions for turning the machines on and off.
The hospital's internal regulations on biosafety must be followed.