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| Isabel Nunes Correia |
| Technician responsible |
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Ph. |
+351 239 820190 |
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+351 239 822776 |
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| Location |
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Center for Neuroscience and Cell Biology of Coimbra
Faculty of Medicine
1st floor
Room 15
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| How to access our resources |
| FACSCalibur (Becton Dickinson) - 4 colours |
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| Software |
| CellQuest |
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| Flow cytometry principles |
Flow cytometry is a powerful technology for interrogating the phenotype and characteristics of cells. It simultaneously measures and analyzes multiple physical characteristics of single particles, usually cells, as they move, one by one in a fluid stream, through a beam of light and past sensors that measure physical and chemical characteristics of those particles or cells.
Analysis and differentiation of cells is based on size, granularity, and whether the cell is carrying fluorescent molecules in the form of either antibodies or dyes. As the cell passes through the laser beam, light is scattered in all directions. The light scattered in the forward direction at low angles (0.5-10º) from the axis is proportional to the square of the radius of a sphere and therefore to the size of the cell or particle. Light may enter the cell and be reflected and refracted by the nucleus and other contents of the cell, thus, the 90º light (right-angled, side) scatter may be considered proportional to the granularity of the cell.
The cells may be labelled with fluorochrome-linked antibodies or stained with fluorescent membrane, cytoplasmic, or nuclear dyes. Thus, differentiation of cell types, the presence of membrane receptors and antigens, membrane potential, pH, enzyme activity, protein expression, and DNA content may be measured.
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| How to use |
| First-time investigators must consult with the core technician concerning facility operating policies, and to discuss projects pursuant to optimizing experimental design, sample preparation, appropriate controls, and containment of potential pathogens. The facility cannot accept potentially pathogenic samples and if you have questions regarding viable human/primate samples or lentivirus-treated samples, please consult with the core technician. Samples labelled with radioactive materials are not accepted under any circumstances. |
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| How to access our resources |
First of all you should read the flow cytometry usage policy, this are some simple rules that you should follow when using the flow cytometer.
The second step will be to use our booking system in order to reserve some time on the system.
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| Flow Cytometry Usage Policy |
The first flow cytometry session for a new user must be attended by the responsible tecnhician.
Before using the flow cytometer, the user must have a valid reservation in the booking system. Check the Flow Cytometry Booking Policy for more information.
If you find a problem with the flow cytometer or evidence of incorrect usage immediately contact the responsible technician.
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| Flow Cytometry Booking Policy |
Booking changes must be made until one day before the booking time.
Users must always use only their own reserved time and should not use another user account to book time.
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| Reagents provided by the Flow Cytometry Unit |
| All users can freely use 12 x 75-mm polystyrene BD FalconTM tubes, BD FACSFlowTM sheath solution, BD FACSTM Clean or 10% household bleach cleaning solution and rinsing BD FACSTM solution. All fluorochromes, dyes and other reagents must be provided by the users. |
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| FACSCalibur Information |
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