Cellular biology unit: Microscopy and flow cytometry

Developed by: Instituto de Investigación Sanitaria Fundación Jiménez Díaz

Description

Cytometry and confocal microscopy have a broad array of applications spanning fields from biomedicine to the study of foods and materials, etc. Thanks to the development of new fluorescent probes and novel systems of detection, constant growth is being seen in the field studied and new applications are constantly appearing. Biomedical researchers at the IIS-FJD can perform 3-D studies on full or partial mountings, colocalization of fluorescent probes, tracing of molecules or ions within live or fixed cells, measurement of cells and organelles, and volume measurements. 

The tasks carried out in the unit are: 

  • Maintenance and calibration of the service's equipment;
  • Machine-use management for accounting and billing; 
  • Training courses and supervisory activities on proper use of analytical cytometry equipment; 
  • Maintenance and distribution of commonly used reagents for flow cytometry; 
  • Procurement of samples for FACScalibur and FACSCanto flow cytometry; 
  • Analysis of results obtained from different specialized software programs; 
  • Advisory on numerous complex techniques and protocols, such as which flourochromes are most appropriate for the different assays or which culture media are most appropriate for the different cell types; 
  • Advisory on making templates for flow cytometry; 
  • Reconstruction of images using multicolor software and 3-D imaging; and
  • Acting as a conduit between researchers and the applications services of equipment manufacturers. 

IIS-FJD has two flow cytometers and two Leica confocal microscopes. These are currently the institution's two most importantly and widely used research apparatuses. 

Common uses of flow cytometry: 

  • Immunophenotyping 
  • Cell-cycle analysis 
  • Reporter genes 
  • Cell proliferation assay 
  • Apoptosis 
  • Calcium flux assay 
  • Intracellular signaling: single-cell phospho-protein analysis 
  • Multiplex assay of molecule solution
  • Uses of confocal microscope:
  • Collection of sequences in series
  • 3-D reconstruction of objects, such as skin 
  • Measurement of marker colocalization 
  • Lateral images of the object 
  • Endocytosis