Cytometry and Confocal Microscopy

The Institute of Plant Biochemistry and Photosynthesis (IBVF) provides support for the use of two flow cytometers and a confocal laser fluorescence microscope owned by the IBVF. This service includes initialising and running the equipment as well as technical assistance.

Those who use this confocal microscopy andflow cytometry service work mainly in the fields of biology and medicine, although occasionally some studies in materials science have used the confocal microscope.

Confocal microscopy can be used to analyse very diverse types of samples. In our facilities, it is generally used to analyse in vivo cultures of cyanobacteria or microalgae, vegetal tissues (leaves, roots and stems) or entire seedlings as well as cut sections of vegetal and animal tissues, and metal surfaces. 

The most common types of studies in this regard are as follows:

  • In vivo protein subcellular localisation detected via fusion with fluorescent proteins (e.g., GFP, YFP, CFP)
  • Cell expression studies employing promoter fusion with reporter genes of fluorescent proteins
  • BIFC studies
  • Detection of inmunocytochemical markers in tissues
  • Colocalisation analyses
  • Obtention of serial sections for 2D and 3D pseudo-reconstruction

The samples that are generally analysed using our flow cytometry service are cell suspensions (microalgae, cyanobacteria, vegetal or animal cells) in aqueous solution. 

The most common applications in this regard are as follows:

  • Size distribution and cellular complexity analyses
  • Detection of specific cellular types by marking them with different fluorochromes (the mark can be made using antibodies that recognise surface antigens or by expressing fluorescent proteins inside of the cells)
  • Cell cycle analyses (of specific populations) via the quantification of the amount of DNA present in each cell
  • Separation of up to four cell populations at once, which is different from a cell suspension (the separation can be performed in different types of laboratory tubes as well as in different types of multiwell plates)
  • Production of cell clones by the separation of individual cells