Summary of the technology
Researchers of the University of Granada have developed probes based on nanoparticles, both fluorescent and functionalized with metals, which act as dual probes, and can be used for both flow cytometry (FACS –Fluorescent-Activated Cell Sorting) and mass cytometry (CyTOF -Cytometry by Time-Of-Flight-).
The probes can be used for bar coding of living cells or carrying out in vivo studies of different cell lineages through cell co-cultures as well as the activation of drug precursors through cytoplasmic catalysis of organometallic reactions inside living cells.
Description of the technology
The invention depicts the next aspects:
1. Dual Probes
The probes comprise nanoparticles both fluorescent and functionalized with metals which are suitable for carrying out both flow cytometry (FACS) and mass cytometry (CyTOF).
The nanoparticles are functionalized with a fluorophore so they are converted into fluorescent probes, as well as with metallic isotopes (e.g. palladium). The specific combination of a Fluorophore and specific metallic isotope confer a dual feature, allowing its detection through flow cytometry (fluorophore) and mass cytometry (metallic isotope).
These nanoparticles enter mammalian cells without a previous permeabilization step, without showing cytotoxic effects. These unique features open the option to do studies over living cells.
2. Cell codification procedure
The invention allows the cell codification through barcoding.
The living cells, through a nanofection process (introducing nanoparticles in cells), incorporate the metallic-fluorescent probes inside, which have a unique and predefined combination. Due to that specific combination, which is detected through mass cytometry and flow cytometry, the probes can act as bar codes.
In that way, a simultaneous reading of different cell lines can be made through cell co-cultures, since each cell line will have a specific and unique combination of isotopes and fluoride.
3. Synthesis procedure
- Emulsion and polymerization of a material that comprises a monomer conformed by heterocarbonated chains, a crossing agent, a polymerization initiator and a monomer that comprises at least an amine group or a carboxylic group, preferably amine, in order to obtain a crossed and functionalized nanoparticle.
- Conjugation among nanoparticles amine groups resulting from the previous step with the acid groups of the fluorides through solid-phase reactions.
- Incorporation of metals through metallic ions chelation with the fluorides.
Advantages and Benefits
- The nanoparticles can be used for both mass cytometry (CyTOF) and flow cytometry (FACS)
- It allows to carry out studies in living cells. The nanoparticles enter the cell without a previous permeabilization.
- The nanoparticles are not toxic, and do not affect the proteome.
- It is possible to carry out different cell lines co-cultures, multiple assays and subsequent barcoding.
- It allows the activation of drug precursor through cytoplasmic catalysis of organometallic reactions inside living cells.
- It is one of the first reagents for mass cytometry.
Current development status
Desired business relationship
Intellectual property status
Patent already applied for
University of Granada (OTRI)
Technology Transfer Office