High throughput system for isolation, growth, and detection of lipid inclusions in bacteria
View Patent ↗A method of high throughput growth and quantitative analysis of microorganisms is provided that includes providing a microtiter plate growth and gas delivery system having well plates disposed for growth of the microorganisms, and providing a spectroscopic screening system disposed to analyze lipid inclusions of the microorganisms.
1. A method of providing a high throughput growth and quantitative analysis of microorganisms, comprising:
a. screening a throughput of cultures for bioplastic enrichment from an array of lipid producing bacteria using a microtiter plate growth and gas delivery system, wherein said microtiter plate growth and gas delivery system comprises well plates disposed for growth of microorganisms and a spectroscopic screening system disposed for analyzing said bioplastic and a gas delivery system comprising controlled gas flow of methane, nitrogen, oxygen, carbon dioxide, carbon monoxide, hydrogen, ethane, propane, or other gaseous hydrocarbons as a continuous gradient along the microtiter plate;
b. isolating lipid producing bacteria of said bioplastic;
c. inducing production of bioplastic;
d. determining a growth condition for enriching bioplastic production using said spectroscopic screening of said microtiter plate growth and gas delivery system.
2. The method according to claim 1 , wherein said spectroscopic screening system comprises a fluorescent screening system.
3. The method according to claim 2 , wherein said fluorescent screening comprises lipophilic fluorescent stains.
4. The method according to claim 3 , wherein said lipophilic fluorescent stain comprises Nile Red.
5. The method according to claim 2 , wherein said fluorescent screening comprises a flow cytometry screening system.
6. The method according to claim 2 , wherein said fluorescent screening comprises a plate reader screening system.
7. The method according to claim 1 , wherein said microorganisms are selected from the group consisting of bacteria, eukaryota, and archaea.
8. The method according to claim 7 , wherein said bacteria comprises methanotrophic.
9. The method according to claim 1 , wherein said microorganisms are grown in a pure culture.
10. The method according to claim 1 , wherein said microorganisms are grown in a mixed culture.
11. The method according to claim 10 , wherein mixed culture contains at least two known pure cultures.
12. The method according to claim 1 , wherein said microorganisms are grown in an enrichment culture.
13. The method according to claim 12 , wherein said enrichment culture contains an unknown consortium of microorganisms.
14. The method according to claim 13 , wherein the source of said microorganisms selected from the group consisting of soil, activated sludge, and other natural environments.
15. The method according to claim 1 , wherein said concentration gas gradient of gas constituents comprises a controlled gas mixture that is delivered to said plates.
16. The method according to claim 1 , wherein said lipid inclusions comprise biological polyesters.
17. The method according to claim 16 , wherein said biological polyesters comprise polyhydroxyalkanoates (PHAs).
18. The method according to claim 17 , wherein said PHAs comprise polyhydroxybutyrate (PHB).
19. The method according to claim 1 , wherein said microtiter plate growth and gas delivery system comprises a light source disposed to culture photosynthetic organisms.