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Growing and Harvesting Of Bacterial Culture
- 1. • Growing And Harvesting Of Bacterial Culture By Muhammad Ramzan To Dr.kashif Rahim
- 2. Culture Media Used to grow bacteria Can be used to: Enrich the number of bacteria Increase The production of certain bacteria and suppress others. It defferentiate among different kind of bacteria
- 3. Purpose of Culturing To create antigen for laboratory use Estimating viable count Maintan stock culture Isolation Properties of bacteria
- 4. Method of Isolation Of Pure Culture Surface Plating Enrichment medium Selective medium
- 5. Type of Media Used Selective Media:- Favour the growth of Particular microorganisms And inhibit the growth of others Differential Media:-Distinguish between different of bacteria on the basis of their characters
- 6. Types Of Media Used Enrichment Media:- Suplemented by blood Or other special nutrients or encourage the growth Fastidious Heterotroph General purpose Media:- Will support the Growth of many microorganisms
- 7. Culturing Used to grow bacteria Enrich the number of Bacteria Differentiate among different kind of bacteria
- 8. Method To Isolate The Bacteria Streak Culture Liquid Culturing Stab Pour plate
- 9. How To Inoculate Bacteria Sample On Plate Plate:- Provide large Surface For Isolation and observation Of colonies Using a sterile loop Or a sterile swab.streak Your sample on petri plate But first Cool sterilized loop Before picking up sample
- 10. Macconkey Agar Macconkey agar Has colour indicator that distinguish presence Of Acid Bacteria that ferments a particular sugar will produce acid Wastes on plates,turn pH indicator red
- 11. Streak Culture Lawn or carpet culture to create uniform suface of microorganisms Bacteriophage typing To obtain large number of antigens
- 12. Liquid Culturing Liquid culture are done in Tubes Bottles Flask Blood culture Water analysis
- 13. Stab Culture Puncturing Suitable medium Such as nutrients,agar,gelatin Observe Gelatin liquefaction Preserving the Stock culture
- 14. Incubation After streaking,place the sample in incubator at 37°C for 24-48 hours After 24-48 hours we observe the Colonies of bacterial cell
- 15. Harvesting Harvesting done for Refreshing culture medium or split Cultures into more Cultures Dushes or flasks It takes several steps 1.Cell detachment 2 .Depth Filtration 3.Centrifugation
- 16. Cell Detachment Before centrifugation Or Filtration cells that adhere First may be detached Manual cell Detachment By scraping Is quick but may compromise Cell viability And may not be feasible On an industrial scale Trypsin:-A protease enzyme comm only used in lab For this purpose work by cleaving the amino acid
- 17. Cell Detachment Non-enzymatic Treatment like EDTA Or citric saline are gentle Detachment treatment
- 18. Centrifugation For bacterial Cells that are Grown in suspenSion Centrifugation can be the first step in cell harvesting There need to be enough centrifugal force To cause the dense solid Cell to collect At the bottom of centrifugation tube But not much to destroy cell membrane
- 19. Disk stack centrifuge Its originally develop for the dairy industry to separate Fat from milk without shearing the fat particles,have been applied to bacterial cells havervesting Because its gentle On bacterial cell,which are particularly.
- 20. Depth Filtration Depth Filters are made of a porous Filtration medium that traps particles or cells throughout the medium rather then Particles remaining on surface only Depth filtration can Replace centrifugation As a method of capturing Bacterial cells Or can be used as a second step Filters are flushid with buffers To remove loose particles andfulished to recover the Bound cells