1. Centrifuges By Magnus Manske (Own work) [CC BY 1.0 (http://creativecommons.org/licenses/by/1.0), GFDL (http://www.gnu.org/copyleft/fdl.html) or CC-BY-SA-3.0 (http://creativecommons.org/licenses/by-sa/3.0/)], via Wikimedia Commons

2. Quiz Clinical Use History Specifications Principles of Operation Block Diagram Commercial Examples Preventive Maintenance Common Problems Test Procedures Summary

3. 1707-1751 - English engineer Benjamin Robins invented a whirling arm apparatus to determine drag 1864 - Antonin Prandl invented the first dairy centrifuge in order to separate cream from milk History Wikitanvir (2010), Portrait of Benjamin Robins [painting]. Reproduction courtesy of the Supreme Court Historical Society

4. 1879 - Gustaf de Laval demonstrated the first continuous centrifugal separator 1926 Theodor Svedberg Nobel Prize for invention of the ultracentrifuge 1940 - SPINCO produced the first commercial ultracentrifuge History Celsius~commonswiki (2005), Gustaf de Laval [photograph]. Retrieved from https://commons.wikimedia.org/wiki/File:GustafD eLaval.jpg Unknown Author (1929), Theodor Svedberg [photograph]. Retrieved from https://commons.wikimedia.org/wiki/File:The- svedberg-1.jpg

5. Clinical Use Separating plasma from the blood suspension Wikipedia. “Laboratory Centrifuge.” Wikipedia, p. 1-12. Retrieved from: https://en.wikipedia.org/wiki/ Laboratory_centrifuge

6. Separate substances of different densities Make a counterbalance for the centrifuge tube you want to put in the centrifuge Principles of Operation Occupational Safety and Health Administration. “Laboratory Safety: Centrifuges.” OSHA. Last update: August, 2011. Retrieved from: https://www.osha.gov/Publications/laboratory/OSHAquickfacts-lab- safety-centrifuges.pdf

7. Principles of Operation The acceleration is measured in multiples of "g“, the standard acceleration due to gravity at the Earth's surface, or × "g", and it is given by where g = acceleration r = rotational radius (centimeter, cm) N = rotating speed (revolutions per minute, r/min) RCF=rational canonical form

8. ManualElectrical Principles of Operation Wikipedia. “Laboratory Centrifuge.” Wikipedia, p. 1-12. Retrieved from: https://en.wikipedia.org/wiki/ Laboratory_centrifuge Nadina Wiórkiewicz (2011), Beckman SW28 centrifuge rotor [photograph]. Retrieved from https://commons.wikimedia.org/wiki/File :Beckman_SW28_centrifuge_rotor_1.jpg

9. Principles of Operation Types: - Table Top - Floor Mount - Micro centrifuge (hematocrit) - Refrigerated - Cell Washer

10. Separation Methods - Centrifuges Types of Separation Differential Isopycnic Rate Zonal Types of Centrifuges Ultracentrifuge (1000000g = 9800 km/s²) - Analytical - Preparative High Speed Table Top Clinical Microfuges Wikipedia. “Laboratory Centrifuge.” Wikipedia, p. 1-12. Retrieved from: https://en.wikipedia.org/wiki/ Laboratory_centrifuge

11. Separation Methods - Centrifuges Differential (separates liquids from solids) Thomasione (2009), Differential Centrifugation [drawing]. Retrieved from https://commons.wikimedia.org/wiki/File:Differentielle_zentrifugation.png

12. Separation Methods - Centrifuges Isopycnic (separation based upon particle density) Virginia Reid (2015) Isopycnic Separation [diagram].

13. Separation Methods - Centrifuges Rate Zonal (separation based upon particle size) Virginia Reid (2015) Rate Zonal [diagram].

14. Centrifuge RCF Classification

15. Common User Problems 1.Incorrect Loading or Balancing 2.Incorrect Attachment 3.Consumable Failure 4.Overloading 5.Corrosion WHO. “Chapter 7: Centrifuge.” From the publication: Maintenance Manual for Laboratory Equipment, WHO: 2008, pgs. 45-49.

16. Basic parts: –Speed motor –Timer (mechanical or electronic) –Rotor –Lid interlock (after 1990) Principles of Operation

17. WHO. “Chapter 7: Centrifuge.” From the publication: Maintenance Manual for Laboratory Equipment, WHO: 2008, pgs. 45-49.

18. Principles of Operation WHO. “Chapter 7: Centrifuge.” From the publication: Maintenance Manual for Laboratory Equipment, WHO: 2008, pgs. 45-49.

19. Low speed centrifuges - up to 12,000 RPM High speed - 35,000 RPM (most refrigerated) Ultrahigh speed - 125,000 RPM (most refrigerated) Principles of Operation

20. 4. Centrifuge filtrate at low speeds ( 1000 X g for 10mins ) This pellets the nuclei as this is the densest organelle Principles of Operation BruceBlaus (2013), Nucleus [drawing]. Retrieved from https://commons.wikimedia.org/wiki/File:Blausen_0212_CellNucleus.png

21. 5. Centrifuge at medium speeds ( 10 000 x g for 30 mins ) This pellets mitchondria which are the second densest organelle Principles of Operation Nevit (2010), Mitochondria [drawing]. Retrieved from https://commons.wikimedia.org/wiki/File:Mitochondria.svg

22. 6. Centrifuge at high speeds ( 100 000 x g for 30 mins) This pellets ER, golgi apparatus and other membrane fragments Principles of Operation BruceBlaus (2013), Golgi Apparatus [drawing]. Retrieved from https://en.wikipedia.org/wiki/Golgi_apparatus

23. 7 Centrifuge at very high speeds ( 300 000 x g for 3hrs) This pellets ribosomes Principles of Operation Vossman (2009), Ribosomes [image]. Retrieved from https://commons.wikimedia.org/wiki/File:Ribosome_shape.png

24. Controls Speed Timer Temperature

25. Commercial Examples Wikipedia. “Laboratory Centrifuge.” Wikipedia, p. 1-12. Retrieved from: https://en.wikipedia.org/wiki/ Laboratory_centrifuge

26. Commercial Examples Matylda Sęk (2011), Beckman-Coulter preparative centrifuge [photograph]. Retrieved fromhttps://commons.wikimedia.org/wiki/File:Beckman- Coulter_preparative_centrifuge_Avanti_J25-02.jpg

27. Commercial Examples 2015, Centrifuge [screenshot] retrieved from https://www.google.com/webhp?sourceid=chrome-instant&ion=1&espv=2&ie=UTF- 8#q=centrifuge&tbm=shop

28. Power supply (machine will not start) Lid closure switch- machine will not start Timer - bypass Rotor Motor (weakened, off balance/bent) -Not working properly -Making noise Lid does not open or close Common Problems

29. Troubleshooting Cooper, Justin and Alex Dahinten for EWH. “Centrifuge Troubleshooting Flowchart.” From the publication: Medical Equipment Troubleshooting Flowchart Handbook. Durham, NC: Engineering World Health, 201

30. Run a cycle with empty/balanced buckets or test tube holders. Test the motor speed (fluorescent bulb 60 Hz) Test Procedures

31. Safety Do not bypass interlock! Centrifuge should be de-contaminated before working on it Never attempt to touch or stop a spinning rotor by hand or with another tool or object If handling blood or body fluids, use protection (gloves, mask, gown, etc.)

32. Lubricate and clean motor Clean case Ensure tightness of rotor Verify that alarms are operating properly Ensure interlock is functioning If refrigerated, ensure temperature reading is accurate. Replace/repair gaskets, seals, and vacuum pump (if applicable) Check commutator, bearings and brushes if applicable Always check the rotor for any cracks. A cracked rotor MUST be discarded. NEVER attempt repair. Preventive Maintenance

33. Questions ?