EIJ_SWS15_FLRed6_auto_uls_phyto_600s 2018-11-23 14h02.cyz ----- Processed at : 23-Nov-2018 14:13:04 ----- 1. The cytometric analysis is performed on machine Eijsden 2. The CytoUSB software used is CytoUSB Version: 5.7.9.20 3. The filename is EIJ_SWS15_FLRed6_auto_uls_phyto_600s 2018-11-23 14h02.cyz 4. The analysis is performed on 23-11-2018 time 14:02:49 5. The number of measured particles is 566 6. The total concentration found (e.g. only SWS trigg.) is 70 particles/microliter 7. The concentration found (all used triggers) is 1 particles/microliter 8. The preconcentration (before analysis) found (all used triggers) is 0 particles/microliter 9. The flush time found (between samples) is 130 seconds 10. The effective analysis volume is 1037.8424 microliter 11. The pumped volume is 2748 microliter 12. The particle-rate is 320 particles passing the laser beam/second 13. The used sample flow rate is 4.5842 microliter/second 14. The used block size memory is 256 kB 15. Trigger 1: SWS HS at threshold level 15 mV 16. Trigger 2: Maximum FL Red HS at threshold level 6 mV 17. Temperature sensor PMT is : 30 C 18. Temperature sensor Sheath is : 26 C 19. Temperature sensor Sheath is : 30 C 20. Pressure sensor difference is : -207 mBar 21. Pressure sensor absolute is : 247 mBar 22. Sample pump 1: Last calibration of the sample pump was : 02-Aug-2018 23. Alignment check 1 by curvature detector is 1.0608 (1=perfect) 24. Alignment check 2 by curvature detector is 59% (100%=perfect) 25. Saturated signals (flattened peaks) occur when the dynamic range of the (HS)detector is reached 26. FWS: Nr of saturated signals is 1 (0% of 566 measured particles) 27. FWS: Maximum signal height found is 9911 mV 28. FWS: Nr of particles with signal height <10 mV is high (29 part. (5 %)) 29. SWS HS: Nr of saturated signals is 0 (0% of 566 measured particles) 30. SWS HS: Maximum signal height found is 3745 mV 31. SWS HS: Nr of particles with signal height <10 mV is high (2 part. (0 %)) 32. FL Yellow HS: Nr of saturated signals is 0 (0% of 566 measured particles) 33. FL Yellow HS: Maximum signal height found is 57 mV 34. FL Yellow HS: Nr of particles with signal height <10 mV is high (491 part. (87 %)) 35. FL Orange HS: Nr of saturated signals is 0 (0% of 566 measured particles) 36. FL Orange HS: Maximum signal height found is 72 mV 37. FL Orange HS: Nr of particles with signal height <10 mV is high (516 part. (91 %)) 38. FL Red HS: Nr of saturated signals is 0 (0% of 566 measured particles) 39. FL Red HS: Maximum signal height found is 124 mV 40. FL Red HS: Nr of particles with signal height <10 mV is high (448 part. (79 %)) 41. FWS: Nr of block shaped signals is 151 (26.6784%) 42. SWS HS: Nr of block shaped signals is 52 (9.1873%) 43. FL Yellow HS: Nr of block shaped signals is 23 (4.0636%) 44. FL Orange HS: Nr of block shaped signals is 9 (1.5901%) 45. SWS LS: Nr of block shaped signals is 1 (0.17668%) 46. Suspect nr: 0=low, 10=high 47. Maximum FWS: suspect = 4 48. Maximum SWS HS: suspect = 4 49. Maximum FL Yellow HS: suspect = 2 50. Maximum FL Orange HS: suspect = 3 51. Maximum FL Red HS: suspect = 4 52. Maximum SWS LS: suspect = 0 53. Maximum FL Yellow LS: suspect = 2 54. Maximum FL Red LS: suspect = 0 55. SWS HS: PMT level is 56 56. FL Yellow HS: PMT level is 79 57. FL Orange HS: PMT level is 78 58. FL Red HS: PMT level is 97 59. SWS LS: PMT level is 40 60. FL Yellow LS: PMT level is 64 61. FL Red LS: PMT level is 73 -------------------------- Warnings info -------------------------- 1.01 Warning: The number of saved particles in this analysis is quite low and 1.01 Warning: you might miss (statistically) bigger particles in your sample 1.01.1 Solution - Increase number of measured particles. Increase setted time OR after setted number of analyzed particles (in CytoUSB) 1.03 Warning: Your preconcentration differs more than 25% from the during analysis concentration 1.03.1 Solution - Carry over between samples? > Flush longer 1.03.2 Solution - Better homogenize samples during subsampling by a gentle stirrer 1.03.3 Solution - Colonies-filaments of cells break during analysis > stir slower 1.05 Warning: Your effective analysis volume is low compared to the pumped analysis volume 1.05.1 Solution - Set blocksize memory in CytoUSB higher, used=256k (default=4k) e.g. to 512k 1.17 Warning: Last calibration of sample pump towards analysis date was more than 90 days. (02-Aug-2018) 1.17.1 Solution - Check if the pumped analysis volume is the real volume by weighing 1.19 Warning: FWS-L and -R might indicate an air bubble problem 59% (100%=perfect) 1.19.1 Solution - Pinch valve to get rid of the air bubble or blockage by something different 1.20 Warning: SWS LS: HS-LS ratio is 29 and too low. 1.20.1 Solution - Put LS PMT lower. Ratio HS/LS should be around 100. 1.20 Warning: FL Yellow LS: HS-LS ratio is 999 and too high. 1.20.1 Solution - Put LS PMT higher. Ratio HS/LS should be around 100. 1.20 Warning: FL Yellow LS: HS-LS ratio is 999 and too high. 1.20.1 Solution - Put LS PMT higher. Ratio HS/LS should be around 100. 1.20 Warning: FL Red LS: HS-LS ratio is 999 and too high. 1.20.1 Solution - Put LS PMT higher. Ratio HS/LS should be around 100. 1.22 Warning: FL Yellow HS: Biggest particles found is only 57 mV. Detector sensitivity might be too low. 1.22.1 Solution - Increase, set higher FL Yellow HS PMT voltage in CytoUSB (make more sensitive) 1.23 Warning: FL Yellow HS : Nr of particles with signal height <10 mV is high (87 %) 1.23.1 Solution - Increase, set higher trigger threshold (to prevent measuring noise) 1.23.2 Solution - Decrease, set lower particle sample rate. Maybe caused by an overload of particles/ noise in your sample 1.23.3 Solution - Clean sheath, clean cuvette, clean tubing? (noise can be introduced by dirty walls) 1.22 Warning: FL Orange HS: Biggest particles found is only 72 mV. Detector sensitivity might be too low. 1.22.1 Solution - Increase, set higher FL Orange HS PMT voltage in CytoUSB (make more sensitive) 1.23 Warning: FL Orange HS : Nr of particles with signal height <10 mV is high (91 %) 1.23.1 Solution - Increase, set higher trigger threshold (to prevent measuring noise) 1.23.2 Solution - Decrease, set lower particle sample rate. Maybe caused by an overload of particles/ noise in your sample 1.23.3 Solution - Clean sheath, clean cuvette, clean tubing? (noise can be introduced by dirty walls) 1.22 Warning: FL Red HS: Biggest particles found is only 124 mV. Detector sensitivity might be too low. 1.22.1 Solution - Increase, set higher FL Red HS PMT voltage in CytoUSB (make more sensitive) 1.23 Warning: FL Red HS : Nr of particles with signal height <10 mV is high (79 %) 1.23.1 Solution - Increase, set higher trigger threshold (to prevent measuring noise) 1.23.2 Solution - Decrease, set lower particle sample rate. Maybe caused by an overload of particles/ noise in your sample 1.23.3 Solution - Clean sheath, clean cuvette, clean tubing? (noise can be introduced by dirty walls) 1.24 Warning: FWS: A relative large amount of particles are (rather) block shaped (27%) 1.24.1 Solution - Decrease, set lower particle sample rate. Maybe caused by an overload of particles/ noise in your sample 1.24.2 Solution - Decrease, set lower FWS PMT voltage in CytoUSB (make less sensitive) 1.24 Warning: SWS HS: A relative large amount of particles are (rather) block shaped (9%) 1.24.1 Solution - Decrease, set lower particle sample rate. Maybe caused by an overload of particles/ noise in your sample 1.24.2 Solution - Decrease, set lower SWS HS PMT voltage in CytoUSB (make less sensitive) 1.25 Warning: Suspected particles data blocks found! 1.25.1: Maximum FWS: suspect = 4 1.25.1 Solution - Particle rate too high? 1.25.1 Solution - Trigger too sensitive? 1.25.1 Solution - Details look at 'DATA DIAGNOSIS' in 'Check CytoSense-Diagnosis-Meta Data' 1.25.2: Maximum SWS HS: suspect = 4 1.25.2 Solution - Particle rate too high? 1.25.2 Solution - Trigger too sensitive? 1.25.2 Solution - Details look at 'DATA DIAGNOSIS' in 'Check CytoSense-Diagnosis-Meta Data' 1.25.3: Maximum FL Orange HS: suspect = 3 1.25.3 Solution - Particle rate too high? 1.25.3 Solution - Trigger too sensitive? 1.25.3 Solution - Details look at 'DATA DIAGNOSIS' in 'Check CytoSense-Diagnosis-Meta Data' 1.25.4: Maximum FL Red HS: suspect = 4 1.25.4 Solution - Particle rate too high? 1.25.4 Solution - Trigger too sensitive? 1.25.4 Solution - Details look at 'DATA DIAGNOSIS' in 'Check CytoSense-Diagnosis-Meta Data'