EIJ2_SWS15_FLRed6_auto_uls_IMG_600s 2021-11-30 12h04.cyz ----- Processed at : 30-Nov-2021 12:15:29 ----- 1. The cytometric analysis is performed on machine LelystadII 2. The CytoUSB software used is CytoUSB Version: 5.9.1.5 3. The filename is EIJ2_SWS15_FLRed6_auto_uls_IMG_600s 2021-11-30 12h04.cyz 4. The analysis is performed on 30-11-2021 time 12:05:01 5. The number of measured particles is 98568 6. The total concentration found (e.g. only SWS trigg.) is 486 particles/microliter 7. The concentration found (all used triggers) is 181 particles/microliter 8. The preconcentration (before analysis) found (all used triggers) is 181 particles/microliter 9. The flush time found (between samples) is 110 seconds 10. The effective analysis volume is 545.1261 microliter 11. The pumped volume is 1218 microliter 12. The particle-rate is 988 particles passing the laser beam/second 13. The used sample flow rate is 2.0349 microliter/second 14. The used block size memory is 4 kB 15. Trigger 1: SWS HS at threshold level 15 mV 16. Trigger 2: Maximum FL Red HS at threshold level 6 17. Temperature sensor PMT is : 26 C 18. Temperature sensor Sheath is : 22 C 19. Temperature sensor Sheath is : 27 C 20. Pressure sensor difference is : -788 mBar 21. Pressure sensor absolute is : 667 mBar 22. Sample pump 1: Last calibration of the sample pump was : 08-Sep-2021 23. Alignment check 1 by curvature detector is 1.1061 (1=perfect) 24. Alignment check 2 by curvature detector is 47% (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 98568 measured particles) 27. FWS: Maximum signal height found is 6518 mV 28. FWS: Nr of particles with signal height <10 mV is high (63808 part. (65 %)) 29. SWS HS: Nr of saturated signals is 0 (0% of 98568 measured particles) 30. SWS HS: Maximum signal height found is 2659 mV 31. SWS HS: Nr of particles with signal height <10 mV is high (76 part. (0 %)) 32. FL Yellow HS: Nr of saturated signals is 0 (0% of 98568 measured particles) 33. FL Yellow HS: Maximum signal height found is 3216 mV 34. FL Yellow HS: Nr of particles with signal height <10 mV is high (7684 part. (8 %)) 35. FL Orange HS: Nr of saturated signals is 0 (0% of 98568 measured particles) 36. FL Orange HS: Maximum signal height found is 3487 mV 37. FL Orange HS: Nr of particles with signal height <10 mV is high (37752 part. (38 %)) 38. FL Red HS: Nr of saturated signals is 3 (0% of 98568 measured particles) 39. FL Red HS: Maximum signal height found is 5000 mV 40. FL Red HS: Nr of particles with signal height <10 mV is high (96705 part. (98 %)) 41. FL Yellow HS: Nr of block shaped signals is 10047 (10.193%) 42. FL Orange HS: Nr of block shaped signals is 606 (0.6148%) 43. SWS HS: Nr of block shaped signals is 134 (0.13595%) 44. FWS: Nr of block shaped signals is 8 (0.0081162%) 45. FL Red HS: Nr of block shaped signals is 7 (0.0071017%) 46. SWS LS: Nr of block shaped signals is 1 (0.0010145%) 47. Suspect nr: 0=low, 10=high 48. Maximum FWS: suspect = 0 49. Maximum SWS HS: suspect = 0 50. Maximum FL Yellow HS: suspect = 0 51. Maximum FL Orange HS: suspect = 3 52. Maximum FL Red HS: suspect = 2 53. Maximum SWS LS: suspect = 0 54. Maximum FL Yellow LS: suspect = 0 55. Maximum FL Red LS: suspect = 0 56. SWS HS: PMT level is 47 57. FL Yellow HS: PMT level is 88 58. FL Orange HS: PMT level is 86 59. FL Red HS: PMT level is 101 60. SWS LS: PMT level is 34 61. FL Yellow LS: PMT level is 65 62. FL Red LS: PMT level is 75 63. IIF: Imaging in Flow pictures available. Nr of images is 138 -------------------------- Warnings info -------------------------- 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=4k (default=4k) e.g. to 8k 1.09 Warning: This (default) low block size of 4kB in CytoUSB will decrease the effective analysis volume 1.09.1 Solution - Increase block size memory if the total particle rate is high enough (but prevent time outs) 1.15 Warning: Pressure difference is high. 1.15.1 Solution - Check if there is something blocking from sample inlet to sheath (filter block, tubing) 1.19 Warning: FWS-L and -R might indicate an air bubble problem 47% (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 34 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 33 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 32 and too low. 1.20.1 Solution - Put LS PMT lower. Ratio HS/LS should be around 100. 1.20 Warning: FL Red LS: HS-LS ratio is 64 and too low. 1.20.1 Solution - Put LS PMT lower. Ratio HS/LS should be around 100. 1.23 Warning: FWS : Nr of particles with signal height <10 mV is high (65 %) 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) 1.23 Warning: FL Red HS : Nr of particles with signal height <10 mV is high (98 %) 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: FL Yellow HS: A relative large amount of particles are (rather) block shaped (10%) 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 FL Yellow HS PMT voltage in CytoUSB (make less sensitive) 1.25 Warning: Suspected particles data blocks found! 1.25.1: Maximum FL Orange HS: suspect = 3 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'