EasyClus check & Info of last file ----- Processed at : 24-Apr-2025 06:27:02 ----- 1. The cytometric analysis is performed on machine Lobith 2. The CytoBuoy instrument serial-nr is CS-2021-106 3. The CytoUSB software used is CytoUSB Version: 5.11.3.1 4. LASER : OBIS LS 488-60 : Used at 50 mW 5. LASER : OBIS LS 552-60 : Used at 50 mW 6. The filename is EIJ2_SWS100_FLO100_2_uls_beads_600s 2025-04-24 06h12.cyz 7. The analysis is performed on 24-04-2025 time 06:16:07 8. The number of measured particles is 492 9. The total concentration found (e.g. only SWS trigg.) is 117 particles/microliter 10. The concentration found (all used triggers) is 1 particles/microliter 11. The preconcentration (before analysis) found (all used triggers) is 1 particles/microliter 12. The flush time found (between samples) is 130 seconds 13. The effective analysis volume is 849.8538 microliter 14. The pumped volume is 852 microliter 15. The particle-rate is 167 particles passing the laser beam/second 16. The used sample flow rate is 1.4245 microliter/second 17. Trigger 1: SWS HS at threshold level 100 mV 18. Trigger 2: Maximum Fl Orange at threshold level 100 19. Temperature sensor PMT is : 25.2 C 20. Temperature sensor Sheath is : 20.3 C 21. Temperature sensor Laser is : 24.6 C 22. Temperature sensor System is : 22.7 C 23. Pressure sensor difference is : -448 mBar 24. Pressure sensor absolute is : 342 mBar 25. Sample pump 1: Last calibration of the sample pump was : 05-Dec-2024 26. Alignment check 1 by curvature detector is 1.088 (1=perfect) 27. Alignment check 2 by curvature detector is 70.1% (>70%=perfect) 28. Saturated signals (flattened peaks) occur when the dynamic range of the (HS)detector is reached 29. FWS: Nr of saturated signals is 7 (0% of 492 measured particles) 30. FWS: Maximum signal height found is 14464 mV 31. FWS: Nr of particles with signal height <10 mV is high (0 part. (0 %)) 32. SWS HS: Nr of saturated signals is 1 (0% of 492 measured particles) 33. SWS HS: Maximum signal height found is 7232 mV 34. SWS HS: Nr of particles with signal height <10 mV is high (0 part. (0 %)) 35. FL Yellow HS: Nr of saturated signals is 1 (0% of 492 measured particles) 36. FL Yellow HS: Maximum signal height found is 7360 mV 37. FL Yellow HS: Nr of particles with signal height <10 mV is high (0 part. (0 %)) 38. FL Orange HS: Nr of saturated signals is 3 (0% of 492 measured particles) 39. FL Orange HS: Maximum signal height found is 7360 mV 40. FL Orange HS: Nr of particles with signal height <10 mV is high (0 part. (0 %)) 41. FL Red HS: Nr of saturated signals is 6 (0% of 492 measured particles) 42. FL Red HS: Maximum signal height found is 7360 mV 43. FL Red HS: Nr of particles with signal height <10 mV is high (0 part. (0 %)) 44. FWS: Nr of block shaped signals is 27 (5.4878%) 45. FL Yellow HS: Nr of block shaped signals is 9 (1.8293%) 46. FL Orange HS: Nr of block shaped signals is 9 (1.8293%) 47. FL Red HS: Nr of block shaped signals is 9 (1.8293%) 48. FL Red LS: Nr of block shaped signals is 7 (1.4228%) 49. SWS HS: Nr of block shaped signals is 5 (1.0163%) 50. Suspect nr: 0=low, 10=high 51. Maximum FWS: suspect = 4 52. Maximum SWS HS: suspect = 4 53. Maximum SWS LS: suspect = 0 54. Maximum FL Yellow HS: suspect = 3 55. Maximum FL Yellow LS: suspect = 0 56. Maximum FL Orange HS: suspect = 5 57. Maximum FL Red HS: suspect = 3 58. Maximum FL Red LS: suspect = 0 59. SWS HS: PMT level is 49 60. SWS LS: PMT level is 48 61. FL Yellow HS: PMT level is 94 62. FL Yellow LS: PMT level is 76 63. Fl Orange: PMT level is 105 64. FL Red HS: PMT level is 110 65. FL Red LS: PMT level is 81 66. IIF: Imaging in Flow pictures available. Nr of images is 250 -------------------------- 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) 1.11 Warning: This threshold level is quite high 1.11 Warning: Take care you don't miss smaller particles, because they will not be triggered at this level 1.11.1 Solution - Set lower threshold trigger level 1.11.2 Solution - If you need to put the level that high to measure something, it might be causeed by increased noise. 1.11.3 Solution - Increased noise is usually caused by dirty cuvette (inner) walls/tubing/sheath fluid - So, clean the cuvette carefully, change tubing, use new sheath 1.21 Warning: Last calibration of sample pump towards analysis date was more than 90 days. (05-Dec-2024) 1.21.1 Solution - Check if the pumped analysis volume is the real volume by weighing 1.24 Warning: SWS LS: Maximum is 340 mV if HS is around 4500 mV and too high. 1.24.1 Solution - Put LS PMT lower. Maximum LS around 50 mV (if HS is around 4500 mV). 1.24 Warning: SWS LS: HS-LS ratio is 67 and too low. 1.24.1 Solution - Put LS PMT lower. Ratio HS/LS should be around 100. 1.24 Warning: FL Yellow LS: HS-LS ratio is 55 and too low. 1.24.1 Solution - Put LS PMT lower. Ratio HS/LS should be around 100. 1.24 Warning: FL Yellow LS: HS-LS ratio is 46 and too low. 1.24.1 Solution - Put LS PMT lower. Ratio HS/LS should be around 100. 1.24 Warning: FL Red LS: HS-LS ratio is 68 and too low. 1.24.1 Solution - Put LS PMT lower. Ratio HS/LS should be around 100. 1.28 Warning: FWS: A relative large amount of particles are (rather) block shaped (5%) 1.28.1 Solution - Decrease, set lower particle sample rate. Maybe caused by an overload of particles/ noise in your sample 1.28.2 Solution - Decrease, set lower FWS PMT voltage in CytoUSB (make less sensitive) 1.25 Warning: Suspected particles data blocks found! 1.29.1: Maximum FWS: suspect = 4 1.29.11 Solution - Particle rate too high? 1.29.21 Solution - Trigger too sensitive? 1.29.31 Solution - Details look at 'DATA DIAGNOSIS' in 'Check CytoSense-Diagnosis-Meta Data' 1.29.2: Maximum SWS HS: suspect = 4 1.29.12 Solution - Particle rate too high? 1.29.22 Solution - Trigger too sensitive? 1.29.32 Solution - Details look at 'DATA DIAGNOSIS' in 'Check CytoSense-Diagnosis-Meta Data' 1.29.3: Maximum FL Yellow HS: suspect = 3 1.29.13 Solution - Particle rate too high? 1.29.23 Solution - Trigger too sensitive? 1.29.33 Solution - Details look at 'DATA DIAGNOSIS' in 'Check CytoSense-Diagnosis-Meta Data' 1.29.4: Maximum FL Orange HS: suspect = 5 1.29.14 Solution - Particle rate too high? 1.29.24 Solution - Trigger too sensitive? 1.29.34 Solution - Details look at 'DATA DIAGNOSIS' in 'Check CytoSense-Diagnosis-Meta Data' 1.29.5: Maximum FL Red HS: suspect = 3 1.29.15 Solution - Particle rate too high? 1.29.25 Solution - Trigger too sensitive? 1.29.35 Solution - Details look at 'DATA DIAGNOSIS' in 'Check CytoSense-Diagnosis-Meta Data'