----- Processed at : 10-Jul-2024 15:47:35 ----- 1. The cytometric analysis is performed on machine RWS Systeem 2 2. The CytoBuoy instrument serial-nr is CS-2019-92 3. The CytoUSB software used is CytoUSB Version: 5.8.3.31 4. The filename is CS_2019_92_prot_2 2022-01-27 14h31.cyz 5. The analysis is performed on 27-01-2022 time 14:31:36 6. The number of measured particles is 4587 7. The total concentration found (e.g. only SWS trigg.) is 1562 particles/microliter 8. The concentration found (all used triggers) is 8 particles/microliter 9. The preconcentration (before analysis) found (all used triggers) is 8 particles/microliter 10. The flush time found (between samples) is 130 seconds 11. The effective analysis volume is 567.7529 microliter 12. The pumped volume is 573 microliter 13. The particle-rate is 2990 particles passing the laser beam/second 14. The used sample flow rate is 1.9134 microliter/second 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 : 32.2 C 18. Temperature sensor Sheath is : 31.7 C 19. Temperature sensor Laser is : 30.9 C 20. Temperature sensor System is : 26.7 C 21. Pressure sensor difference is : -226 mBar 22. Pressure sensor absolute is : 177 mBar 23. Sample pump 1: Last calibration of the sample pump was : 16-Feb-2021 24. Alignment check 1 by curvature detector is 0.73385 (1=perfect) 25. Alignment check 2 by curvature detector is 29.6% (>70%=perfect) 26. Saturated signals (flattened peaks) occur when the dynamic range of the (HS)detector is reached 27. FWS: Nr of saturated signals is 1 (0% of 4587 measured particles) 28. FWS: Maximum signal height found is 14848 mV 29. FWS: Nr of particles with signal height <10 mV is high (1 part. (0 %)) 30. SWS HS: Nr of saturated signals is 6 (0% of 4587 measured particles) 31. SWS HS: Maximum signal height found is 7488 mV 32. SWS HS: Nr of particles with signal height <10 mV is high (0 part. (0 %)) 33. FL Yellow HS: Nr of saturated signals is 1 (0% of 4587 measured particles) 34. FL Yellow HS: Maximum signal height found is 7360 mV 35. FL Yellow HS: Nr of particles with signal height <10 mV is high (2904 part. (63 %)) 36. FL Orange HS: Nr of saturated signals is 1 (0% of 4587 measured particles) 37. FL Orange HS: Maximum signal height found is 7360 mV 38. FL Orange HS: Nr of particles with signal height <10 mV is high (3623 part. (79 %)) 39. FL Red HS: Nr of saturated signals is 1 (0% of 4587 measured particles) 40. FL Red HS: Maximum signal height found is 6496 mV 41. FL Red HS: Nr of particles with signal height <10 mV is high (1350 part. (29 %)) 42. SWS HS: Nr of block shaped signals is 1 (0.021801%) 43. Suspect nr: 0=low, 10=high 44. Maximum FWS: suspect = 0 45. Maximum SWS HS: suspect = 1 46. Maximum SWS LS: suspect = 2 47. Maximum FL Yellow HS: suspect = 0 48. Maximum FL Yellow LS: suspect = 3 49. Maximum FL Orange HS: suspect = 3 50. Maximum FL Red HS: suspect = 2 51. Maximum FL Red LS: suspect = 2 52. SWS HS: PMT level is 56 53. SWS LS: PMT level is 46 54. FL Yellow HS: PMT level is 85 55. FL Yellow LS: PMT level is 68 56. Fl Orange: PMT level is 79 57. FL Red HS: PMT level is 110 58. FL Red LS: PMT level is 91 59. IIF: Imaging in Flow pictures available. Nr of images is 886 -------------------------- 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) 1.07 Warning: Analysis with high particle rate (2990), meaning you have higher chance of coincidence 1.07.1 Solution - Decrease, use lower sample rate 1.07.2 Solution - Is threshold in noise level? Increase threshold and find out of particle rate is suddenly lower and quite stable 1.07.3 Solution - Is the sheath fluid clean enough? 1.21 Warning: Last calibration of sample pump towards analysis date was more than 90 days. (16-Feb-2021) 1.21.1 Solution - Check if the pumped analysis volume is the real volume by weighing 1.23 Warning: Percentage ratio FWS-L/-R data within bandwidth is small which might indicate an alignment problem 29.6% (>70%=perfect) 1.23.1 Solution - Maybe an air bubble or blockage problem? Pinch valve to get rid of the air bubble or blockage by something different 1.24 Warning: SWS LS: Maximum is 247 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 48 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 24 and too low. 1.24.1 Solution - Put LS PMT lower. Ratio HS/LS should be around 100. 1.27 Warning: FL Yellow HS : Nr of particles with signal height <10 mV is high (63 %) 1.27.1 Solution - Increase, set higher trigger threshold (to prevent measuring noise) 1.27.2 Solution - Decrease, set lower particle sample rate. Maybe caused by an overload of particles/ noise in your sample 1.27.3 Solution - Clean sheath, clean cuvette, clean tubing? (noise can be introduced by dirty walls) 1.27 Warning: FL Orange HS : Nr of particles with signal height <10 mV is high (79 %) 1.27.1 Solution - Increase, set higher trigger threshold (to prevent measuring noise) 1.27.2 Solution - Decrease, set lower particle sample rate. Maybe caused by an overload of particles/ noise in your sample 1.27.3 Solution - Clean sheath, clean cuvette, clean tubing? (noise can be introduced by dirty walls) 1.25 Warning: Suspected particles data blocks found! 1.29.1: Maximum FL Yellow LS: suspect = 3 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 FL Orange HS: suspect = 3 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'