EasyClus check & Info of last file ----- Processed at : 03-Aug-2022 14:10:01 ----- 1. The cytometric analysis is performed on machine ULCO II 2. The CytoUSB software used is CytoUSB Version: 5.11.1.6 3. The filename is Pico_MAREL 2022-08-03 13h59.cyz 4. The analysis is performed on 03-08-2022 time 14:01:58 5. The number of measured particles is 7313 6. The total concentration found (e.g. only SWS trigg.) is 2151 particles/microliter 7. The concentration found (all used triggers) is 18 particles/microliter 8. The preconcentration (before analysis) found (all used triggers) is 66 particles/microliter 9. The flush time found (between samples) is 131 seconds 10. The effective analysis volume is 399.7116 microliter 11. The pumped volume is 684.48 microliter 12. The particle-rate is 4004 particles passing the laser beam/second 13. The used sample flow rate is 1.8614 microliter/second 14. The used block size memory is 64 kB 15. Trigger 1: SWS at threshold level 35 mV 16. Trigger 2: Total FWS at threshold level 12 17. Trigger 3: Maximum FL Red at threshold level 8 18. Temperature sensor PMT is : 32.3 C 19. Temperature sensor Sheath is : 29.9 C 20. Temperature sensor Sheath is : 34.1 C 21. Temperature sensor System is : 33.4 C 22. Pressure sensor difference is : -268 mBar 23. Pressure sensor absolute is : 194 mBar 24. Sample pump 1: Last calibration of the sample pump was : 10-Janv-2022 25. Alignment check 1 by curvature detector is 0.8379 (1=perfect) 26. Alignment check 2 by curvature detector is 37% (100%=perfect) 27. Saturated signals (flattened peaks) occur when the dynamic range of the (HS)detector is reached 28. FWS: Nr of saturated signals is 1 (0% of 7313 measured particles) 29. FWS: Maximum signal height found is 6847 mV 30. FWS: Nr of particles with signal height <10 mV is high (550 part. (8 %)) 31. SWS HS: Nr of saturated signals is 1560 (0% of 7313 measured particles) 32. SWS HS: Maximum signal height found is 5000 mV 33. SWS HS: Nr of particles with signal height <10 mV is high (4 part. (0 %)) 34. FL Yellow HS: Nr of saturated signals is 0 (0% of 7313 measured particles) 35. FL Yellow HS: Maximum signal height found is 1267 mV 36. FL Yellow HS: Nr of particles with signal height <10 mV is high (7207 part. (99 %)) 37. FL Orange HS: Nr of saturated signals is 5 (0% of 7313 measured particles) 38. FL Orange HS: Maximum signal height found is 5000 mV 39. FL Orange HS: Nr of particles with signal height <10 mV is high (6749 part. (92 %)) 40. FL Red HS: Nr of saturated signals is 7 (0% of 7313 measured particles) 41. FL Red HS: Maximum signal height found is 5000 mV 42. FL Red HS: Nr of particles with signal height <10 mV is high (1566 part. (21 %)) 43. FL Orange HS: Nr of block shaped signals is 2 (0.027349%) 44. Suspect nr: 0=low, 10=high 45. Maximum FWS: suspect = 0 46. Maximum SWS HS: suspect = 0 47. Maximum FL Yellow HS: suspect = 0 48. Maximum FL Orange HS: suspect = 0 49. Maximum FL Red HS: suspect = 2 50. SWS: PMT level is 65 51. FL Yellow: PMT level is 100 52. FL Orange: PMT level is 95 53. FL Red: PMT level is 95 -------------------------- 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=64k (default=4k) e.g. to 128k 1.07 Warning: Analysis with high particle rate (4004), 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? 1.11 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. (10-Janv-2022) 1.21.1 Solution - Check if the pumped analysis volume is the real volume by weighing 1.23 Warning: FWS-L and -R might indicate an air bubble problem 37% (100%=perfect) 1.23.1 Solution - Pinch valve to get rid of the air bubble or blockage by something different 1.24 Warning: SWS HS: Quite a lot of particles (21% have signal heights of 1.24.1 Solution - 5000 mV or higher. Check if your PMT is not too high! 1.25 Warning: SWS HS: Relative large amount of particles is saturated in (0) 1.25.1 Solution - Decrease, set lower SWS HS PMT voltage in CytoUSB (make less sensitive) 1.27 Warning: FL Yellow HS : Nr of particles with signal height <10 mV is high (99 %) 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 (92 %) 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)