EasyClus check & Info of last file ----- Processed at : 03-May-2021 22:07:03 ----- 1. The cytometric analysis is performed on machine ULCO II 2. The CytoUSB software used is CytoUSB Version: 5.8.3.41 3. The filename is MAREL_Pico_flr12_5ul_5min 2021-05-03 21h59.cyz 4. The analysis is performed on 03-05-2021 time 22:01:21 5. The number of measured particles is 27684 6. The total concentration found (e.g. only SWS trigg.) is 31 particles/microliter 7. The concentration found (all used triggers) is 31 particles/microliter 8. The preconcentration (before analysis) found (all used triggers) is 827 particles/microliter 9. The flush time found (between samples) is 50 seconds 10. The effective analysis volume is 891.1667 microliter 11. The pumped volume is 1386 microliter 12. The particle-rate is 144 particles passing the laser beam/second 13. The used sample flow rate is 4.62 microliter/second 14. The used block size memory is 4 kB 15. Trigger 1: FL Red at threshold level 12 mV 16. Trigger 2: not used at threshold level -999 17. Temperature sensor PMT is : 24.6 C 18. Temperature sensor Sheath is : 22.2 C 19. Temperature sensor Sheath is : 26.5 C 20. Temperature sensor System is : 25.5 C 21. Pressure sensor difference is : -374 mBar 22. Pressure sensor absolute is : 314 mBar 23. Sample pump 1: Last calibration of the sample pump was : 01-Jan-1990 24. Alignment check 1 by curvature detector is 1.0318 (1=perfect) 25. Alignment check 2 by curvature detector is 73% (100%=perfect) 26. Saturated signals (flattened peaks) occur when the dynamic range of the (HS)detector is reached 27. FWS: Nr of saturated signals is 12 (0% of 27684 measured particles) 28. FWS: Maximum signal height found is 10000 mV 29. FWS: Nr of particles with signal height <10 mV is high (647 part. (2 %)) 30. SWS HS: Nr of saturated signals is 5206 (19% of 27684 measured particles) 31. SWS HS: Maximum signal height found is 5000 mV 32. SWS HS: Nr of particles with signal height <10 mV is high (55 part. (0 %)) 33. FL Yellow HS: Nr of saturated signals is 0 (0% of 27684 measured particles) 34. FL Yellow HS: Maximum signal height found is 185 mV 35. FL Yellow HS: Nr of particles with signal height <10 mV is high (27653 part. (100 %)) 36. FL Orange HS: Nr of saturated signals is 0 (0% of 27684 measured particles) 37. FL Orange HS: Maximum signal height found is 514 mV 38. FL Orange HS: Nr of particles with signal height <10 mV is high (27345 part. (99 %)) 39. FL Red HS: Nr of saturated signals is 3 (0% of 27684 measured particles) 40. FL Red HS: Maximum signal height found is 5000 mV 41. FL Red HS: Nr of particles with signal height <10 mV is high (2084 part. (8 %)) 42. SWS HS: Nr of block shaped signals is 2411 (8.709%) 43. FWS: Nr of block shaped signals is 181 (0.65381%) 44. Suspect nr: 0=low, 10=high 45. Maximum FWS: suspect = 0 46. Maximum SWS HS: suspect = 0 47. Maximum FL Yellow HS: suspect = 2 48. Maximum FL Orange HS: suspect = 1 49. Maximum FL Red HS: suspect = 0 50. SWS: PMT level is 60 51. FL Yellow: PMT level is 90 52. FL Orange: PMT level is 85 53. FL Red: PMT level is 85 -------------------------- Warnings info -------------------------- 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 1.04 Warning: Your flush time (50 seconds) is quite small 1.04.1 Solution - Flush longer e.g. 80 sec. 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) 1.21 Warning: Last calibration of sample pump towards analysis date was more than 90 days. (01-Jan-1990) 1.21.1 Solution - Check if the pumped analysis volume is the real volume by weighing 1.24 Warning: SWS HS: Quite a lot of particles (19% 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 (19) 1.25.1 Solution - Decrease, set lower SWS HS PMT voltage in CytoUSB (make less sensitive) 1.26 Warning: FL Yellow HS: Biggest particles found is only 185 mV. Detector sensitivity might be too low. 1.26.1 Solution - Increase, set higher FL Yellow HS PMT voltage in CytoUSB (make more sensitive) 1.27 Warning: FL Yellow HS : Nr of particles with signal height <10 mV is high (100 %) 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 (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) 1.28 Warning: SWS HS: A relative large amount of particles are (rather) block shaped (9%) 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 SWS HS PMT voltage in CytoUSB (make less sensitive)