EIJ_SWS15_FLRed6_auto_phyto_600s 2020-11-23 08h01.cyz ----- Processed at : 23-Nov-2020 08:12:46 ----- 1. The cytometric analysis is performed on machine Eijsden 2. The CytoUSB software used is CytoUSB Version: 5.8.3.42 3. The filename is EIJ_SWS15_FLRed6_auto_phyto_600s 2020-11-23 08h01.cyz 4. The analysis is performed on 23-11-2020 time 08:02:31 5. The number of measured particles is 82 6. The total concentration found (e.g. only SWS trigg.) is 2 particles/microliter 7. The concentration found (all used triggers) is 0 particles/microliter 8. The preconcentration (before analysis) found (all used triggers) is 0 particles/microliter 9. The flush time found (between samples) is 50 seconds 10. The effective analysis volume is 2407.6604 microliter 11. The pumped volume is 2724 microliter 12. The particle-rate is 10 particles passing the laser beam/second 13. The used sample flow rate is 4.5388 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 : 29 C 18. Temperature sensor Sheath is : 25 C 19. Temperature sensor Sheath is : 29 C 20. Pressure sensor difference is : -345 mBar 21. Pressure sensor absolute is : 319 mBar 22. Sample pump 1: Last calibration of the sample pump was : 02-Jul-2020 23. Alignment check 1 by curvature detector is 1.2559 (1=perfect) 24. Alignment check 2 by curvature detector is 95% (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 (1% of 82 measured particles) 27. FWS: Maximum signal height found is 9823 mV 28. FWS: Nr of particles with signal height <10 mV is high (3 part. (4 %)) 29. SWS HS: Nr of saturated signals is 5 (6% of 82 measured particles) 30. SWS HS: Maximum signal height found is 5000 mV 31. SWS HS: Nr of particles with signal height <10 mV is high (0 part. (0 %)) 32. FL Yellow HS: Nr of saturated signals is 0 (0% of 82 measured particles) 33. FL Yellow HS: Maximum signal height found is 573 mV 34. FL Yellow HS: Nr of particles with signal height <10 mV is high (35 part. (43 %)) 35. FL Orange HS: Nr of saturated signals is 0 (0% of 82 measured particles) 36. FL Orange HS: Maximum signal height found is 331 mV 37. FL Orange HS: Nr of particles with signal height <10 mV is high (54 part. (66 %)) 38. FL Red HS: Nr of saturated signals is 0 (0% of 82 measured particles) 39. FL Red HS: Maximum signal height found is 38 mV 40. FL Red HS: Nr of particles with signal height <10 mV is high (75 part. (91 %)) 41. FL Yellow HS: Nr of block shaped signals is 9 (10.9756%) 42. Suspect nr: 0=low, 10=high 43. Maximum FWS: suspect = 0 44. Maximum SWS HS: suspect = 0 45. Maximum FL Yellow HS: suspect = 0 46. Maximum FL Orange HS: suspect = 0 47. Maximum FL Red HS: suspect = 0 48. Maximum SWS LS: suspect = 0 49. Maximum FL Yellow LS: suspect = 0 50. Maximum FL Red LS: suspect = 0 51. SWS HS: PMT level is 53 52. FL Yellow HS: PMT level is 78 53. FL Orange HS: PMT level is 77 54. FL Red HS: PMT level is 97 55. SWS LS: PMT level is 25 56. FL Yellow LS: PMT level is 62 57. FL Red LS: PMT level is 70 -------------------------- 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 1.04 Warning: Your flush time (50 seconds) is quite small 1.04.1 Solution - Flush longer e.g. 80 sec. 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) 1.17 Warning: Last calibration of sample pump towards analysis date was more than 90 days. (02-Jul-2020) 1.17.1 Solution - Check if the pumped analysis volume is the real volume by weighing 1.20 Warning: FL Yellow LS: HS-LS ratio is 1 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 2 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 1 and too low. 1.20.1 Solution - Put LS PMT lower. Ratio HS/LS should be around 100. 1.22 Warning: FL Orange HS: Biggest particles found is only 331 mV. Detector sensitivity might be too low. 1.22.1 Solution - Increase, set higher FL Orange HS PMT voltage in CytoUSB (make more sensitive) 1.23 Warning: FL Orange HS : Nr of particles with signal height <10 mV is high (66 %) 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) 1.22 Warning: FL Red HS: Biggest particles found is only 38 mV. Detector sensitivity might be too low. 1.22.1 Solution - Increase, set higher FL Red HS PMT voltage in CytoUSB (make more sensitive) 1.23 Warning: FL Red HS : Nr of particles with signal height <10 mV is high (91 %) 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 (11%) 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)