EasyClus check & Info of last file ----- Processed at : 21-Jul-2023 16:46:01 ----- 1. The cytometric analysis is performed on machine IFREMER 2. The CytoUSB software used is CytoUSB Version: 5.11.1.25 3. The filename is FLR26 2023-04-19 08h05.cyz 4. The analysis is performed on 19-04-2023 time 08:05:48 5. The number of measured particles is 95909 6. The total concentration found (e.g. only SWS trigg.) is 27 particles/microliter 7. The concentration found (all used triggers) is 27 particles/microliter 8. The preconcentration (before analysis) found (all used triggers) is 45 particles/microliter 9. The flush time found (between samples) is 60 seconds 10. The effective analysis volume is 3512.2826 microliter 11. The pumped volume is 5299.2 microliter 12. The particle-rate is 301 particles passing the laser beam/second 13. The used sample flow rate is 11.0405 microliter/second 14. The used block size memory is 4 kB 15. Trigger 1: FL Red at threshold level 26 mV 16. Trigger 2: not used at threshold level -999 17. Temperature sensor PMT is : 29.5 C 18. Temperature sensor Sheath is : 28.8 C 19. Temperature sensor Sheath is : 28.0 C 20. Temperature sensor System is : 26.0 C 21. Pressure sensor difference is : -271 mBar 22. Pressure sensor absolute is : 255 mBar 23. Sample pump 1: Last calibration of the sample pump was : 02-Févr-2023 24. Alignment check 1 by curvature detector is 0.72951 (1=perfect) 25. Alignment check 2 by curvature detector is 20% (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 0 (0% of 95909 measured particles) 28. FWS: Maximum signal height found is 2909 mV 29. FWS: Nr of particles with signal height <10 mV is high (174 part. (0 %)) 30. SWS HS: Nr of saturated signals is 826 (0% of 95909 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 (120 part. (0 %)) 33. FL Red HS: Nr of saturated signals is 0 (0% of 95909 measured particles) 34. FL Red HS: Maximum signal height found is 4569 mV 35. FL Red HS: Nr of particles with signal height <10 mV is high (4 part. (0 %)) 36. FL Yellow HS: Nr of saturated signals is 0 (0% of 95909 measured particles) 37. FL Yellow HS: Maximum signal height found is 70 mV 38. FL Yellow HS: Nr of particles with signal height <10 mV is high (95886 part. (100 %)) 39. FL Orange HS: Nr of saturated signals is 0 (0% of 95909 measured particles) 40. FL Orange HS: Maximum signal height found is 2587 mV 41. FL Orange HS: Nr of particles with signal height <10 mV is high (82171 part. (86 %)) 42. FL Green HS: Nr of saturated signals is 0 (0% of 95909 measured particles) 43. FL Green HS: Maximum signal height found is 958 mV 44. FL Green HS: Nr of particles with signal height <10 mV is high (95767 part. (100 %)) 45. FWS: Nr of block shaped signals is 433 (0.45147%) 46. SWS HS: Nr of block shaped signals is 403 (0.42019%) 47. FL Red HS: Nr of block shaped signals is 18 (0.018768%) 48. FL Orange HS: Nr of block shaped signals is 11 (0.011469%) 49. FL Green HS: Nr of block shaped signals is 7 (0.0072986%) 50. Suspect nr: 0=low, 10=high 51. Maximum FWS: suspect = 0 52. Maximum SWS HS: suspect = 0 53. Maximum FL Red HS: suspect = 0 54. Maximum FL Yellow HS: suspect = 0 55. Maximum FL Orange HS: suspect = 2 56. Maximum FL Green HS: suspect = 2 57. SWS: PMT level is 60 58. FL Red: PMT level is 80 59. FL Yellow: PMT level is 90 60. FL Orange: PMT level is 94 61. FL Green: PMT level is 94 -------------------------- 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 1.05 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. (Only older CytoSenses) 1.09.1 Solution - Increase block size memory if the total particle rate is high enough (but prevent time outs) OR set to 'auto' 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 1.23 Warning: FWS-L and -R might indicate an air bubble problem 20% (100%=perfect) 1.23.1 Solution - Pinch valve to get rid of the air bubble or blockage by something different 1.26 Warning: FL Yellow HS: Biggest particles found is only 70 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 (86 %) 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 Green 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)