EasyClus check & Info of last file ----- Processed at : 26-Nov-2020 00:15:55 ----- 1. The cytometric analysis is performed on machine Markermeer 2. The CytoUSB software used is CytoUSB Version: 5.8.4.8 3. The filename is CS2019-91 MW prot2 2020-11-26 00h03.cyz 4. The analysis is performed on 26-11-2020 time 00:03:27 5. The number of measured particles is 23152 6. The total concentration found (e.g. only SWS trigg.) is 955 particles/microliter 7. The concentration found (all used triggers) is 72 particles/microliter 8. The preconcentration (before analysis) found (all used triggers) is 72 particles/microliter 9. The flush time found (between samples) is 50 seconds 10. The effective analysis volume is 319.4225 microliter 11. The pumped volume is 321 microliter 12. The particle-rate is 1026 particles passing the laser beam/second 13. The used sample flow rate is 1.0746 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 : 128 C 18. Temperature sensor Sheath is : 9 C 19. Temperature sensor Sheath is : 9 C 20. Pressure sensor difference is : -288 mBar 21. Pressure sensor absolute is : 340 mBar 22. Sample pump 1: Last calibration of the sample pump was : 17-Feb-2020 23. Alignment check 1 by curvature detector is 0.99551 (1=perfect) 24. Alignment check 2 by curvature detector is 99% (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 5 (0% of 23152 measured particles) 27. FWS: Maximum signal height found is 14720 mV 28. FWS: Nr of particles with signal height <10 mV is high (47 part. (0 %)) 29. SWS HS: Nr of saturated signals is 1 (0% of 23152 measured particles) 30. SWS HS: Maximum signal height found is 5824 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 1 (0% of 23152 measured particles) 33. FL Yellow HS: Maximum signal height found is 5600 mV 34. FL Yellow HS: Nr of particles with signal height <10 mV is high (20630 part. (89 %)) 35. FL Orange HS: Nr of saturated signals is 1 (0% of 23152 measured particles) 36. FL Orange HS: Maximum signal height found is 5312 mV 37. FL Orange HS: Nr of particles with signal height <10 mV is high (12564 part. (54 %)) 38. FL Red HS: Nr of saturated signals is 6 (0% of 23152 measured particles) 39. FL Red HS: Maximum signal height found is 7360 mV 40. FL Red HS: Nr of particles with signal height <10 mV is high (5811 part. (25 %)) 41. SWS HS: Nr of block shaped signals is 20 (0.086386%) 42. FWS: Nr of block shaped signals is 13 (0.056151%) 43. FL Red HS: Nr of block shaped signals is 12 (0.051831%) 44. FL Orange HS: Nr of block shaped signals is 4 (0.017277%) 45. FL Yellow HS: Nr of block shaped signals is 2 (0.0086386%) 46. Suspect nr: 0=low, 10=high 47. Maximum FWS: suspect = 0 48. Maximum SWS HS: suspect = 0 49. Maximum SWS LS: suspect = 0 50. Maximum FL Yellow HS: suspect = 3 51. Maximum FL Yellow LS: suspect = 0 52. Maximum FL Orange HS: suspect = 0 53. Maximum FL Red HS: suspect = 0 54. Maximum FL Red LS: suspect = 0 55. SWS HS: PMT level is 51 56. SWS LS: PMT level is 46 57. FL Yellow HS: PMT level is 87 58. FL Yellow LS: PMT level is 70 59. Fl Orange: PMT level is 96 60. FL Red HS: PMT level is 118 61. FL Red LS: PMT level is 80 62. IIF: Imaging in Flow pictures available. Nr of images is 251 -------------------------- Warnings info -------------------------- 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) 1.12 Warning: Temperature sensor PMT is high. Inner temperature instrument is high 1.12.1 Solution - Shut laser off if not used, airco in instrument room 1.17 Warning: Last calibration of sample pump towards analysis date was more than 90 days. (17-Feb-2020) 1.17.1 Solution - Check if the pumped analysis volume is the real volume by weighing 1.20 Warning: SWS LS: Maximum is 336 mV if HS is around 4500 mV and too high. 1.20.1 Solution - Put LS PMT lower. Maximum LS around 50 mV (if HS is around 4500 mV). 1.20 Warning: SWS LS: HS-LS ratio is 49 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 128 and too high. 1.20.1 Solution - Put LS PMT higher. Ratio HS/LS should be around 100. 1.20 Warning: FL Red LS: Maximum is 109 mV if HS is around 4500 mV and too high. 1.20.1 Solution - Put LS PMT lower. Maximum LS around 50 mV (if HS is around 4500 mV). 1.20 Warning: FL Red LS: HS-LS ratio is 73 and too low. 1.20.1 Solution - Put LS PMT lower. Ratio HS/LS should be around 100. 1.23 Warning: FL Yellow HS : Nr of particles with signal height <10 mV is high (89 %) 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) 1.23 Warning: FL Orange HS : Nr of particles with signal height <10 mV is high (54 %) 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) 1.25 Warning: Suspected particles data blocks found! 1.25.1: Maximum FL Yellow HS: suspect = 3 1.25.1 Solution - Particle rate too high? 1.25.1 Solution - Trigger too sensitive? 1.25.1 Solution - Details look at 'DATA DIAGNOSIS' in 'Check CytoSense-Diagnosis-Meta Data'