EasyClus check & Info of last file ----- Processed at : 29-Nov-2022 15:25:18 ----- 1. The cytometric analysis is performed on machine Cefas 2. The CytoUSB software used is CytoUSB Version: 5.7.1.0 3. The filename is nano_cend18_22 2022-11-29 15u15.cyz 4. The analysis is performed on 29-11-2022 time 15:15:04 5. The number of measured particles is 33162 6. The total concentration found (e.g. only SWS trigg.) is 9 particles/microliter 7. The concentration found (all used triggers) is 9 particles/microliter 8. The preconcentration (before analysis) found (all used triggers) is -1 particles/microliter 9. The flush time found (between samples) is 240 seconds 10. The effective analysis volume is 3580.4782 microliter 11. The pumped volume is 5022 microliter 12. The particle-rate is 77 particles passing the laser beam/second 13. The used sample flow rate is 8.3654 microliter/second 14. The used block size memory is 4 kB 15. Trigger 1: FL Red at threshold level 21 mV 16. Trigger 2: not used at threshold level -999 17. Temperature sensor PMT is : -999.0 C 18. Temperature sensor Sheath is : -999.0 C 19. Temperature sensor Sheath is : 23.1 C 20. Temperature sensor System is : -999.0 C 21. Pressure sensor difference is : -999 mBar 22. Pressure sensor absolute is : -999 mBar 23. Sample pump 1: Last calibration of the sample pump was : 01-Jan-0001 24. Alignment check 1 by curvature detector is 1.4298 (1=perfect) 25. Alignment check 2 by curvature detector is 52% (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 1 (0% of 33162 measured particles) 28. FWS: Maximum signal height found is 7980 mV 29. FWS: Nr of particles with signal height <10 mV is high (5293 part. (16 %)) 30. SWS HS: Nr of saturated signals is 0 (0% of 33162 measured particles) 31. SWS HS: Maximum signal height found is 4251 mV 32. SWS HS: Nr of particles with signal height <10 mV is high (22851 part. (69 %)) 33. FL Yellow HS: Nr of saturated signals is 0 (0% of 33162 measured particles) 34. FL Yellow HS: Maximum signal height found is 4407 mV 35. FL Yellow HS: Nr of particles with signal height <10 mV is high (23932 part. (72 %)) 36. FL Orange HS: Nr of saturated signals is 0 (0% of 33162 measured particles) 37. FL Orange HS: Maximum signal height found is 4327 mV 38. FL Orange HS: Nr of particles with signal height <10 mV is high (22102 part. (67 %)) 39. FL Red HS: Nr of saturated signals is 0 (0% of 33162 measured particles) 40. FL Red HS: Maximum signal height found is 4289 mV 41. FL Red HS: Nr of particles with signal height <10 mV is high (35 part. (0 %)) 42. FWS: Nr of block shaped signals is 213 (0.6423%) 43. SWS HS: Nr of block shaped signals is 181 (0.54581%) 44. FL Yellow HS: Nr of block shaped signals is 159 (0.47946%) 45. FL Red HS: Nr of block shaped signals is 29 (0.087449%) 46. FL Orange HS: Nr of block shaped signals is 1 (0.0030155%) 47. Suspect nr: 0=low, 10=high 48. Maximum FWS: suspect = 0 49. Maximum SWS HS: suspect = 0 50. Maximum FL Yellow HS: suspect = 0 51. Maximum FL Orange HS: suspect = 0 52. Maximum FL Red HS: suspect = 0 53. SWS HS: PMT level is 0 54. FL Yellow HS: PMT level is 0 55. FL Orange HS: PMT level is 0 56. FL Red HS: PMT level is 0 -------------------------- 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' 1.16 Warning: Pressure difference is high. 1.16.1 Solution - Check if there is something blocking from sample inlet to sheath (filter block, tubing) 1.17 Warning: Inner absolute pressure in fluidics towards outside is high. 1.17.1 Solution - Check if there is something blocking in the fluidics pathways (filter block, cuvette, tubing) 1.21 Warning: Last calibration of sample pump towards analysis date was more than 90 days. (01-Jan-0001) 1.21.1 Solution - Check if the pumped analysis volume is the real volume by weighing 1.22 Warning: FWS-L and -R detector signals are quite different (ratio 1.4298(1=perfect)) and might indicate an alignment problem 1.22.1 Solution - Probably you have an air bubble in your system. Pinch valve to get rid of it 1.22.2 Solution - Improve alignment of FWS-L and -R detector towards sample core 1.23 Warning: FWS-L and -R might indicate an air bubble problem 52% (100%=perfect) 1.23.1 Solution - Pinch valve to get rid of the air bubble or blockage by something different 1.27 Warning: SWS HS : Nr of particles with signal height <10 mV is high (69 %) 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 Yellow HS : Nr of particles with signal height <10 mV is high (72 %) 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 (67 %) 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)