19-Nov-21 20:06:54 Trimming A by 90 90, reference by 85 90 Trimming A by 90 95, reference by 85 90 Trimming A by 100 100, reference by 85 90 Trimming A by 105 105, reference by 85 90 Rough eucentricity: changing Z by -7.98 to -25.68, finished. 105071.047: (Initial) alignment to map 29 (50) 105071.047: Target X, Y: 215.338 -317.595 105071.047: Map matches low dose View; staying in low dose but with exp 0.405 intensity 0.19459 bin 4 Trimming A by 23 100 20 105, reference by 115 164 95 20 Disparity in stage position after aligning to center of map frame: 0.05 0.24 Aligning to target with image shift equivalent to stage shift: 0.06 0.24 105078.515: Map matches low dose Preview; staying in low dose but with exp 0.202 intensity 0.12072 bin 4 105094.000: Realigned to map item itself with image shift of -10.5 2.4 pixels Disparity in stage position after aligning to map item itself: 0.07 0.29 (align shift 0.01 0.00) Finished aligning to item in 2 rounds Opened new file W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070.mrc STARTING TILT SERIES WITH AUTOMATIC TERMINATION AFTER ERRORS. 105102.047: Skipping reverse tilt: dir -1 start 0.00 revang 3.00 revtilt 2.99 maxtilt 69.90 maxerr 0.10 Measured defocus = -9.58 microns changed by 9.48 to target Measured defocus = 0.23 microns changed by -0.33 to target 105137.047: SetIntensity called with factor 0.9962 105137.047: Changing (ideal) Record exposure by 0.9962 to 1.5918 105137.047: Frame time for constraint is 0.1598 105137.047: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105137.047: Skipping reverse tilt: dir -1 start -10.00 revang -7.00 revtilt 2.99 maxtilt 69.90 maxerr 0.10 Need regular tracking - Need autofocus Tracking image shifted -0.006, -0.024 um; position -0.041, -0.029 Measured defocus = -0.00 microns changed by -0.10 to target Tracking image shifted -0.000, -0.001 um; position -0.041, -0.028 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0000_-10.0.tif Frame alignment results: distance raw = 6.5 smoothed = 5.0 Weighted resid mean = 0.05 max max = 0.07 Mean unweighted max = 0.25 Saved Z = 0 at 0.00 min Tilt = -10.00 Mean = 45 Exp = 1.592 X/Y/Z = -0.041 -0.028 4.156 105157.094: Saved state 0 at -10.0: def 4.056 IS 0.048 -0.003 int 0.12073 exp 1.592 105157.094: SetIntensity called with factor 1.0002 105157.094: Changing (ideal) Record exposure by 1.0002 to 1.5920 105157.109: Frame time for constraint is 0.1598 105157.109: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Need regular tracking - Need autofocus Tracking image shifted -0.005, 0.037 um; position -0.035, -0.065 Measured defocus = -0.17 microns changed by 0.07 to target Tracking image shifted 0.000, -0.002 um; position -0.035, -0.063 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0001_-12.0.tif Frame alignment results: distance raw = 3.8 smoothed = 2.4 Weighted resid mean = 0.05 max max = 0.08 Mean unweighted max = 0.12 Record image shifted -0.002, 0.004 um Tracking reference shifted by 0.8,-3.1 pixels, -0.002,0.004 um Saved Z = 1 at 0.48 min Tilt = -11.99 Mean = 46 Exp = 1.592 X/Y/Z = -0.034 -0.067 4.231 105186.047: Saved state 1 at -12.0: def 4.131 IS 0.060 -0.041 int 0.12073 exp 1.592 105186.047: SetIntensity called with factor 1.0005 105186.047: Changing (ideal) Record exposure by 1.0005 to 1.5927 105186.047: Frame time for constraint is 0.1598 105186.062: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Predicted X = -0.027 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Y = -0.105 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Z = 4.305 StdErr = 0.000 nFit = 2 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.008, -0.036 um; position -0.035, -0.069 Measured defocus = -0.06 microns changed by -0.04 to target Tracking image shifted -0.000, -0.002 um; position -0.035, -0.067 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0002_-14.0.tif Frame alignment results: distance raw = 2.6 smoothed = 1.4 Weighted resid mean = 0.05 max max = 0.07 Mean unweighted max = 0.11 Record image shifted -0.002, 0.006 um Tracking reference shifted by 0.5,-4.7 pixels, -0.002,0.006 um Saved Z = 2 at 0.97 min Tilt = -14.00 Mean = 46 Exp = 1.593 X/Y/Z = -0.033 -0.073 4.269 105215.156: Saved state 2 at -14.0: def 4.169 IS 0.063 -0.047 int 0.12073 exp 1.593 105215.234: Restored state 0 at -10.0 for tiltInd 3 def 4.056 IS 0.048 -0.003 105215.234: SetIntensity called with factor 1.0002 105215.234: Changing (ideal) Record exposure by 1.0002 to 1.5920 105215.234: Frame time for constraint is 0.1598 105215.234: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105219.140: recentlyReversed: dir 1 ang -7.998520 revtilt -5.000887 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.006 0.032 um, 0.5 2.3 % Error in Z Prediction -0.036 Need regular tracking - Need autofocus Tracking image shifted 0.081, -0.148 um; position -0.122, 0.120 Measured defocus = -0.03 microns changed by -0.07 to target Tracking image shifted 0.001, -0.005 um; position -0.122, 0.124 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0003_-8.0.tif Frame alignment results: distance raw = 4.6 smoothed = 2.6 Weighted resid mean = 0.05 max max = 0.09 Mean unweighted max = 0.16 Record image shifted -0.002, -0.008 um Tracking reference shifted by 2.3,5.2 pixels, -0.002,-0.008 um Saved Z = 3 at 1.50 min Tilt = -8.00 Mean = 46 Exp = 1.592 X/Y/Z = -0.121 0.132 4.084 105246.890: Saved state 3 at -8.0: def 3.984 IS 0.040 0.181 int 0.12073 exp 1.592 105246.890: SetIntensity called with factor 1.0005 105246.906: Changing (ideal) Record exposure by 1.0005 to 1.5927 105246.906: Frame time for constraint is 0.1598 105246.906: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105250.406: recentlyReversed: dir 1 ang -5.999265 revtilt -3.004632 back 3.000000 3mte 0.300000 Need regular tracking - Need autofocus Tracking image shifted 0.013, -0.046 um; position -0.134, 0.177 Measured defocus = -0.05 microns changed by -0.05 to target Tracking image shifted 0.000, -0.004 um; position -0.134, 0.181 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0004_-6.0.tif Frame alignment results: distance raw = 4.0 smoothed = 2.6 Weighted resid mean = 0.04 max max = 0.07 Mean unweighted max = 0.27 Record image shifted 0.001, -0.006 um Tracking reference shifted by 0.3,4.4 pixels, 0.001,-0.006 um Saved Z = 4 at 2.02 min Tilt = -6.00 Mean = 45 Exp = 1.593 X/Y/Z = -0.135 0.187 4.032 105278.094: Saved state 4 at -6.0: def 3.932 IS 0.026 0.238 int 0.12073 exp 1.593 105278.172: Restored state 2 at -14.0 for tiltInd 5 def 4.169 IS 0.063 -0.047 105278.187: SetIntensity called with factor 1.0008 105278.187: Changing (ideal) Record exposure by 1.0008 to 1.5940 105278.187: Frame time for constraint is 0.1598 105278.187: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Predicted X = -0.028 StdErr = 0.005 nFit = 3 nDrop = 0 Predicted Y = -0.101 StdErr = 0.024 nFit = 3 nDrop = 0 Predicted Z = 4.332 StdErr = 0.027 nFit = 3 nDrop = 0 Need autofocus Measured defocus = -0.19 microns changed by 0.09 to target 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0005_-16.0.tif Frame alignment results: distance raw = 2.6 smoothed = 1.5 Weighted resid mean = 0.04 max max = 0.06 Mean unweighted max = 0.13 Record image shifted 0.077, -0.078 um Saved Z = 5 at 2.49 min Tilt = -16.00 Mean = 45 Exp = 1.594 X/Y/Z = -0.106 -0.023 4.420 105306.609: Saved state 5 at -16.0: def 4.320 IS 0.101 0.037 int 0.12073 exp 1.594 105306.609: SetIntensity called with factor 1.0011 105306.609: Changing (ideal) Record exposure by 1.0011 to 1.5957 105306.609: Frame time for constraint is 0.1598 105306.625: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.077 0.078 um, 5.6 5.6 % Error in Z Prediction 0.089 Predicted X = -0.129 StdErr = 0.055 nFit = 3 nDrop = 0 Predicted Y = -0.011 StdErr = 0.042 nFit = 3 nDrop = 0 Predicted Z = 4.496 StdErr = 0.084 nFit = 3 nDrop = 0 Need regular tracking - Tracking image shifted -0.015, -0.053 um; position -0.115, 0.042 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0006_-18.0.tif Frame alignment results: distance raw = 2.6 smoothed = 1.7 Weighted resid mean = 0.04 max max = 0.08 Mean unweighted max = 0.13 Record image shifted -0.006, 0.017 um Tracking reference shifted by 2.0,-12.9 pixels, -0.006,0.017 um Saved Z = 6 at 2.89 min Tilt = -17.99 Mean = 46 Exp = 1.596 X/Y/Z = -0.109 0.025 4.496 105330.344: Saved state 6 at -18.0: def 4.396 IS 0.081 0.081 int 0.12073 exp 1.596 105330.422: Restored state 4 at -6.0 for tiltInd 7 def 3.932 IS 0.026 0.238 105330.422: SetIntensity called with factor 1.0008 105330.437: Changing (ideal) Record exposure by 1.0008 to 1.5940 105330.437: Frame time for constraint is 0.1598 105330.437: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105335.203: recentlyReversed: dir 1 ang -3.999010 revtilt -1.010376 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.020 0.036 um, 1.5 2.6 % Predicted X = -0.149 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Y = 0.243 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Z = 3.981 StdErr = 0.000 nFit = 2 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.014, 0.033 um; position -0.163, 0.209 Measured defocus = -0.02 microns changed by -0.08 to target Tracking image shifted 0.001, -0.004 um; position -0.164, 0.214 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0007_-4.0.tif Frame alignment results: distance raw = 4.7 smoothed = 3.5 Weighted resid mean = 0.04 max max = 0.07 Mean unweighted max = 0.17 Record image shifted -0.001, -0.006 um Tracking reference shifted by 1.8,4.4 pixels, -0.001,-0.006 um Saved Z = 7 at 3.43 min Tilt = -4.00 Mean = 45 Exp = 1.594 X/Y/Z = -0.163 0.220 3.897 105363.031: Saved state 7 at -4.0: def 3.797 IS 0.035 0.282 int 0.12073 exp 1.594 105363.047: SetIntensity called with factor 1.0011 105363.047: Changing (ideal) Record exposure by 1.0011 to 1.5957 105363.047: Frame time for constraint is 0.1598 105363.062: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105366.453: recentlyReversed: dir 1 ang -1.999255 revtilt 0.997878 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.014 -0.023 um, 1.0 1.6 % Error in Z Prediction -0.084 Predicted X = -0.182 StdErr = 0.010 nFit = 3 nDrop = 0 Predicted Y = 0.268 StdErr = 0.017 nFit = 3 nDrop = 0 Predicted Z = 3.818 StdErr = 0.062 nFit = 3 nDrop = 0 Need regular tracking - Tracking image shifted -0.017, 0.077 um; position -0.165, 0.190 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0008_-2.0.tif Frame alignment results: distance raw = 2.6 smoothed = 1.3 Weighted resid mean = 0.04 max max = 0.07 Mean unweighted max = 0.24 Record image shifted 0.001, -0.009 um Tracking reference shifted by 0.4,6.7 pixels, 0.001,-0.009 um Saved Z = 8 at 3.86 min Tilt = -2.00 Mean = 45 Exp = 1.596 X/Y/Z = -0.166 0.199 3.818 105388.719: Saved state 8 at -2.0: def 3.718 IS 0.047 0.266 int 0.12073 exp 1.596 105388.797: Restored state 6 at -18.0 for tiltInd 9 def 4.396 IS 0.081 0.081 105388.797: SetIntensity called with factor 1.0014 105388.797: Changing (ideal) Record exposure by 1.0014 to 1.5979 105388.812: Frame time for constraint is 0.1598 105388.812: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.016 -0.068 um, 1.1 4.9 % Predicted X = -0.158 StdErr = 0.052 nFit = 3 nDrop = 0 Predicted Y = 0.075 StdErr = 0.001 nFit = 3 nDrop = 0 Predicted Z = 4.591 StdErr = 0.111 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.020, 0.035 um; position -0.139, 0.040 Measured defocus = -0.03 microns changed by -0.07 to target Tracking image shifted -0.000, -0.002 um; position -0.138, 0.042 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0009_-20.0.tif Frame alignment results: distance raw = 4.8 smoothed = 3.2 Weighted resid mean = 0.05 max max = 0.08 Mean unweighted max = 0.13 Record image shifted -0.003, 0.017 um Tracking reference shifted by 0.1,-12.9 pixels, -0.003,0.017 um Saved Z = 9 at 4.38 min Tilt = -19.99 Mean = 46 Exp = 1.598 X/Y/Z = -0.135 0.025 4.524 105419.969: Saved state 9 at -20.0: def 4.424 IS 0.104 0.095 int 0.12073 exp 1.598 105419.969: SetIntensity called with factor 1.0017 105419.984: Changing (ideal) Record exposure by 1.0017 to 1.6006 105419.984: Frame time for constraint is 0.1598 105419.984: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.023 -0.050 um, 1.7 3.6 % Error in Z Prediction -0.067 Predicted X = -0.146 StdErr = 0.017 nFit = 3 nDrop = 0 Predicted Y = 0.057 StdErr = 0.037 nFit = 3 nDrop = 0 Predicted Z = 4.618 StdErr = 0.089 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.016, 0.060 um; position -0.130, -0.003 Measured defocus = -0.05 microns changed by -0.05 to target Tracking image shifted -0.000, -0.000 um; position -0.130, -0.003 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0010_-22.0.tif Frame alignment results: distance raw = 4.1 smoothed = 3.5 Weighted resid mean = 0.05 max max = 0.08 Mean unweighted max = 0.17 Record image shifted -0.002, 0.010 um Tracking reference shifted by 0.1,-7.3 pixels, -0.002,0.010 um Saved Z = 10 at 4.90 min Tilt = -21.99 Mean = 46 Exp = 1.601 X/Y/Z = -0.128 -0.012 4.571 105451.265: Saved state 10 at -22.0: def 4.471 IS 0.115 0.058 int 0.12073 exp 1.601 105451.344: Restored state 8 at -2.0 for tiltInd 11 def 3.718 IS 0.047 0.266 105451.344: SetIntensity called with factor 1.0014 105451.359: Changing (ideal) Record exposure by 1.0014 to 1.5979 105451.359: Frame time for constraint is 0.1598 105451.359: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105456.453: recentlyReversed: dir 1 ang 0.002500 revtilt 2.994133 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.018 -0.069 um, 1.3 5.0 % Error in Z Prediction -0.047 Predicted X = -0.186 StdErr = 0.019 nFit = 3 nDrop = 0 Predicted Y = 0.214 StdErr = 0.040 nFit = 3 nDrop = 0 Predicted Z = 3.724 StdErr = 0.083 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.007, 0.031 um; position -0.194, 0.184 Measured defocus = -0.22 microns changed by 0.12 to target Tracking image shifted 0.001, -0.006 um; position -0.194, 0.189 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0011_0.0.tif Frame alignment results: distance raw = 4.0 smoothed = 3.3 Weighted resid mean = 0.07 max max = 0.12 Mean unweighted max = 0.22 Record image shifted -0.001, -0.002 um Tracking reference shifted by 1.2,0.9 pixels, -0.001,-0.002 um Saved Z = 11 at 5.45 min Tilt = 0.00 Mean = 45 Exp = 1.598 X/Y/Z = -0.193 0.191 3.845 105484.344: Saved state 11 at 0.0: def 3.745 IS 0.074 0.272 int 0.12073 exp 1.598 105484.344: SetIntensity called with factor 1.0017 105484.359: Changing (ideal) Record exposure by 1.0017 to 1.6006 105484.359: Frame time for constraint is 0.1598 105484.359: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105487.765: recentlyReversed: dir 1 ang 1.999755 revtilt 4.997888 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.007 -0.024 um, 0.5 1.7 % Error in Z Prediction 0.121 Predicted X = -0.204 StdErr = 0.017 nFit = 3 nDrop = 0 Predicted Y = 0.174 StdErr = 0.009 nFit = 3 nDrop = 0 Predicted Z = 3.773 StdErr = 0.099 nFit = 3 nDrop = 0 Need regular tracking - Tracking image shifted -0.009, -0.030 um; position -0.196, 0.204 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0012_2.0.tif Frame alignment results: distance raw = 2.7 smoothed = 1.4 Weighted resid mean = 0.05 max max = 0.07 Mean unweighted max = 0.15 Record image shifted 0.001, -0.008 um Tracking reference shifted by 0.6,6.1 pixels, 0.001,-0.008 um Saved Z = 12 at 5.88 min Tilt = 2.00 Mean = 45 Exp = 1.601 X/Y/Z = -0.196 0.212 3.773 105510.062: Saved state 12 at 2.0: def 3.673 IS 0.067 0.293 int 0.12073 exp 1.601 105510.140: Restored state 10 at -22.0 for tiltInd 13 def 4.471 IS 0.115 0.058 105510.140: SetIntensity called with factor 1.0020 105510.140: Changing (ideal) Record exposure by 1.0020 to 1.6038 105510.140: Frame time for constraint is 0.1598 105510.156: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.008 0.038 um, 0.6 2.7 % Predicted X = -0.129 StdErr = 0.037 nFit = 4 nDrop = 0 Predicted Y = -0.095 StdErr = 0.008 nFit = 4 nDrop = 0 Predicted Z = 4.658 StdErr = 0.062 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.021, -0.045 um; position -0.151, -0.051 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0013_-24.0.tif Frame alignment results: distance raw = 3.0 smoothed = 2.3 Weighted resid mean = 0.06 max max = 0.08 Mean unweighted max = 0.20 Record image shifted -0.003, 0.011 um Tracking reference shifted by 1.0,-8.2 pixels, -0.003,0.011 um Saved Z = 13 at 6.33 min Tilt = -23.99 Mean = 46 Exp = 1.604 X/Y/Z = -0.147 -0.061 4.658 105536.703: Saved state 13 at -24.0: def 4.558 IS 0.155 0.025 int 0.12073 exp 1.604 105536.719: SetIntensity called with factor 1.0024 105536.719: Changing (ideal) Record exposure by 1.0024 to 1.6076 105536.734: Frame time for constraint is 0.1598 105536.734: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.018 0.034 um, 1.3 2.5 % Predicted X = -0.148 StdErr = 0.040 nFit = 4 nDrop = 0 Predicted Y = -0.159 StdErr = 0.020 nFit = 5 nDrop = 0 Predicted Z = 4.729 StdErr = 0.072 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.008, -0.056 um; position -0.140, -0.104 Measured defocus = -0.18 microns changed by 0.08 to target Tracking image shifted 0.000, -0.001 um; position -0.140, -0.103 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0014_-26.0.tif Frame alignment results: distance raw = 9.5 smoothed = 7.9 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.23 Record image shifted -0.002, 0.012 um Tracking reference shifted by -0.0,-8.7 pixels, -0.002,0.012 um Saved Z = 14 at 6.81 min Tilt = -25.99 Mean = 45 Exp = 1.608 X/Y/Z = -0.138 -0.115 4.805 105565.875: Saved state 14 at -26.0: def 4.705 IS 0.172 -0.027 int 0.12073 exp 1.608 105565.969: Restored state 12 at 2.0 for tiltInd 15 def 3.673 IS 0.067 0.293 105565.969: SetIntensity called with factor 1.0020 105565.969: Changing (ideal) Record exposure by 1.0020 to 1.6038 105565.984: Frame time for constraint is 0.1598 105565.984: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105571.953: recentlyReversed: dir 1 ang 3.999010 revtilt 6.993643 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.010 0.045 um, 0.8 3.2 % Error in Z Prediction 0.076 Predicted X = -0.215 StdErr = 0.017 nFit = 3 nDrop = 0 Predicted Y = 0.213 StdErr = 0.023 nFit = 3 nDrop = 0 Predicted Z = 3.716 StdErr = 0.120 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.009, -0.030 um; position -0.224, 0.244 Measured defocus = -0.03 microns changed by -0.07 to target Tracking image shifted -0.000, -0.006 um; position -0.224, 0.250 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0015_4.0.tif Frame alignment results: distance raw = 5.5 smoothed = 4.4 Weighted resid mean = 0.05 max max = 0.10 Mean unweighted max = 0.31 Record image shifted -0.000, 0.001 um Tracking reference shifted by 0.2,-0.9 pixels, -0.000,0.001 um Saved Z = 15 at 7.38 min Tilt = 4.00 Mean = 45 Exp = 1.604 X/Y/Z = -0.224 0.249 3.642 105599.828: Saved state 15 at 4.0: def 3.542 IS 0.073 0.340 int 0.12073 exp 1.604 105599.844: SetIntensity called with factor 1.0023 105599.844: Changing (ideal) Record exposure by 1.0023 to 1.6076 105599.859: Frame time for constraint is 0.1598 105599.859: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105603.265: recentlyReversed: dir 1 ang 5.999765 revtilt 8.998398 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.009 0.035 um, 0.6 2.6 % Error in Z Prediction -0.074 Predicted X = -0.240 StdErr = 0.032 nFit = 4 nDrop = 0 Predicted Y = 0.312 StdErr = 0.010 nFit = 4 nDrop = 0 Predicted Z = 3.603 StdErr = 0.098 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.013, 0.038 um; position -0.227, 0.274 Measured defocus = -0.02 microns changed by -0.08 to target Tracking image shifted 0.000, -0.005 um; position -0.227, 0.279 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0016_6.0.tif Frame alignment results: distance raw = 4.7 smoothed = 3.9 Weighted resid mean = 0.07 max max = 0.10 Mean unweighted max = 0.18 Record image shifted 0.000, -0.004 um Tracking reference shifted by 0.2,2.7 pixels, 0.000,-0.004 um Saved Z = 16 at 7.90 min Tilt = 6.00 Mean = 45 Exp = 1.608 X/Y/Z = -0.227 0.283 3.527 105630.937: Saved state 16 at 6.0: def 3.427 IS 0.060 0.373 int 0.12073 exp 1.608 105631.031: Restored state 14 at -26.0 for tiltInd 17 def 4.705 IS 0.172 -0.027 105631.031: SetIntensity called with factor 1.0027 105631.031: Changing (ideal) Record exposure by 1.0027 to 1.6119 105631.047: Frame time for constraint is 0.1598 105631.047: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.012 -0.029 um, 0.9 2.1 % Error in Z Prediction -0.076 Predicted X = -0.141 StdErr = 0.037 nFit = 4 nDrop = 0 Predicted Y = -0.196 StdErr = 0.017 nFit = 5 nDrop = 1 Predicted Z = 4.859 StdErr = 0.061 nFit = 5 nDrop = 0 Need regular tracking - Tracking image shifted 0.024, -0.090 um; position -0.165, -0.106 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0017_-28.0.tif Frame alignment results: distance raw = 4.2 smoothed = 3.3 Weighted resid mean = 0.05 max max = 0.08 Mean unweighted max = 0.24 Record image shifted -0.003, 0.012 um Tracking reference shifted by 0.8,-9.4 pixels, -0.003,0.012 um Saved Z = 17 at 8.34 min Tilt = -27.99 Mean = 45 Exp = 1.612 X/Y/Z = -0.162 -0.118 4.859 105657.734: Saved state 17 at -28.0: def 4.759 IS 0.194 -0.018 int 0.12073 exp 1.612 105657.734: SetIntensity called with factor 1.0030 105657.750: Changing (ideal) Record exposure by 1.0030 to 1.6168 105657.750: Frame time for constraint is 0.1598 105657.750: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.020 0.077 um, 1.5 5.6 % Predicted X = -0.172 StdErr = 0.041 nFit = 4 nDrop = 0 Predicted Y = -0.175 StdErr = 0.041 nFit = 6 nDrop = 0 Predicted Z = 4.941 StdErr = 0.067 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.009, -0.107 um; position -0.163, -0.068 Measured defocus = -0.16 microns changed by 0.06 to target Tracking image shifted 0.000, -0.001 um; position -0.163, -0.067 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0018_-30.0.tif Frame alignment results: distance raw = 6.5 smoothed = 5.7 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.23 Record image shifted -0.002, 0.012 um Tracking reference shifted by 0.0,-9.1 pixels, -0.002,0.012 um Saved Z = 18 at 8.82 min Tilt = -29.99 Mean = 45 Exp = 1.617 X/Y/Z = -0.161 -0.079 5.002 105686.547: Saved state 18 at -30.0: def 4.902 IS 0.175 0.017 int 0.12073 exp 1.617 105686.640: Restored state 16 at 6.0 for tiltInd 19 def 3.427 IS 0.060 0.373 105686.640: SetIntensity called with factor 1.0027 105686.656: Changing (ideal) Record exposure by 1.0027 to 1.6119 105686.656: Frame time for constraint is 0.1598 105686.672: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105693.094: recentlyReversed: dir 1 ang 7.999520 revtilt 10.998153 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.011 0.096 um, 0.8 6.9 % Error in Z Prediction 0.061 Predicted X = -0.243 StdErr = 0.032 nFit = 4 nDrop = 0 Predicted Y = 0.345 StdErr = 0.017 nFit = 5 nDrop = 0 Predicted Z = 3.483 StdErr = 0.078 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.006, 0.059 um; position -0.249, 0.286 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0019_8.0.tif Frame alignment results: distance raw = 3.1 smoothed = 3.0 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.27 Record image shifted -0.000, -0.008 um Tracking reference shifted by 1.4,5.9 pixels, -0.000,-0.008 um Saved Z = 19 at 9.31 min Tilt = 8.00 Mean = 45 Exp = 1.612 X/Y/Z = -0.249 0.295 3.483 105715.500: Saved state 19 at 8.0: def 3.383 IS 0.073 0.394 int 0.12073 exp 1.612 105715.515: SetIntensity called with factor 1.0030 105715.531: Changing (ideal) Record exposure by 1.0030 to 1.6167 105715.531: Frame time for constraint is 0.1598 105715.547: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105718.937: recentlyReversed: dir 1 ang 9.999275 revtilt 12.997908 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.006 -0.051 um, 0.4 3.7 % Predicted X = -0.257 StdErr = 0.028 nFit = 4 nDrop = 0 Predicted Y = 0.318 StdErr = 0.020 nFit = 5 nDrop = 1 Predicted Z = 3.407 StdErr = 0.088 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.011, 0.036 um; position -0.247, 0.283 Measured defocus = -0.03 microns changed by -0.07 to target Tracking image shifted -0.000, -0.005 um; position -0.247, 0.288 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0020_10.0.tif Frame alignment results: distance raw = 3.9 smoothed = 2.8 Weighted resid mean = 0.09 max max = 0.14 Mean unweighted max = 0.18 Record image shifted 0.000, 0.001 um Tracking reference shifted by -0.3,-0.4 pixels, 0.000,0.001 um Saved Z = 20 at 9.82 min Tilt = 10.00 Mean = 45 Exp = 1.617 X/Y/Z = -0.247 0.287 3.337 105746.547: Saved state 20 at 10.0: def 3.237 IS 0.074 0.387 int 0.12073 exp 1.617 105746.625: Restored state 18 at -30.0 for tiltInd 21 def 4.902 IS 0.175 0.017 105746.640: SetIntensity called with factor 1.0034 105746.640: Changing (ideal) Record exposure by 1.0034 to 1.6223 105746.656: Frame time for constraint is 0.1598 105746.656: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.010 -0.031 um, 0.8 2.3 % Error in Z Prediction -0.071 Predicted X = -0.179 StdErr = 0.038 nFit = 4 nDrop = 0 Predicted Y = -0.021 StdErr = 0.028 nFit = 5 nDrop = 1 Predicted Z = 5.098 StdErr = 0.040 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.006, 0.061 um; position -0.186, -0.082 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0021_-32.0.tif Frame alignment results: distance raw = 3.8 smoothed = 2.8 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.19 Record image shifted -0.003, 0.016 um Tracking reference shifted by -0.1,-11.8 pixels, -0.003,0.016 um Saved Z = 21 at 10.28 min Tilt = -31.99 Mean = 45 Exp = 1.622 X/Y/Z = -0.183 -0.098 5.098 105773.812: Saved state 21 at -32.0: def 4.998 IS 0.202 0.012 int 0.12073 exp 1.622 105773.828: SetIntensity called with factor 1.0037 105773.828: Changing (ideal) Record exposure by 1.0037 to 1.6283 105773.844: Frame time for constraint is 0.1598 105773.844: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.004 -0.077 um, 0.3 5.6 % Predicted X = -0.192 StdErr = 0.031 nFit = 4 nDrop = 0 Predicted Y = -0.042 StdErr = 0.043 nFit = 6 nDrop = 0 Predicted Z = 5.197 StdErr = 0.045 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.016, 0.073 um; position -0.176, -0.115 Measured defocus = 0.01 microns changed by -0.11 to target Tracking image shifted -0.001, 0.004 um; position -0.175, -0.119 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0022_-34.0.tif Frame alignment results: distance raw = 6.6 smoothed = 5.8 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.26 Record image shifted -0.002, 0.012 um Tracking reference shifted by -0.1,-8.7 pixels, -0.002,0.012 um Saved Z = 22 at 10.77 min Tilt = -33.99 Mean = 45 Exp = 1.628 X/Y/Z = -0.173 -0.131 5.087 105803.062: Saved state 22 at -34.0: def 4.987 IS 0.210 -0.023 int 0.12073 exp 1.628 105803.140: Restored state 20 at 10.0 for tiltInd 23 def 3.237 IS 0.074 0.387 105803.140: SetIntensity called with factor 1.0034 105803.156: Changing (ideal) Record exposure by 1.0034 to 1.6221 105803.172: Frame time for constraint is 0.1598 105803.172: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105809.922: recentlyReversed: dir 1 ang 11.999030 revtilt 14.992663 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.019 -0.089 um, 1.4 6.4 % Error in Z Prediction -0.110 Predicted X = -0.260 StdErr = 0.018 nFit = 3 nDrop = 0 Predicted Y = 0.269 StdErr = 0.010 nFit = 5 nDrop = 1 Predicted Z = 3.230 StdErr = 0.011 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.004, 0.011 um; position -0.264, 0.258 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0023_12.0.tif Frame alignment results: distance raw = 4.0 smoothed = 3.8 Weighted resid mean = 0.09 max max = 0.15 Mean unweighted max = 0.23 Record image shifted -0.001, -0.008 um Tracking reference shifted by 1.8,5.3 pixels, -0.001,-0.008 um Saved Z = 23 at 11.26 min Tilt = 12.00 Mean = 44 Exp = 1.622 X/Y/Z = -0.263 0.265 3.230 105832.969: Saved state 23 at 12.0: def 3.130 IS 0.098 0.376 int 0.12073 exp 1.622 105832.984: SetIntensity called with factor 1.0037 105832.984: Changing (ideal) Record exposure by 1.0037 to 1.6282 105833.000: Frame time for constraint is 0.1598 105833.015: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105836.297: recentlyReversed: dir 1 ang 13.999285 revtilt 16.997918 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.003 -0.003 um, 0.2 0.3 % Predicted X = -0.266 StdErr = 0.028 nFit = 4 nDrop = 0 Predicted Y = 0.221 StdErr = 0.004 nFit = 5 nDrop = 1 Predicted Z = 3.128 StdErr = 0.013 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.006, -0.045 um; position -0.260, 0.266 Measured defocus = -0.14 microns changed by 0.04 to target Tracking image shifted 0.001, -0.007 um; position -0.261, 0.273 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0024_14.0.tif Frame alignment results: distance raw = 4.3 smoothed = 1.7 Weighted resid mean = 0.07 max max = 0.12 Mean unweighted max = 0.21 Record image shifted -0.001, 0.002 um Tracking reference shifted by 0.2,-1.2 pixels, -0.001,0.002 um Saved Z = 24 at 11.78 min Tilt = 14.00 Mean = 44 Exp = 1.628 X/Y/Z = -0.260 0.271 3.170 105864.015: Saved state 24 at 14.0: def 3.070 IS 0.093 0.380 int 0.12073 exp 1.628 105864.094: Restored state 22 at -34.0 for tiltInd 25 def 4.987 IS 0.210 -0.023 105864.109: SetIntensity called with factor 1.0041 105864.109: Changing (ideal) Record exposure by 1.0041 to 1.6350 105864.125: Frame time for constraint is 0.1598 105864.140: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.006 0.050 um, 0.4 3.6 % Error in Z Prediction 0.042 Predicted X = -0.184 StdErr = 0.024 nFit = 3 nDrop = 0 Predicted Y = -0.171 StdErr = 0.042 nFit = 5 nDrop = 1 Predicted Z = 5.215 StdErr = 0.077 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.004, 0.013 um; position -0.188, -0.183 Measured defocus = 0.02 microns changed by -0.12 to target Tracking image shifted -0.000, 0.003 um; position -0.188, -0.186 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0025_-36.0.tif Frame alignment results: distance raw = 6.9 smoothed = 5.4 Weighted resid mean = 0.04 max max = 0.08 Mean unweighted max = 0.28 Record image shifted -0.003, 0.015 um Tracking reference shifted by 0.4,-11.2 pixels, -0.003,0.015 um Saved Z = 25 at 12.34 min Tilt = -35.99 Mean = 45 Exp = 1.635 X/Y/Z = -0.184 -0.201 5.091 105897.375: Saved state 25 at -36.0: def 4.991 IS 0.253 -0.078 int 0.12073 exp 1.635 105897.390: SetIntensity called with factor 1.0045 105897.390: Changing (ideal) Record exposure by 1.0045 to 1.6423 105897.406: Frame time for constraint is 0.1598 105897.422: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.000 -0.031 um, 0.0 2.2 % Error in Z Prediction -0.124 Predicted X = -0.177 StdErr = 0.035 nFit = 4 nDrop = 0 Predicted Y = -0.307 StdErr = 0.018 nFit = 5 nDrop = 1 Predicted Z = 5.183 StdErr = 0.070 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted -0.001, -0.071 um; position -0.176, -0.236 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0026_-38.0.tif Frame alignment results: distance raw = 3.4 smoothed = 2.3 Weighted resid mean = 0.08 max max = 0.15 Mean unweighted max = 0.22 Record image shifted -0.003, 0.015 um Tracking reference shifted by 0.0,-11.3 pixels, -0.003,0.015 um Saved Z = 26 at 12.74 min Tilt = -37.99 Mean = 45 Exp = 1.642 X/Y/Z = -0.173 -0.251 5.183 105921.219: Saved state 26 at -38.0: def 5.083 IS 0.267 -0.129 int 0.12073 exp 1.642 105921.312: Restored state 24 at 14.0 for tiltInd 27 def 3.070 IS 0.093 0.380 105921.328: SetIntensity called with factor 1.0041 105921.328: Changing (ideal) Record exposure by 1.0041 to 1.6348 105921.344: Frame time for constraint is 0.1598 105921.344: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105928.969: recentlyReversed: dir 1 ang 15.999040 revtilt 18.999172 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.004 0.056 um, 0.3 4.1 % Predicted X = -0.267 StdErr = 0.025 nFit = 4 nDrop = 0 Predicted Y = 0.251 StdErr = 0.026 nFit = 5 nDrop = 0 Predicted Z = 3.067 StdErr = 0.022 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.017, -0.055 um; position -0.283, 0.307 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0027_16.0.tif Frame alignment results: distance raw = 6.1 smoothed = 5.7 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.30 Record image shifted -0.001, -0.007 um Tracking reference shifted by 1.3,4.7 pixels, -0.001,-0.007 um Saved Z = 27 at 13.24 min Tilt = 16.00 Mean = 44 Exp = 1.635 X/Y/Z = -0.283 0.313 3.067 105951.312: Saved state 27 at 16.0: def 2.967 IS 0.092 0.429 int 0.12073 exp 1.635 105951.328: SetIntensity called with factor 1.0045 105951.344: Changing (ideal) Record exposure by 1.0045 to 1.6421 105951.344: Frame time for constraint is 0.1598 105951.359: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 105955.094: recentlyReversed: dir 1 ang 17.999295 revtilt 21.001427 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.016 0.062 um, 1.2 4.5 % Predicted X = -0.288 StdErr = 0.019 nFit = 3 nDrop = 0 Predicted Y = 0.326 StdErr = 0.036 nFit = 6 nDrop = 0 Predicted Z = 2.974 StdErr = 0.024 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.006, -0.027 um; position -0.283, 0.353 Measured defocus = -0.00 microns changed by -0.10 to target Tracking image shifted -0.000, -0.003 um; position -0.282, 0.356 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0028_18.0.tif Frame alignment results: distance raw = 6.5 smoothed = 5.0 Weighted resid mean = 0.08 max max = 0.14 Mean unweighted max = 0.31 Record image shifted -0.000, -0.001 um Tracking reference shifted by 0.3,0.7 pixels, -0.000,-0.001 um Saved Z = 28 at 13.76 min Tilt = 18.00 Mean = 44 Exp = 1.642 X/Y/Z = -0.282 0.357 2.874 105982.984: Saved state 28 at 18.0: def 2.774 IS 0.071 0.467 int 0.12073 exp 1.642 105983.062: Restored state 26 at -38.0 for tiltInd 29 def 5.083 IS 0.267 -0.129 105983.078: SetIntensity called with factor 1.0049 105983.094: Changing (ideal) Record exposure by 1.0049 to 1.6504 105983.094: Frame time for constraint is 0.1598 105983.109: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.006 0.031 um, 0.5 2.2 % Error in Z Prediction -0.099 Predicted X = -0.177 StdErr = 0.017 nFit = 3 nDrop = 0 Predicted Y = -0.361 StdErr = 0.031 nFit = 6 nDrop = 0 Predicted Z = 5.241 StdErr = 0.079 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.017, -0.103 um; position -0.194, -0.259 Measured defocus = -0.11 microns changed by 0.01 to target Tracking image shifted 0.000, 0.003 um; position -0.194, -0.261 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0029_-40.0.tif Frame alignment results: distance raw = 6.4 smoothed = 5.6 Weighted resid mean = 0.09 max max = 0.12 Mean unweighted max = 0.22 Record image shifted -0.004, 0.017 um Tracking reference shifted by 0.6,-12.8 pixels, -0.004,0.017 um Saved Z = 29 at 14.33 min Tilt = -39.99 Mean = 44 Exp = 1.650 X/Y/Z = -0.190 -0.278 5.247 106016.672: Saved state 29 at -40.0: def 5.147 IS 0.294 -0.143 int 0.12073 exp 1.650 106016.687: SetIntensity called with factor 1.0053 106016.703: Changing (ideal) Record exposure by 1.0053 to 1.6591 106016.703: Frame time for constraint is 0.1598 106016.719: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.013 0.083 um, 1.0 6.0 % Error in Z Prediction 0.006 Predicted X = -0.198 StdErr = 0.034 nFit = 4 nDrop = 0 Predicted Y = -0.320 StdErr = 0.033 nFit = 5 nDrop = 0 Predicted Z = 5.272 StdErr = 0.049 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.006, -0.073 um; position -0.192, -0.247 Measured defocus = -0.10 microns changed by 0.00 to target Tracking image shifted -0.000, 0.003 um; position -0.191, -0.250 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0030_-42.0.tif Frame alignment results: distance raw = 7.1 smoothed = 5.7 Weighted resid mean = 0.08 max max = 0.14 Mean unweighted max = 0.22 Record image shifted -0.003, 0.013 um Tracking reference shifted by 0.1,-10.0 pixels, -0.003,0.013 um Saved Z = 30 at 14.81 min Tilt = -41.99 Mean = 44 Exp = 1.659 X/Y/Z = -0.189 -0.263 5.272 106045.781: Saved state 30 at -42.0: def 5.172 IS 0.286 -0.131 int 0.12073 exp 1.659 106045.859: Restored state 28 at 18.0 for tiltInd 31 def 2.774 IS 0.071 0.467 106045.875: SetIntensity called with factor 1.0049 106045.875: Changing (ideal) Record exposure by 1.0049 to 1.6501 106045.890: Frame time for constraint is 0.1598 106045.890: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 106053.375: recentlyReversed: dir 1 ang 19.999550 revtilt 22.993683 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.009 0.057 um, 0.6 4.1 % Error in Z Prediction -0.000 Predicted X = -0.297 StdErr = 0.017 nFit = 3 nDrop = 0 Predicted Y = 0.438 StdErr = 0.017 nFit = 5 nDrop = 0 Predicted Z = 2.802 StdErr = 0.063 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.010, 0.034 um; position -0.306, 0.404 Measured defocus = -0.11 microns changed by 0.01 to target Tracking image shifted 0.000, -0.004 um; position -0.306, 0.409 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0031_20.0.tif Frame alignment results: distance raw = 6.6 smoothed = 5.5 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.38 Record image shifted -0.001, -0.004 um Tracking reference shifted by 1.2,2.9 pixels, -0.001,-0.004 um Saved Z = 31 at 15.40 min Tilt = 20.00 Mean = 44 Exp = 1.650 X/Y/Z = -0.305 0.413 2.811 106081.078: Saved state 31 at 20.0: def 2.711 IS 0.064 0.529 int 0.12073 exp 1.650 106081.094: SetIntensity called with factor 1.0053 106081.094: Changing (ideal) Record exposure by 1.0053 to 1.6587 106081.109: Frame time for constraint is 0.1598 106081.125: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 106084.375: recentlyReversed: dir 1 ang 21.999305 revtilt 24.998437 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.009 -0.025 um, 0.6 1.8 % Error in Z Prediction 0.009 Predicted X = -0.313 StdErr = 0.018 nFit = 3 nDrop = 0 Predicted Y = 0.489 StdErr = 0.015 nFit = 5 nDrop = 0 Predicted Z = 2.688 StdErr = 0.056 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted -0.015, 0.094 um; position -0.298, 0.395 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0032_22.0.tif Frame alignment results: distance raw = 9.1 smoothed = 7.8 Weighted resid mean = 0.08 max max = 0.12 Mean unweighted max = 0.27 Record image shifted 0.002, -0.006 um Tracking reference shifted by -0.3,4.7 pixels, 0.002,-0.006 um Saved Z = 32 at 15.83 min Tilt = 22.00 Mean = 43 Exp = 1.659 X/Y/Z = -0.299 0.402 2.688 106106.875: Saved state 32 at 22.0: def 2.588 IS 0.064 0.516 int 0.12073 exp 1.659 106106.953: Restored state 30 at -42.0 for tiltInd 33 def 5.172 IS 0.286 -0.131 106106.969: SetIntensity called with factor 1.0057 106106.969: Changing (ideal) Record exposure by 1.0057 to 1.6686 106106.984: Frame time for constraint is 0.1598 106107.000: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.014 -0.087 um, 1.0 6.3 % Predicted X = -0.204 StdErr = 0.031 nFit = 4 nDrop = 0 Predicted Y = -0.231 StdErr = 0.012 nFit = 5 nDrop = 1 Predicted Z = 5.332 StdErr = 0.043 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.006, 0.019 um; position -0.210, -0.250 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0033_-44.0.tif Frame alignment results: distance raw = 3.8 smoothed = 3.0 Weighted resid mean = 0.08 max max = 0.14 Mean unweighted max = 0.22 Record image shifted -0.004, 0.023 um Tracking reference shifted by -0.4,-17.0 pixels, -0.004,0.023 um Saved Z = 33 at 16.30 min Tilt = -43.99 Mean = 44 Exp = 1.669 X/Y/Z = -0.207 -0.273 5.332 106135.406: Saved state 33 at -44.0: def 5.232 IS 0.306 -0.129 int 0.12073 exp 1.669 106135.406: SetIntensity called with factor 1.0062 106135.422: Changing (ideal) Record exposure by 1.0062 to 1.6789 106135.437: Frame time for constraint is 0.1598 106135.437: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.003 -0.042 um, 0.2 3.0 % Predicted X = -0.215 StdErr = 0.025 nFit = 4 nDrop = 0 Predicted Y = -0.231 StdErr = 0.023 nFit = 6 nDrop = 0 Predicted Z = 5.384 StdErr = 0.049 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.006, 0.046 um; position -0.210, -0.277 Measured defocus = -0.07 microns changed by -0.03 to target Tracking image shifted 0.000, 0.005 um; position -0.210, -0.282 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0034_-46.0.tif Frame alignment results: distance raw = 5.7 smoothed = 5.0 Weighted resid mean = 0.07 max max = 0.12 Mean unweighted max = 0.34 Record image shifted -0.003, 0.012 um Tracking reference shifted by 0.8,-9.4 pixels, -0.003,0.012 um Saved Z = 34 at 16.79 min Tilt = -45.98 Mean = 44 Exp = 1.679 X/Y/Z = -0.206 -0.295 5.349 106164.609: Saved state 34 at -46.0: def 5.249 IS 0.316 -0.149 int 0.12073 exp 1.679 106164.719: Restored state 32 at 22.0 for tiltInd 35 def 2.588 IS 0.064 0.516 106164.719: SetIntensity called with factor 1.0057 106164.734: Changing (ideal) Record exposure by 1.0057 to 1.6682 106164.750: Frame time for constraint is 0.1598 106164.765: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 106172.531: recentlyReversed: dir 1 ang 23.999060 revtilt 26.993193 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.009 -0.064 um, 0.7 4.6 % Error in Z Prediction -0.036 Predicted X = -0.313 StdErr = 0.022 nFit = 3 nDrop = 0 Predicted Y = 0.413 StdErr = 0.041 nFit = 5 nDrop = 0 Predicted Z = 2.577 StdErr = 0.062 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.002, 0.053 um; position -0.314, 0.359 Measured defocus = -0.05 microns changed by -0.05 to target Tracking image shifted 0.000, -0.004 um; position -0.314, 0.363 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0035_24.0.tif Frame alignment results: distance raw = 8.4 smoothed = 7.9 Weighted resid mean = 0.09 max max = 0.14 Mean unweighted max = 0.29 Record image shifted -0.003, 0.001 um Tracking reference shifted by 1.8,-1.3 pixels, -0.003,0.001 um Saved Z = 35 at 17.39 min Tilt = 24.00 Mean = 43 Exp = 1.668 X/Y/Z = -0.312 0.361 2.528 106200.312: Saved state 35 at 24.0: def 2.428 IS 0.094 0.487 int 0.12073 exp 1.668 106200.328: SetIntensity called with factor 1.0061 106200.328: Changing (ideal) Record exposure by 1.0061 to 1.6784 106200.344: Frame time for constraint is 0.1598 106200.359: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 106203.562: recentlyReversed: dir 1 ang 25.999315 revtilt 28.998447 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.001 -0.051 um, 0.1 3.7 % Error in Z Prediction -0.049 Predicted X = -0.312 StdErr = 0.014 nFit = 3 nDrop = 0 Predicted Y = 0.294 StdErr = 0.033 nFit = 5 nDrop = 0 Predicted Z = 2.406 StdErr = 0.048 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.007, -0.052 um; position -0.305, 0.346 Measured defocus = -0.14 microns changed by 0.04 to target Tracking image shifted -0.001, -0.004 um; position -0.305, 0.350 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0036_26.0.tif Frame alignment results: distance raw = 11.3 smoothed = 10.6 Weighted resid mean = 0.09 max max = 0.15 Mean unweighted max = 0.36 Record image shifted 0.001, -0.005 um Tracking reference shifted by 0.0,3.6 pixels, 0.001,-0.005 um Saved Z = 36 at 17.91 min Tilt = 26.00 Mean = 43 Exp = 1.678 X/Y/Z = -0.305 0.355 2.449 106231.750: Saved state 36 at 26.0: def 2.349 IS 0.092 0.478 int 0.12073 exp 1.678 106231.828: Restored state 34 at -46.0 for tiltInd 37 def 5.249 IS 0.316 -0.149 106231.844: SetIntensity called with factor 1.0067 106231.859: Changing (ideal) Record exposure by 1.0067 to 1.6901 106231.875: Frame time for constraint is 0.1598 106231.890: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.006 0.060 um, 0.5 4.4 % Error in Z Prediction 0.043 Predicted X = -0.216 StdErr = 0.025 nFit = 4 nDrop = 0 Predicted Y = -0.277 StdErr = 0.036 nFit = 6 nDrop = 0 Predicted Z = 5.409 StdErr = 0.038 nFit = 5 nDrop = 0 Need regular tracking - Tracking image shifted 0.017, 0.059 um; position -0.232, -0.335 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0037_-48.0.tif Frame alignment results: distance raw = 6.6 smoothed = 6.2 Weighted resid mean = 0.08 max max = 0.12 Mean unweighted max = 0.23 Record image shifted -0.006, 0.026 um Tracking reference shifted by 0.5,-19.7 pixels, -0.006,0.026 um Saved Z = 37 at 18.39 min Tilt = -47.98 Mean = 43 Exp = 1.690 X/Y/Z = -0.227 -0.361 5.409 106260.640: Saved state 37 at -48.0: def 5.309 IS 0.365 -0.197 int 0.12073 exp 1.690 106260.656: SetIntensity called with factor 1.0072 106260.672: Changing (ideal) Record exposure by 1.0072 to 1.7022 106260.687: Frame time for constraint is 0.1598 106260.687: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.011 -0.085 um, 0.8 6.1 % Predicted X = -0.239 StdErr = 0.026 nFit = 4 nDrop = 0 Predicted Y = -0.442 StdErr = 0.014 nFit = 5 nDrop = 1 Predicted Z = 5.456 StdErr = 0.041 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.016, -0.038 um; position -0.223, -0.404 Measured defocus = -0.20 microns changed by 0.10 to target Tracking image shifted -0.000, 0.004 um; position -0.222, -0.408 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0038_-50.0.tif Frame alignment results: distance raw = 5.2 smoothed = 4.1 Weighted resid mean = 0.09 max max = 0.14 Mean unweighted max = 0.28 Record image shifted -0.003, 0.018 um Tracking reference shifted by -0.1,-13.7 pixels, -0.003,0.018 um Saved Z = 38 at 18.88 min Tilt = -49.98 Mean = 43 Exp = 1.702 X/Y/Z = -0.219 -0.427 5.554 106289.781: Saved state 38 at -50.0: def 5.454 IS 0.389 -0.259 int 0.12073 exp 1.702 106289.875: Restored state 36 at 26.0 for tiltInd 39 def 2.349 IS 0.092 0.478 106289.890: SetIntensity called with factor 1.0066 106289.890: Changing (ideal) Record exposure by 1.0066 to 1.6895 106289.906: Frame time for constraint is 0.1598 106289.922: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 106298.062: recentlyReversed: dir 1 ang 27.999070 revtilt 30.996203 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.020 0.015 um, 1.5 1.1 % Error in Z Prediction 0.098 Predicted X = -0.308 StdErr = 0.025 nFit = 4 nDrop = 0 Predicted Y = 0.283 StdErr = 0.054 nFit = 5 nDrop = 0 Predicted Z = 2.326 StdErr = 0.047 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.028, -0.052 um; position -0.337, 0.335 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0039_28.0.tif Frame alignment results: distance raw = 17.6 smoothed = 17.0 Weighted resid mean = 0.11 max max = 0.17 Mean unweighted max = 0.31 Record image shifted -0.002, -0.006 um Tracking reference shifted by 2.2,4.1 pixels, -0.002,-0.006 um Saved Z = 39 at 19.40 min Tilt = 28.00 Mean = 42 Exp = 1.689 X/Y/Z = -0.335 0.341 2.326 106320.844: Saved state 39 at 28.0: def 2.226 IS 0.124 0.482 int 0.12073 exp 1.689 106320.859: SetIntensity called with factor 1.0071 106320.859: Changing (ideal) Record exposure by 1.0071 to 1.7015 106320.875: Frame time for constraint is 0.1598 106320.890: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 106324.156: recentlyReversed: dir 1 ang 29.999825 revtilt 32.998457 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.027 0.058 um, 1.9 4.2 % Predicted X = -0.341 StdErr = 0.027 nFit = 3 nDrop = 0 Predicted Y = 0.331 StdErr = 0.024 nFit = 5 nDrop = 0 Predicted Z = 2.212 StdErr = 0.054 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.013, 0.010 um; position -0.328, 0.321 Measured defocus = -0.03 microns changed by -0.07 to target Tracking image shifted -0.000, -0.004 um; position -0.328, 0.325 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0040_30.0.tif Frame alignment results: distance raw = 12.2 smoothed = 11.1 Weighted resid mean = 0.08 max max = 0.13 Mean unweighted max = 0.48 Record image shifted 0.001, 0.001 um Tracking reference shifted by -1.1,-0.8 pixels, 0.001,0.001 um Saved Z = 40 at 19.92 min Tilt = 30.00 Mean = 42 Exp = 1.701 X/Y/Z = -0.329 0.324 2.143 106352.312: Saved state 40 at 30.0: def 2.043 IS 0.127 0.463 int 0.12073 exp 1.701 106352.390: Restored state 38 at -50.0 for tiltInd 41 def 5.454 IS 0.389 -0.259 106352.406: SetIntensity called with factor 1.0077 106352.422: Changing (ideal) Record exposure by 1.0077 to 1.7152 106352.437: Frame time for constraint is 0.1598 106352.453: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.012 -0.007 um, 0.8 0.5 % Error in Z Prediction -0.069 Predicted X = -0.220 StdErr = 0.033 nFit = 4 nDrop = 0 Predicted Y = -0.527 StdErr = 0.012 nFit = 6 nDrop = 0 Predicted Z = 5.582 StdErr = 0.071 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.034, -0.061 um; position -0.253, -0.466 Measured defocus = -0.20 microns changed by 0.10 to target Tracking image shifted -0.000, 0.005 um; position -0.253, -0.471 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0041_-52.0.tif Frame alignment results: distance raw = 6.4 smoothed = 4.5 Weighted resid mean = 0.08 max max = 0.13 Mean unweighted max = 0.40 Record image shifted -0.006, 0.026 um Tracking reference shifted by 0.5,-19.4 pixels, -0.006,0.026 um Saved Z = 41 at 20.50 min Tilt = -51.98 Mean = 42 Exp = 1.715 X/Y/Z = -0.247 -0.496 5.678 106387.234: Saved state 41 at -52.0: def 5.578 IS 0.447 -0.305 int 0.12073 exp 1.715 106387.250: SetIntensity called with factor 1.0082 106387.250: Changing (ideal) Record exposure by 1.0082 to 1.7294 106387.265: Frame time for constraint is 0.1598 106387.281: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.028 0.031 um, 2.0 2.2 % Error in Z Prediction 0.097 Predicted X = -0.252 StdErr = 0.027 nFit = 3 nDrop = 0 Predicted Y = -0.601 StdErr = 0.019 nFit = 6 nDrop = 0 Predicted Z = 5.706 StdErr = 0.073 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.003, -0.109 um; position -0.248, -0.492 Measured defocus = -0.06 microns changed by -0.04 to target Tracking image shifted -0.000, 0.003 um; position -0.248, -0.495 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0042_-54.0.tif Frame alignment results: distance raw = 3.8 smoothed = 2.5 Weighted resid mean = 0.08 max max = 0.13 Mean unweighted max = 0.40 Record image shifted -0.004, 0.021 um Tracking reference shifted by 0.2,-15.8 pixels, -0.004,0.021 um Saved Z = 42 at 21.00 min Tilt = -53.99 Mean = 42 Exp = 1.729 X/Y/Z = -0.244 -0.516 5.664 106416.859: Saved state 42 at -54.0: def 5.564 IS 0.453 -0.324 int 0.12073 exp 1.729 106416.937: Restored state 40 at 30.0 for tiltInd 43 def 2.043 IS 0.127 0.463 106416.953: SetIntensity called with factor 1.0076 106416.969: Changing (ideal) Record exposure by 1.0076 to 1.7144 106416.984: Frame time for constraint is 0.1598 106417.000: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 106425.531: recentlyReversed: dir 1 ang 31.999580 revtilt 34.993213 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.008 0.085 um, 0.5 6.2 % Error in Z Prediction -0.042 Predicted X = -0.347 StdErr = 0.026 nFit = 3 nDrop = 0 Predicted Y = 0.323 StdErr = 0.021 nFit = 5 nDrop = 0 Predicted Z = 2.023 StdErr = 0.041 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.006, 0.011 um; position -0.353, 0.312 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0043_32.0.tif Frame alignment results: distance raw = 13.0 smoothed = 12.0 Weighted resid mean = 0.13 max max = 0.19 Mean unweighted max = 0.32 Record image shifted -0.001, -0.007 um Tracking reference shifted by 1.8,4.6 pixels, -0.001,-0.007 um Saved Z = 43 at 21.52 min Tilt = 32.00 Mean = 41 Exp = 1.714 X/Y/Z = -0.352 0.319 2.023 106448.406: Saved state 43 at 32.0: def 1.923 IS 0.149 0.471 int 0.12073 exp 1.714 106448.406: SetIntensity called with factor 1.0082 106448.422: Changing (ideal) Record exposure by 1.0082 to 1.7284 106448.437: Frame time for constraint is 0.1598 106448.453: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 106451.515: recentlyReversed: dir 1 ang 33.999335 revtilt 36.997967 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.005 -0.004 um, 0.4 0.3 % Predicted X = -0.356 StdErr = 0.021 nFit = 3 nDrop = 0 Predicted Y = 0.305 StdErr = 0.010 nFit = 5 nDrop = 0 Predicted Z = 1.891 StdErr = 0.046 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.013, -0.000 um; position -0.343, 0.306 Measured defocus = -0.17 microns changed by 0.07 to target Tracking image shifted -0.000, -0.002 um; position -0.343, 0.308 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0044_34.0.tif Frame alignment results: distance raw = 14.6 smoothed = 13.4 Weighted resid mean = 0.11 max max = 0.17 Mean unweighted max = 0.43 Record image shifted 0.001, -0.004 um Tracking reference shifted by -0.4,3.0 pixels, 0.001,-0.004 um Saved Z = 44 at 22.04 min Tilt = 34.00 Mean = 41 Exp = 1.728 X/Y/Z = -0.344 0.312 1.956 106479.687: Saved state 44 at 34.0: def 1.856 IS 0.145 0.461 int 0.12073 exp 1.728 106479.765: Restored state 42 at -54.0 for tiltInd 45 def 5.564 IS 0.453 -0.324 106479.781: SetIntensity called with factor 1.0088 106479.797: Changing (ideal) Record exposure by 1.0088 to 1.7446 106479.812: Frame time for constraint is 0.1598 106479.828: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.012 0.007 um, 0.8 0.5 % Error in Z Prediction 0.064 Predicted X = -0.262 StdErr = 0.024 nFit = 3 nDrop = 0 Predicted Y = -0.547 StdErr = 0.029 nFit = 5 nDrop = 0 Predicted Z = 5.773 StdErr = 0.065 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.018, -0.073 um; position -0.280, -0.475 Measured defocus = -0.08 microns changed by -0.02 to target Tracking image shifted -0.001, 0.003 um; position -0.279, -0.478 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0045_-56.0.tif Frame alignment results: distance raw = 7.5 smoothed = 7.2 Weighted resid mean = 0.10 max max = 0.16 Mean unweighted max = 0.36 Record image shifted -0.006, 0.029 um Tracking reference shifted by 0.4,-22.2 pixels, -0.006,0.029 um Saved Z = 45 at 22.64 min Tilt = -55.99 Mean = 41 Exp = 1.745 X/Y/Z = -0.273 -0.507 5.744 106515.390: Saved state 45 at -56.0: def 5.644 IS 0.474 -0.301 int 0.12073 exp 1.745 106515.406: SetIntensity called with factor 1.0095 106515.422: Changing (ideal) Record exposure by 1.0095 to 1.7611 106515.437: Frame time for constraint is 0.1598 106515.453: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.011 0.040 um, 0.8 2.9 % Error in Z Prediction -0.028 Predicted X = -0.281 StdErr = 0.024 nFit = 3 nDrop = 0 Predicted Y = -0.476 StdErr = 0.023 nFit = 5 nDrop = 0 Predicted Z = 5.847 StdErr = 0.066 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.004, 0.014 um; position -0.276, -0.490 Measured defocus = -0.44 microns changed by 0.34 to target Tracking image shifted -0.000, 0.001 um; position -0.276, -0.491 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0046_-58.0.tif Frame alignment results: distance raw = 4.7 smoothed = 3.6 Weighted resid mean = 0.09 max max = 0.14 Mean unweighted max = 0.32 Record image shifted -0.004, 0.027 um Tracking reference shifted by -1.1,-20.5 pixels, -0.004,0.027 um Saved Z = 46 at 23.13 min Tilt = -57.99 Mean = 40 Exp = 1.761 X/Y/Z = -0.273 -0.518 6.187 106545.109: Saved state 46 at -58.0: def 6.087 IS 0.479 -0.311 int 0.12073 exp 1.761 106545.203: Restored state 44 at 34.0 for tiltInd 47 def 1.856 IS 0.145 0.461 106545.219: SetIntensity called with factor 1.0088 106545.234: Changing (ideal) Record exposure by 1.0088 to 1.7436 106545.234: Frame time for constraint is 0.1598 106545.250: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 106553.969: recentlyReversed: dir 1 ang 35.999090 revtilt 38.980724 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.008 -0.042 um, 0.6 3.0 % Error in Z Prediction 0.341 Predicted X = -0.357 StdErr = 0.023 nFit = 3 nDrop = 0 Predicted Y = 0.311 StdErr = 0.006 nFit = 5 nDrop = 0 Predicted Z = 1.821 StdErr = 0.053 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.013, -0.029 um; position -0.370, 0.340 Measured defocus = -0.05 microns changed by -0.05 to target Tracking image shifted -0.001, -0.002 um; position -0.369, 0.342 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0047_36.0.tif Frame alignment results: distance raw = 17.6 smoothed = 16.7 Weighted resid mean = 0.15 max max = 0.25 Mean unweighted max = 0.34 Record image shifted -0.002, -0.005 um Tracking reference shifted by 2.1,3.7 pixels, -0.002,-0.005 um Saved Z = 47 at 23.75 min Tilt = 36.00 Mean = 40 Exp = 1.744 X/Y/Z = -0.367 0.347 1.773 106582.359: Saved state 47 at 36.0: def 1.673 IS 0.148 0.505 int 0.12073 exp 1.744 106582.375: SetIntensity called with factor 1.0094 106582.390: Changing (ideal) Record exposure by 1.0094 to 1.7600 106582.406: Frame time for constraint is 0.1598 106582.422: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 106585.672: recentlyReversed: dir 1 ang 37.999345 revtilt 40.993478 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.010 0.037 um, 0.7 2.7 % Error in Z Prediction -0.047 Predicted X = -0.375 StdErr = 0.041 nFit = 4 nDrop = 0 Predicted Y = 0.381 StdErr = 0.020 nFit = 5 nDrop = 0 Predicted Z = 1.660 StdErr = 0.054 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.015, 0.043 um; position -0.359, 0.338 Measured defocus = -0.23 microns changed by 0.13 to target Tracking image shifted -0.000, -0.002 um; position -0.359, 0.339 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0048_38.0.tif Frame alignment results: distance raw = 17.3 smoothed = 16.7 Weighted resid mean = 0.12 max max = 0.19 Mean unweighted max = 0.50 Record image shifted 0.000, -0.002 um Tracking reference shifted by 0.0,1.3 pixels, 0.000,-0.002 um Saved Z = 48 at 24.28 min Tilt = 38.00 Mean = 39 Exp = 1.760 X/Y/Z = -0.359 0.341 1.788 106614.000: Saved state 48 at 38.0: def 1.688 IS 0.144 0.495 int 0.12073 exp 1.760 106614.094: Restored state 46 at -58.0 for tiltInd 49 def 6.087 IS 0.479 -0.311 106614.109: SetIntensity called with factor 1.0102 106614.125: Changing (ideal) Record exposure by 1.0102 to 1.7790 106614.140: Frame time for constraint is 0.1598 106614.156: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.015 -0.039 um, 1.1 2.8 % Error in Z Prediction 0.128 Predicted X = -0.292 StdErr = 0.022 nFit = 3 nDrop = 0 Predicted Y = -0.478 StdErr = 0.038 nFit = 5 nDrop = 0 Predicted Z = 6.165 StdErr = 0.211 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.011, 0.018 um; position -0.303, -0.497 Measured defocus = 0.21 microns changed by -0.31 to target Tracking image shifted 0.000, 0.004 um; position -0.303, -0.501 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0049_-60.0.tif Frame alignment results: distance raw = 12.5 smoothed = 11.7 Weighted resid mean = 0.13 max max = 0.21 Mean unweighted max = 0.54 Record image shifted -0.007, 0.037 um Tracking reference shifted by -0.1,-27.9 pixels, -0.007,0.037 um Saved Z = 49 at 24.88 min Tilt = -59.99 Mean = 40 Exp = 1.779 X/Y/Z = -0.296 -0.538 5.851 106649.875: Saved state 49 at -60.0: def 5.751 IS 0.508 -0.316 int 0.12073 exp 1.779 106649.890: SetIntensity called with factor 1.0109 106649.906: Changing (ideal) Record exposure by 1.0109 to 1.7984 106649.922: Frame time for constraint is 0.1598 106649.937: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.005 -0.060 um, 0.3 4.3 % Error in Z Prediction -0.314 Predicted X = -0.304 StdErr = 0.018 nFit = 3 nDrop = 0 Predicted Y = -0.552 StdErr = 0.023 nFit = 5 nDrop = 0 Predicted Z = 6.089 StdErr = 0.227 nFit = 6 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.955, 0.648 um; position 0.651, -1.200 Measured defocus = 0.49 microns changed by -0.59 to target Tracking image shifted 0.943, -0.594 um; position -0.292, -0.606 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0050_-62.0.tif Frame alignment results: distance raw = 12.2 smoothed = 11.3 Weighted resid mean = 0.11 max max = 0.19 Mean unweighted max = 0.73 Trimming A by 25 131, reference by 21 21 Record image shifted -0.002, 0.018 um Tracking reference shifted by -0.7,-13.1 pixels, -0.002,0.018 um Saved Z = 50 at 25.38 min Tilt = -61.99 Mean = 33 Exp = 1.798 X/Y/Z = -0.289 -0.624 6.616 106680.140: Saved state 50 at -62.0: def 6.516 IS 0.543 -0.395 int 0.12073 exp 1.798 106680.219: Restored state 48 at 38.0 for tiltInd 51 def 1.688 IS 0.144 0.495 106680.234: SetIntensity called with factor 1.0101 106680.250: Changing (ideal) Record exposure by 1.0101 to 1.7778 106680.265: Frame time for constraint is 0.1598 106680.281: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 106689.250: recentlyReversed: dir 1 ang 39.999100 revtilt 42.997732 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.015 -0.072 um, 1.1 5.2 % Error in Z Prediction 0.527 Predicted X = -0.367 StdErr = 0.041 nFit = 4 nDrop = 0 Predicted Y = 0.368 StdErr = 0.033 nFit = 5 nDrop = 0 Predicted Z = 1.647 StdErr = 0.086 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.021, -0.003 um; position -0.388, 0.371 Measured defocus = -0.08 microns changed by -0.02 to target Tracking image shifted -0.001, -0.003 um; position -0.387, 0.374 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0051_40.0.tif Frame alignment results: distance raw = 22.6 smoothed = 22.0 Weighted resid mean = 0.15 max max = 0.26 Mean unweighted max = 0.59 Record image shifted -0.002, -0.005 um Tracking reference shifted by 2.2,3.1 pixels, -0.002,-0.005 um Saved Z = 51 at 26.00 min Tilt = 40.00 Mean = 39 Exp = 1.778 X/Y/Z = -0.385 0.379 1.621 106717.265: Saved state 51 at 40.0: def 1.521 IS 0.149 0.542 int 0.12073 exp 1.778 106717.281: SetIntensity called with factor 1.0108 106717.297: Changing (ideal) Record exposure by 1.0108 to 1.7970 106717.312: Frame time for constraint is 0.1598 106717.328: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 106720.906: recentlyReversed: dir 1 ang 41.998355 revtilt 44.998987 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.018 0.011 um, 1.3 0.8 % Error in Z Prediction -0.026 Predicted X = -0.397 StdErr = 0.043 nFit = 4 nDrop = 0 Predicted Y = 0.411 StdErr = 0.024 nFit = 6 nDrop = 0 Predicted Z = 1.532 StdErr = 0.068 nFit = 5 nDrop = 0 Need regular tracking - Tracking image shifted -0.022, 0.065 um; position -0.375, 0.345 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0052_42.0.tif Frame alignment results: distance raw = 20.1 smoothed = 19.1 Weighted resid mean = 0.13 max max = 0.23 Mean unweighted max = 0.53 Record image shifted 0.000, -0.001 um Tracking reference shifted by 0.1,1.0 pixels, 0.000,-0.001 um Saved Z = 52 at 26.44 min Tilt = 42.00 Mean = 38 Exp = 1.797 X/Y/Z = -0.375 0.347 1.532 106743.484: Saved state 52 at 42.0: def 1.432 IS 0.155 0.509 int 0.12073 exp 1.797 106743.562: Restored state 50 at -62.0 for tiltInd 53 def 6.516 IS 0.543 -0.395 106743.578: SetIntensity called with factor 1.0117 106743.594: Changing (ideal) Record exposure by 1.0117 to 1.8195 106743.609: Frame time for constraint is 0.1598 106743.625: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.022 -0.064 um, 1.6 4.6 % Predicted X = -0.303 StdErr = 0.023 nFit = 3 nDrop = 0 Predicted Y = -0.713 StdErr = 0.028 nFit = 5 nDrop = 0 Predicted Z = 6.526 StdErr = 0.338 nFit = 6 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.504, 0.243 um; position -0.806, -0.956 Measured defocus = -37.89 microns changed by 16.00 Measured defocus = -20.37 microns changed by 16.00 Measured defocus = -2.97 microns changed by 2.87 to target Measured defocus = -0.11 microns changed by 0.01 to target Redoing Autofocus because measured defocus is too far from prediction Measured defocus = -0.14 microns changed by 0.04 to target Redoing image because of insufficient counts Redoing image because of insufficient counts There have been 3 failures to obtain an image with sufficient counts (2 actual vs. 2 criterion unbinned counts per sec) ONE SIDE OF THE SERIES IS BEING ENDED; GOING ON TO FINISH THE OTHER SIDE BASED ON TILT SERIES POLICY SETTINGS 106784.375: Restored state 52 at 42.0 for tiltInd 55 def 1.432 IS 0.155 0.509 106784.390: SetIntensity called with factor 1.0117 106784.406: Changing (ideal) Record exposure by 1.0117 to 1.8180 106784.437: Frame time for constraint is 0.1598 106784.453: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 106793.531: recentlyReversed: dir 1 ang 43.999110 revtilt 46.997742 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.072 1.060 um, 5.2 76.6 % Need regular tracking - Need autofocus Tracking image shifted 0.021, -0.019 um; position -0.396, 0.366 Measured defocus = 0.45 microns changed by -0.55 to target Tracking image shifted -0.008, 0.001 um; position -0.388, 0.366 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0053_44.0.tif Frame alignment results: distance raw = 29.8 smoothed = 29.0 Weighted resid mean = 0.42 max max = 0.93 Mean unweighted max = 29.40 Record image shifted -0.003, -0.013 um Tracking reference shifted by 3.7,9.0 pixels, -0.003,-0.013 um Saved Z = 53 at 27.70 min Tilt = 44.00 Mean = 37 Exp = 1.818 X/Y/Z = -0.385 0.378 0.987 106819.344: Saved state 53 at 44.0: def 0.887 IS 0.149 0.542 int 0.12073 exp 1.818 106819.359: SetIntensity called with factor 1.0125 106819.375: Changing (ideal) Record exposure by 1.0125 to 1.8408 106819.390: Frame time for constraint is 0.1598 106819.406: Will be constrained to 1.9174 (frame 0.1598, constraint 0.1598) 106822.594: recentlyReversed: dir 1 ang 45.999364 revtilt 48.997997 back 3.000000 3mte 0.300000 Need regular tracking - Need autofocus Tracking image shifted -0.017, 0.023 um; position -0.369, 0.355 Measured defocus = 0.04 microns changed by -0.14 to target Tracking image shifted -0.001, -0.001 um; position -0.368, 0.356 12 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0054_46.0.tif Frame alignment results: distance raw = 28.7 smoothed = 28.2 Weighted resid mean = 0.34 max max = 0.58 Mean unweighted max = 28.17 Record image shifted 0.002, -0.004 um Tracking reference shifted by -0.7,3.1 pixels, 0.002,-0.004 um Saved Z = 54 at 28.23 min Tilt = 46.00 Mean = 36 Exp = 1.841 X/Y/Z = -0.369 0.360 0.842 106850.906: Saved state 54 at 46.0: def 0.742 IS 0.144 0.517 int 0.12073 exp 1.841 106850.922: SetIntensity called with factor 1.0135 106850.937: Changing (ideal) Record exposure by 1.0135 to 1.8657 106850.953: Frame time for constraint is 0.1598 106850.969: Will be constrained to 1.9174 (frame 0.1598, constraint 0.1598) 106854.031: recentlyReversed: dir 1 ang 47.999119 revtilt 50.998252 back 3.000000 3mte 0.300000 Predicted X = -0.353 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Y = 0.342 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Z = 0.697 StdErr = 0.000 nFit = 2 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.010, 0.003 um; position -0.363, 0.340 Measured defocus = 1.10 microns changed by -1.20 to target Measured defocus = -1.04 microns changed by 0.94 to target Tracking image shifted 0.000, -0.003 um; position -0.363, 0.342 12 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0055_48.0.tif Frame alignment results: distance raw = 30.5 smoothed = 29.7 Weighted resid mean = 0.31 max max = 0.54 Mean unweighted max = 30.09 Record image shifted 0.000, -0.006 um Tracking reference shifted by 0.6,4.1 pixels, 0.000,-0.006 um Saved Z = 55 at 28.82 min Tilt = 48.00 Mean = 35 Exp = 1.866 X/Y/Z = -0.364 0.348 0.435 106886.125: Saved state 55 at 48.0: def 0.335 IS 0.145 0.503 int 0.12073 exp 1.866 106886.140: SetIntensity called with factor 1.0146 106886.156: Changing (ideal) Record exposure by 1.0146 to 1.8930 106886.172: Frame time for constraint is 0.1598 106886.187: Will be constrained to 1.9174 (frame 0.1598, constraint 0.1598) 106889.906: recentlyReversed: dir 1 ang 49.999874 revtilt 53.001007 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.011 0.006 um, 0.8 0.4 % Error in Z Prediction -0.261 Predicted X = -0.351 StdErr = 0.008 nFit = 3 nDrop = 0 Predicted Y = 0.331 StdErr = 0.004 nFit = 3 nDrop = 0 Predicted Z = 0.203 StdErr = 0.195 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.009, 0.033 um; position -0.359, 0.299 Measured defocus = 0.05 microns changed by -0.15 to target Tracking image shifted 0.000, -0.002 um; position -0.360, 0.300 12 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070_0056_50.0.tif Frame alignment results: distance raw = 28.7 smoothed = 28.1 Weighted resid mean = 0.30 max max = 0.82 Mean unweighted max = 28.21 Record image shifted -0.000, -0.008 um Tracking reference shifted by 1.1,5.8 pixels, -0.000,-0.008 um Saved Z = 56 at 29.35 min Tilt = 50.00 Mean = 33 Exp = 1.893 X/Y/Z = -0.359 0.308 0.054 2021/11/19 20:38 W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_070.mrc 0 57 11520 8184 10747 -10.0 50.0 2.00 REG 29.4 1x1 Continuum_K3 26000 1 0.17 LD-DS 20.0 0.0 Tilt series acquired at item # 29 with label 49 (item at 215.34, -317.60, TS at 215.42, -316.00) ################# PostTomo Started a long running operation: updating hardware dark reference Call for updating hardware dark reference finished successfully ################# PostTomo END Focus area changed from stored position for item, axis position -4.00 angle 0, subarea offset 0 0