18-Nov-21 17:07:16 Rough eucentricity: changing Z by -1.89 to -60.08, finished. 7836.562: (Initial) alignment to map 29 (43) 7836.562: Target X, Y: -357.141 185.333 7836.562: Map matches low dose View; staying in low dose but with exp 0.405 intensity 0.19459 bin 4 Trimming A by 85 122 105 130, reference by 80 127 105 120 Disparity in stage position after aligning to center of map frame: 0.03 0.38 Aligning to target with image shift equivalent to stage shift: 0.03 0.38 7843.672: Map matches low dose Preview; staying in low dose but with exp 0.202 intensity 0.12073 bin 4 7855.687: Realigned to map item itself with image shift of 31.8 141.9 pixels Disparity in stage position after aligning to map item itself: -0.02 0.60 (align shift -0.06 0.19) Finished aligning to item in 2 rounds Opened new file W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037.mrc STARTING TILT SERIES WITH AUTOMATIC TERMINATION AFTER ERRORS. 7863.469: Skipping reverse tilt: dir -1 start 0.00 revang 3.00 revtilt 2.98 maxtilt 69.90 maxerr 0.10 Measured defocus = -8.43 microns changed by 8.33 to target Measured defocus = -0.17 microns changed by 0.07 to target 7898.125: SetIntensity called with factor 0.9962 7898.125: Changing (ideal) Record exposure by 0.9962 to 1.5918 7898.125: Frame time for constraint is 0.1598 7898.125: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 7898.125: Skipping reverse tilt: dir -1 start -10.00 revang -7.00 revtilt 2.98 maxtilt 69.90 maxerr 0.10 Need regular tracking - Need autofocus Tracking image shifted 0.001, -0.025 um; position 0.071, -0.156 Measured defocus = -0.05 microns changed by -0.05 to target Tracking image shifted 0.001, -0.002 um; position 0.070, -0.154 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0000_-10.0.tif Frame alignment results: distance raw = 14.1 smoothed = 12.8 Weighted resid mean = 0.06 max max = 0.12 Mean unweighted max = 0.55 Saved Z = 0 at 0.00 min Tilt = -10.00 Mean = 47 Exp = 1.592 X/Y/Z = 0.070 -0.154 3.453 7917.922: Saved state 0 at -10.0: def 3.353 IS 0.013 -0.174 int 0.12073 exp 1.592 7917.937: SetIntensity called with factor 1.0002 7917.937: Changing (ideal) Record exposure by 1.0002 to 1.5920 7917.937: Frame time for constraint is 0.1598 7917.937: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Need regular tracking - Need autofocus Tracking image shifted -0.008, 0.004 um; position 0.079, -0.158 Measured defocus = -0.23 microns changed by 0.13 to target Tracking image shifted 0.005, -0.006 um; position 0.074, -0.152 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0001_-12.0.tif Frame alignment results: distance raw = 11.4 smoothed = 10.4 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.37 Record image shifted 0.002, -0.011 um Tracking reference shifted by 0.5,8.5 pixels, 0.002,-0.011 um Saved Z = 1 at 0.47 min Tilt = -11.99 Mean = 47 Exp = 1.592 X/Y/Z = 0.072 -0.141 3.584 7946.390: Saved state 1 at -12.0: def 3.484 IS 0.005 -0.163 int 0.12073 exp 1.592 7946.406: SetIntensity called with factor 1.0005 7946.406: Changing (ideal) Record exposure by 1.0005 to 1.5927 7946.406: Frame time for constraint is 0.1598 7946.406: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Predicted X = 0.074 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Y = -0.127 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Z = 3.716 StdErr = 0.000 nFit = 2 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.003, -0.021 um; position 0.072, -0.106 Measured defocus = -0.02 microns changed by -0.08 to target Tracking image shifted 0.004, -0.004 um; position 0.068, -0.102 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0002_-14.0.tif Frame alignment results: distance raw = 11.0 smoothed = 10.1 Weighted resid mean = 0.07 max max = 0.10 Mean unweighted max = 0.19 Record image shifted 0.001, -0.010 um Tracking reference shifted by 0.3,7.5 pixels, 0.001,-0.010 um Saved Z = 2 at 0.96 min Tilt = -13.99 Mean = 47 Exp = 1.593 X/Y/Z = 0.066 -0.092 3.639 7975.453: Saved state 2 at -14.0: def 3.539 IS -0.013 -0.117 int 0.12073 exp 1.593 7975.531: Restored state 0 at -10.0 for tiltInd 3 def 3.353 IS 0.013 -0.174 7975.547: SetIntensity called with factor 1.0002 7975.547: Changing (ideal) Record exposure by 1.0002 to 1.5920 7975.547: Frame time for constraint is 0.1598 7975.547: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 7979.765: recentlyReversed: dir 1 ang -7.998020 revtilt -5.000387 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.008 0.035 um, 0.6 2.5 % Error in Z Prediction -0.077 Need regular tracking - Need autofocus Tracking image shifted 0.063, -0.106 um; position 0.007, -0.048 Measured defocus = -0.00 microns changed by -0.10 to target Tracking image shifted 0.004, -0.006 um; position 0.003, -0.042 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0003_-8.0.tif Frame alignment results: distance raw = 12.2 smoothed = 11.2 Weighted resid mean = 0.07 max max = 0.12 Mean unweighted max = 0.39 Record image shifted 0.000, 0.006 um Tracking reference shifted by -1.1,-4.0 pixels, 0.000,0.006 um Saved Z = 3 at 1.49 min Tilt = -8.00 Mean = 47 Exp = 1.592 X/Y/Z = 0.003 -0.048 3.358 8007.265: Saved state 3 at -8.0: def 3.258 IS 0.020 -0.043 int 0.12073 exp 1.592 8007.265: SetIntensity called with factor 1.0005 8007.281: Changing (ideal) Record exposure by 1.0005 to 1.5927 8007.281: Frame time for constraint is 0.1598 8007.281: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8010.687: recentlyReversed: dir 1 ang -5.999265 revtilt -3.000632 back 3.000000 3mte 0.300000 Need regular tracking - Need autofocus Tracking image shifted 0.003, 0.003 um; position -0.000, -0.051 Measured defocus = -0.02 microns changed by -0.08 to target Tracking image shifted 0.003, -0.006 um; position -0.003, -0.045 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0004_-6.0.tif Frame alignment results: distance raw = 12.3 smoothed = 11.3 Weighted resid mean = 0.07 max max = 0.12 Mean unweighted max = 0.44 Record image shifted 0.000, 0.008 um Tracking reference shifted by -1.2,-6.1 pixels, 0.000,0.008 um Saved Z = 4 at 2.01 min Tilt = -6.00 Mean = 47 Exp = 1.593 X/Y/Z = -0.003 -0.054 3.275 8038.656: Saved state 4 at -6.0: def 3.175 IS 0.028 -0.046 int 0.12073 exp 1.593 8038.734: Restored state 2 at -14.0 for tiltInd 5 def 3.539 IS -0.013 -0.117 8038.734: SetIntensity called with factor 1.0008 8038.734: Changing (ideal) Record exposure by 1.0008 to 1.5940 8038.734: Frame time for constraint is 0.1598 8038.750: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Predicted X = 0.065 StdErr = 0.006 nFit = 3 nDrop = 0 Predicted Y = -0.067 StdErr = 0.026 nFit = 3 nDrop = 0 Predicted Z = 3.745 StdErr = 0.057 nFit = 3 nDrop = 0 Need autofocus Measured defocus = -0.08 microns changed by -0.02 to target 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0005_-16.0.tif Frame alignment results: distance raw = 9.9 smoothed = 9.2 Weighted resid mean = 0.06 max max = 0.11 Mean unweighted max = 0.29 Record image shifted 0.062, -0.067 um Saved Z = 5 at 2.48 min Tilt = -16.00 Mean = 47 Exp = 1.594 X/Y/Z = 0.004 -0.000 3.726 8067.031: Saved state 5 at -16.0: def 3.626 IS -0.003 -0.002 int 0.12073 exp 1.594 8067.031: SetIntensity called with factor 1.0011 8067.047: Changing (ideal) Record exposure by 1.0011 to 1.5957 8067.047: Frame time for constraint is 0.1598 8067.047: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.062 0.067 um, 4.5 4.8 % Error in Z Prediction -0.019 Predicted X = -0.021 StdErr = 0.042 nFit = 3 nDrop = 0 Predicted Y = 0.063 StdErr = 0.032 nFit = 3 nDrop = 0 Predicted Z = 3.791 StdErr = 0.024 nFit = 3 nDrop = 0 Need regular tracking - Tracking image shifted -0.023, -0.016 um; position 0.001, 0.079 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0006_-18.0.tif Frame alignment results: distance raw = 8.1 smoothed = 7.3 Weighted resid mean = 0.08 max max = 0.13 Mean unweighted max = 0.18 Record image shifted 0.005, -0.018 um Tracking reference shifted by -1.0,13.7 pixels, 0.005,-0.018 um Saved Z = 6 at 2.87 min Tilt = -17.99 Mean = 47 Exp = 1.596 X/Y/Z = -0.004 0.097 3.791 8090.281: Saved state 6 at -18.0: def 3.691 IS -0.043 0.087 int 0.12073 exp 1.596 8090.359: Restored state 4 at -6.0 for tiltInd 7 def 3.175 IS 0.028 -0.046 8090.359: SetIntensity called with factor 1.0008 8090.375: Changing (ideal) Record exposure by 1.0008 to 1.5940 8090.375: Frame time for constraint is 0.1598 8090.375: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8095.125: recentlyReversed: dir 1 ang -3.998510 revtilt -1.005377 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.018 0.034 um, 1.3 2.5 % Predicted X = -0.009 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Y = -0.059 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Z = 3.192 StdErr = 0.000 nFit = 2 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.007, 0.019 um; position -0.016, -0.078 Measured defocus = -0.03 microns changed by -0.07 to target Tracking image shifted 0.002, -0.006 um; position -0.018, -0.072 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0007_-4.0.tif Frame alignment results: distance raw = 11.5 smoothed = 10.6 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.23 Record image shifted -0.001, 0.008 um Tracking reference shifted by -0.3,-5.9 pixels, -0.001,0.008 um Saved Z = 7 at 3.42 min Tilt = -4.00 Mean = 47 Exp = 1.594 X/Y/Z = -0.017 -0.080 3.121 8122.969: Saved state 7 at -4.0: def 3.021 IS 0.053 -0.062 int 0.12073 exp 1.594 8122.984: SetIntensity called with factor 1.0011 8122.984: Changing (ideal) Record exposure by 1.0011 to 1.5957 8122.984: Frame time for constraint is 0.1598 8123.000: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8126.359: recentlyReversed: dir 1 ang -1.998755 revtilt 0.994378 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.008 -0.021 um, 0.6 1.5 % Error in Z Prediction -0.071 Predicted X = -0.026 StdErr = 0.006 nFit = 3 nDrop = 0 Predicted Y = -0.093 StdErr = 0.015 nFit = 3 nDrop = 0 Predicted Z = 3.015 StdErr = 0.053 nFit = 3 nDrop = 0 Need regular tracking - Tracking image shifted -0.014, 0.060 um; position -0.011, -0.153 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0008_-2.0.tif Frame alignment results: distance raw = 9.7 smoothed = 8.5 Weighted resid mean = 0.06 max max = 0.09 Mean unweighted max = 0.21 Record image shifted 0.000, 0.005 um Tracking reference shifted by -1.1,-3.9 pixels, 0.000,0.005 um Saved Z = 8 at 3.84 min Tilt = -2.00 Mean = 47 Exp = 1.596 X/Y/Z = -0.012 -0.158 3.015 8148.656: Saved state 8 at -2.0: def 2.915 IS 0.085 -0.134 int 0.12073 exp 1.596 8148.734: Restored state 6 at -18.0 for tiltInd 9 def 3.691 IS -0.043 0.087 8148.734: SetIntensity called with factor 1.0014 8148.734: Changing (ideal) Record exposure by 1.0014 to 1.5979 8148.734: Frame time for constraint is 0.1598 8148.750: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.014 -0.065 um, 1.0 4.7 % Predicted X = -0.048 StdErr = 0.041 nFit = 3 nDrop = 0 Predicted Y = 0.190 StdErr = 0.004 nFit = 3 nDrop = 0 Predicted Z = 3.862 StdErr = 0.031 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.024, 0.038 um; position -0.023, 0.152 Measured defocus = -0.05 microns changed by -0.05 to target Tracking image shifted 0.004, 0.000 um; position -0.028, 0.151 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0009_-20.0.tif Frame alignment results: distance raw = 7.9 smoothed = 7.2 Weighted resid mean = 0.08 max max = 0.11 Mean unweighted max = 0.33 Record image shifted -0.002, -0.005 um Tracking reference shifted by 2.3,3.2 pixels, -0.002,-0.005 um Saved Z = 9 at 4.36 min Tilt = -19.99 Mean = 47 Exp = 1.598 X/Y/Z = -0.025 0.156 3.808 8179.875: Saved state 9 at -20.0: def 3.708 IS -0.053 0.152 int 0.12073 exp 1.598 8179.890: SetIntensity called with factor 1.0017 8179.890: Changing (ideal) Record exposure by 1.0017 to 1.6006 8179.890: Frame time for constraint is 0.1598 8179.906: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.022 -0.034 um, 1.6 2.5 % Error in Z Prediction -0.054 Predicted X = -0.038 StdErr = 0.011 nFit = 3 nDrop = 0 Predicted Y = 0.241 StdErr = 0.028 nFit = 3 nDrop = 0 Predicted Z = 3.868 StdErr = 0.041 nFit = 3 nDrop = 0 Need regular tracking - Tracking image shifted -0.020, 0.087 um; position -0.018, 0.154 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0010_-22.0.tif Frame alignment results: distance raw = 7.7 smoothed = 7.1 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.21 Record image shifted 0.003, -0.004 um Tracking reference shifted by -1.7,3.6 pixels, 0.003,-0.004 um Saved Z = 10 at 4.76 min Tilt = -21.99 Mean = 47 Exp = 1.601 X/Y/Z = -0.021 0.158 3.868 8203.359: Saved state 10 at -22.0: def 3.768 IS -0.057 0.151 int 0.12073 exp 1.601 8203.437: Restored state 8 at -2.0 for tiltInd 11 def 2.915 IS 0.085 -0.134 8203.437: SetIntensity called with factor 1.0014 8203.437: Changing (ideal) Record exposure by 1.0014 to 1.5979 8203.453: Frame time for constraint is 0.1598 8203.453: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8208.844: recentlyReversed: dir 1 ang 0.001000 revtilt 2.993633 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.016 -0.083 um, 1.2 6.0 % Predicted X = -0.019 StdErr = 0.014 nFit = 3 nDrop = 0 Predicted Y = -0.202 StdErr = 0.038 nFit = 3 nDrop = 0 Predicted Z = 2.897 StdErr = 0.070 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.007, -0.007 um; position -0.026, -0.195 Measured defocus = -0.29 microns changed by 0.19 to target Tracking image shifted 0.002, -0.009 um; position -0.029, -0.186 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0011_0.0.tif Frame alignment results: distance raw = 13.2 smoothed = 12.1 Weighted resid mean = 0.06 max max = 0.11 Mean unweighted max = 0.37 Record image shifted -0.004, 0.016 um Tracking reference shifted by 0.9,-12.1 pixels, -0.004,0.016 um Saved Z = 11 at 5.31 min Tilt = 0.00 Mean = 47 Exp = 1.598 X/Y/Z = -0.025 -0.202 3.090 8236.594: Saved state 11 at 0.0: def 2.990 IS 0.117 -0.165 int 0.12073 exp 1.598 8236.609: SetIntensity called with factor 1.0017 8236.609: Changing (ideal) Record exposure by 1.0017 to 1.6006 8236.609: Frame time for constraint is 0.1598 8236.625: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8240.015: recentlyReversed: dir 1 ang 1.999255 revtilt 4.993888 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.005 0.000 um, 0.4 0.0 % Error in Z Prediction 0.194 Predicted X = -0.025 StdErr = 0.013 nFit = 3 nDrop = 0 Predicted Y = -0.269 StdErr = 0.026 nFit = 3 nDrop = 0 Predicted Z = 3.015 StdErr = 0.125 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.003, -0.054 um; position -0.023, -0.215 Measured defocus = 0.01 microns changed by -0.11 to target Tracking image shifted 0.003, -0.008 um; position -0.026, -0.207 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0012_2.0.tif Frame alignment results: distance raw = 13.4 smoothed = 12.3 Weighted resid mean = 0.06 max max = 0.09 Mean unweighted max = 0.28 Record image shifted -0.002, 0.010 um Tracking reference shifted by -0.2,-7.2 pixels, -0.002,0.010 um Saved Z = 12 at 5.83 min Tilt = 2.00 Mean = 46 Exp = 1.601 X/Y/Z = -0.024 -0.217 2.903 8267.531: Saved state 12 at 2.0: def 2.803 IS 0.123 -0.178 int 0.12073 exp 1.601 8267.609: Restored state 10 at -22.0 for tiltInd 13 def 3.768 IS -0.057 0.151 8267.609: SetIntensity called with factor 1.0020 8267.625: Changing (ideal) Record exposure by 1.0020 to 1.6039 8267.625: Frame time for constraint is 0.1598 8267.640: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.001 0.052 um, 0.1 3.8 % Error in Z Prediction -0.113 Predicted X = -0.021 StdErr = 0.027 nFit = 4 nDrop = 0 Predicted Y = 0.117 StdErr = 0.014 nFit = 4 nDrop = 0 Predicted Z = 3.922 StdErr = 0.050 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.013, -0.047 um; position -0.034, 0.164 Measured defocus = -0.10 microns changed by -0.00 to target Tracking image shifted 0.003, 0.005 um; position -0.038, 0.159 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0013_-24.0.tif Frame alignment results: distance raw = 9.5 smoothed = 8.9 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.27 Record image shifted -0.002, -0.009 um Tracking reference shifted by 2.7,6.5 pixels, -0.002,-0.009 um Saved Z = 13 at 6.35 min Tilt = -23.99 Mean = 47 Exp = 1.604 X/Y/Z = -0.036 0.168 3.916 8298.953: Saved state 13 at -24.0: def 3.816 IS -0.049 0.168 int 0.12073 exp 1.604 8298.953: SetIntensity called with factor 1.0024 8298.969: Changing (ideal) Record exposure by 1.0024 to 1.6076 8298.969: Frame time for constraint is 0.1598 8298.984: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.015 0.051 um, 1.1 3.7 % Error in Z Prediction -0.006 Predicted X = -0.038 StdErr = 0.014 nFit = 3 nDrop = 0 Predicted Y = 0.120 StdErr = 0.030 nFit = 5 nDrop = 0 Predicted Z = 3.970 StdErr = 0.026 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted -0.014, -0.014 um; position -0.024, 0.134 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0014_-26.0.tif Frame alignment results: distance raw = 9.8 smoothed = 8.7 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.38 Record image shifted 0.003, -0.001 um Tracking reference shifted by -2.4,1.4 pixels, 0.003,-0.001 um Saved Z = 14 at 6.74 min Tilt = -25.99 Mean = 47 Exp = 1.608 X/Y/Z = -0.027 0.135 3.970 8322.515: Saved state 14 at -26.0: def 3.870 IS -0.041 0.134 int 0.12073 exp 1.608 8322.594: Restored state 12 at 2.0 for tiltInd 15 def 2.803 IS 0.123 -0.178 8322.594: SetIntensity called with factor 1.0020 8322.594: Changing (ideal) Record exposure by 1.0020 to 1.6038 8322.609: Frame time for constraint is 0.1598 8322.609: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8328.531: recentlyReversed: dir 1 ang 3.999010 revtilt 6.998143 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.010 0.016 um, 0.7 1.1 % Predicted X = -0.032 StdErr = 0.010 nFit = 3 nDrop = 0 Predicted Y = -0.251 StdErr = 0.022 nFit = 3 nDrop = 0 Predicted Z = 2.887 StdErr = 0.145 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.008, -0.040 um; position -0.041, -0.211 Measured defocus = 0.05 microns changed by -0.15 to target Tracking image shifted 0.001, -0.005 um; position -0.042, -0.206 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0015_4.0.tif Frame alignment results: distance raw = 14.7 smoothed = 13.7 Weighted resid mean = 0.08 max max = 0.13 Mean unweighted max = 0.38 Record image shifted -0.001, 0.008 um Tracking reference shifted by -0.6,-6.0 pixels, -0.001,0.008 um Saved Z = 15 at 7.30 min Tilt = 4.00 Mean = 46 Exp = 1.604 X/Y/Z = -0.041 -0.215 2.733 8356.094: Saved state 15 at 4.0: def 2.633 IS 0.137 -0.168 int 0.12073 exp 1.604 8356.094: SetIntensity called with factor 1.0023 8356.109: Changing (ideal) Record exposure by 1.0023 to 1.6076 8356.109: Frame time for constraint is 0.1598 8356.125: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8359.953: recentlyReversed: dir 1 ang 5.998765 revtilt 8.999397 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.009 0.036 um, 0.6 2.6 % Error in Z Prediction -0.154 Predicted X = -0.052 StdErr = 0.020 nFit = 4 nDrop = 0 Predicted Y = -0.187 StdErr = 0.008 nFit = 4 nDrop = 0 Predicted Z = 2.699 StdErr = 0.137 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.015, 0.042 um; position -0.038, -0.229 Measured defocus = -0.11 microns changed by 0.01 to target Tracking image shifted 0.002, -0.005 um; position -0.039, -0.223 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0016_6.0.tif Frame alignment results: distance raw = 13.0 smoothed = 11.9 Weighted resid mean = 0.09 max max = 0.15 Mean unweighted max = 0.31 Record image shifted -0.001, 0.012 um Tracking reference shifted by -0.5,-8.8 pixels, -0.001,0.012 um Saved Z = 16 at 7.83 min Tilt = 6.00 Mean = 46 Exp = 1.608 X/Y/Z = -0.038 -0.235 2.711 8387.734: Saved state 16 at 6.0: def 2.611 IS 0.144 -0.188 int 0.12073 exp 1.608 8387.812: Restored state 14 at -26.0 for tiltInd 17 def 3.870 IS -0.041 0.134 8387.828: SetIntensity called with factor 1.0027 8387.828: Changing (ideal) Record exposure by 1.0027 to 1.6120 8387.828: Frame time for constraint is 0.1598 8387.844: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.015 -0.049 um, 1.1 3.5 % Error in Z Prediction 0.012 Predicted X = -0.028 StdErr = 0.028 nFit = 4 nDrop = 0 Predicted Y = 0.067 StdErr = 0.022 nFit = 6 nDrop = 0 Predicted Z = 4.023 StdErr = 0.029 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.014, -0.090 um; position -0.042, 0.157 Measured defocus = -0.21 microns changed by 0.11 to target Tracking image shifted 0.003, 0.006 um; position -0.045, 0.151 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0017_-28.0.tif Frame alignment results: distance raw = 8.9 smoothed = 8.3 Weighted resid mean = 0.06 max max = 0.11 Mean unweighted max = 0.33 Record image shifted -0.002, -0.008 um Tracking reference shifted by 2.4,5.6 pixels, -0.002,-0.008 um Saved Z = 17 at 8.37 min Tilt = -27.99 Mean = 47 Exp = 1.612 X/Y/Z = -0.043 0.159 4.126 8420.328: Saved state 17 at -28.0: def 4.026 IS -0.039 0.163 int 0.12073 exp 1.612 8420.344: SetIntensity called with factor 1.0030 8420.344: Changing (ideal) Record exposure by 1.0030 to 1.6168 8420.359: Frame time for constraint is 0.1598 8420.359: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.015 0.092 um, 1.1 6.6 % Error in Z Prediction 0.103 Predicted X = -0.048 StdErr = 0.031 nFit = 4 nDrop = 0 Predicted Y = 0.150 StdErr = 0.040 nFit = 5 nDrop = 0 Predicted Z = 4.156 StdErr = 0.067 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.005, -0.085 um; position -0.042, 0.235 Measured defocus = -0.02 microns changed by -0.08 to target Tracking image shifted 0.003, 0.007 um; position -0.045, 0.228 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0018_-30.0.tif Frame alignment results: distance raw = 12.8 smoothed = 12.1 Weighted resid mean = 0.06 max max = 0.11 Mean unweighted max = 0.31 Record image shifted 0.001, -0.006 um Tracking reference shifted by 0.2,4.4 pixels, 0.001,-0.006 um Saved Z = 18 at 8.86 min Tilt = -29.99 Mean = 47 Exp = 1.617 X/Y/Z = -0.045 0.234 4.073 8449.453: Saved state 18 at -30.0: def 3.973 IS -0.072 0.231 int 0.12073 exp 1.617 8449.515: Restored state 16 at 6.0 for tiltInd 19 def 2.611 IS 0.144 -0.188 8449.515: SetIntensity called with factor 1.0027 8449.531: Changing (ideal) Record exposure by 1.0027 to 1.6119 8449.531: Frame time for constraint is 0.1598 8449.547: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8456.422: recentlyReversed: dir 1 ang 7.999520 revtilt 10.998653 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.002 0.084 um, 0.2 6.0 % Error in Z Prediction -0.083 Predicted X = -0.042 StdErr = 0.025 nFit = 4 nDrop = 0 Predicted Y = -0.224 StdErr = 0.028 nFit = 5 nDrop = 0 Predicted Z = 2.612 StdErr = 0.106 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.008, 0.034 um; position -0.050, -0.258 Measured defocus = -0.03 microns changed by -0.07 to target Tracking image shifted 0.001, -0.001 um; position -0.051, -0.257 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0019_8.0.tif Frame alignment results: distance raw = 18.2 smoothed = 17.1 Weighted resid mean = 0.06 max max = 0.11 Mean unweighted max = 0.36 Record image shifted -0.002, 0.010 um Tracking reference shifted by 0.1,-7.6 pixels, -0.002,0.010 um Saved Z = 19 at 9.43 min Tilt = 8.00 Mean = 46 Exp = 1.612 X/Y/Z = -0.049 -0.267 2.542 8483.969: Saved state 19 at 8.0: def 2.442 IS 0.168 -0.209 int 0.12073 exp 1.612 8483.984: SetIntensity called with factor 1.0030 8483.984: Changing (ideal) Record exposure by 1.0030 to 1.6167 8484.000: Frame time for constraint is 0.1598 8484.000: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8487.390: recentlyReversed: dir 1 ang 9.999275 revtilt 12.978910 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.007 -0.043 um, 0.5 3.1 % Error in Z Prediction -0.070 Predicted X = -0.051 StdErr = 0.011 nFit = 3 nDrop = 0 Predicted Y = -0.299 StdErr = 0.018 nFit = 5 nDrop = 1 Predicted Z = 2.477 StdErr = 0.095 nFit = 6 nDrop = 0 Need regular tracking - Tracking image shifted -0.012, 0.031 um; position -0.039, -0.331 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0020_10.0.tif Frame alignment results: distance raw = 12.3 smoothed = 10.9 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.17 Record image shifted 0.001, 0.009 um Tracking reference shifted by -1.7,-6.6 pixels, 0.001,0.009 um Saved Z = 20 at 9.86 min Tilt = 10.00 Mean = 46 Exp = 1.617 X/Y/Z = -0.039 -0.340 2.477 8509.656: Saved state 20 at 10.0: def 2.377 IS 0.195 -0.279 int 0.12073 exp 1.617 8509.734: Restored state 18 at -30.0 for tiltInd 21 def 3.973 IS -0.072 0.231 8509.750: SetIntensity called with factor 1.0034 8509.750: Changing (ideal) Record exposure by 1.0034 to 1.6223 8509.765: Frame time for constraint is 0.1598 8509.765: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.011 -0.041 um, 0.8 2.9 % Predicted X = -0.057 StdErr = 0.010 nFit = 3 nDrop = 0 Predicted Y = 0.274 StdErr = 0.050 nFit = 6 nDrop = 0 Predicted Z = 4.182 StdErr = 0.084 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.004, -0.006 um; position -0.060, 0.280 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0021_-32.0.tif Frame alignment results: distance raw = 8.6 smoothed = 7.8 Weighted resid mean = 0.05 max max = 0.10 Mean unweighted max = 0.31 Record image shifted 0.002, 0.002 um Tracking reference shifted by -1.9,-1.3 pixels, 0.002,0.002 um Saved Z = 21 at 10.32 min Tilt = -31.99 Mean = 47 Exp = 1.622 X/Y/Z = -0.063 0.277 4.182 8537.297: Saved state 21 at -32.0: def 4.082 IS -0.078 0.279 int 0.12073 exp 1.622 8537.297: SetIntensity called with factor 1.0037 8537.312: Changing (ideal) Record exposure by 1.0037 to 1.6283 8537.312: Frame time for constraint is 0.1598 8537.328: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.006 0.003 um, 0.4 0.3 % Predicted X = -0.070 StdErr = 0.011 nFit = 3 nDrop = 0 Predicted Y = 0.392 StdErr = 0.049 nFit = 5 nDrop = 0 Predicted Z = 4.244 StdErr = 0.095 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.010, 0.106 um; position -0.059, 0.286 Measured defocus = -0.13 microns changed by 0.03 to target Tracking image shifted 0.002, 0.005 um; position -0.061, 0.281 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0022_-34.0.tif Frame alignment results: distance raw = 11.1 smoothed = 10.3 Weighted resid mean = 0.08 max max = 0.14 Mean unweighted max = 0.21 Record image shifted -0.001, -0.006 um Tracking reference shifted by 1.6,4.5 pixels, -0.001,-0.006 um Saved Z = 22 at 10.81 min Tilt = -33.99 Mean = 47 Exp = 1.628 X/Y/Z = -0.060 0.287 4.266 8566.625: Saved state 22 at -34.0: def 4.166 IS -0.085 0.286 int 0.12073 exp 1.628 8566.719: Restored state 20 at 10.0 for tiltInd 23 def 2.377 IS 0.195 -0.279 8566.734: SetIntensity called with factor 1.0034 8566.734: Changing (ideal) Record exposure by 1.0034 to 1.6221 8566.750: Frame time for constraint is 0.1598 8566.750: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8573.937: recentlyReversed: dir 1 ang 11.999530 revtilt 14.995163 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.010 -0.104 um, 0.7 7.5 % Error in Z Prediction 0.022 Predicted X = -0.044 StdErr = 0.016 nFit = 3 nDrop = 0 Predicted Y = -0.425 StdErr = 0.013 nFit = 5 nDrop = 1 Predicted Z = 2.376 StdErr = 0.104 nFit = 6 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.006, -0.039 um; position -0.050, -0.386 Measured defocus = -0.09 microns changed by -0.01 to target Tracking image shifted -0.001, 0.001 um; position -0.048, -0.387 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0023_12.0.tif Frame alignment results: distance raw = 16.7 smoothed = 15.8 Weighted resid mean = 0.09 max max = 0.15 Mean unweighted max = 0.34 Record image shifted -0.003, 0.012 um Tracking reference shifted by 0.4,-9.0 pixels, -0.003,0.012 um Saved Z = 23 at 11.40 min Tilt = 12.00 Mean = 45 Exp = 1.622 X/Y/Z = -0.046 -0.399 2.370 8601.734: Saved state 23 at 12.0: def 2.270 IS 0.228 -0.328 int 0.12073 exp 1.622 8601.734: SetIntensity called with factor 1.0037 8601.750: Changing (ideal) Record exposure by 1.0037 to 1.6282 8601.765: Frame time for constraint is 0.1598 8601.765: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8605.062: recentlyReversed: dir 1 ang 13.999285 revtilt 16.998417 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.002 0.026 um, 0.2 1.9 % Error in Z Prediction -0.006 Predicted X = -0.041 StdErr = 0.012 nFit = 3 nDrop = 0 Predicted Y = -0.492 StdErr = 0.022 nFit = 5 nDrop = 0 Predicted Z = 2.222 StdErr = 0.072 nFit = 6 nDrop = 0 Need regular tracking - Tracking image shifted -0.006, -0.047 um; position -0.036, -0.445 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0024_14.0.tif Frame alignment results: distance raw = 10.5 smoothed = 9.6 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.35 Record image shifted -0.001, 0.011 um Tracking reference shifted by -0.9,-7.9 pixels, -0.001,0.011 um Saved Z = 24 at 11.82 min Tilt = 14.00 Mean = 45 Exp = 1.628 X/Y/Z = -0.035 -0.456 2.222 8627.390: Saved state 24 at 14.0: def 2.122 IS 0.246 -0.384 int 0.12073 exp 1.628 8627.484: Restored state 22 at -34.0 for tiltInd 25 def 4.166 IS -0.085 0.286 8627.484: SetIntensity called with factor 1.0041 8627.500: Changing (ideal) Record exposure by 1.0041 to 1.6350 8627.500: Frame time for constraint is 0.1598 8627.515: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.006 0.037 um, 0.5 2.6 % Predicted X = -0.063 StdErr = 0.023 nFit = 4 nDrop = 0 Predicted Y = 0.316 StdErr = 0.048 nFit = 5 nDrop = 1 Predicted Z = 4.324 StdErr = 0.090 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.010, 0.025 um; position -0.073, 0.292 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0025_-36.0.tif Frame alignment results: distance raw = 6.8 smoothed = 6.0 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.31 Record image shifted 0.002, 0.004 um Tracking reference shifted by -1.8,-2.9 pixels, 0.002,0.004 um Saved Z = 25 at 12.29 min Tilt = -35.99 Mean = 47 Exp = 1.635 X/Y/Z = -0.074 0.287 4.324 8655.234: Saved state 25 at -36.0: def 4.224 IS -0.073 0.294 int 0.12073 exp 1.635 8655.250: SetIntensity called with factor 1.0045 8655.250: Changing (ideal) Record exposure by 1.0045 to 1.6424 8655.265: Frame time for constraint is 0.1598 8655.265: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.011 -0.029 um, 0.8 2.1 % Predicted X = -0.078 StdErr = 0.025 nFit = 4 nDrop = 0 Predicted Y = 0.251 StdErr = 0.012 nFit = 5 nDrop = 0 Predicted Z = 4.389 StdErr = 0.102 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.011, -0.026 um; position -0.067, 0.277 Measured defocus = -0.37 microns changed by 0.27 to target Tracking image shifted -0.000, -0.000 um; position -0.067, 0.277 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0026_-38.0.tif Frame alignment results: distance raw = 7.4 smoothed = 6.9 Weighted resid mean = 0.06 max max = 0.11 Mean unweighted max = 0.29 Record image shifted 0.001, -0.000 um Tracking reference shifted by -0.4,0.3 pixels, 0.001,-0.000 um Saved Z = 26 at 12.77 min Tilt = -37.99 Mean = 47 Exp = 1.642 X/Y/Z = -0.068 0.277 4.656 8684.234: Saved state 26 at -38.0: def 4.556 IS -0.074 0.281 int 0.12073 exp 1.642 8684.344: Restored state 24 at 14.0 for tiltInd 27 def 2.122 IS 0.246 -0.384 8684.344: SetIntensity called with factor 1.0041 8684.359: Changing (ideal) Record exposure by 1.0041 to 1.6348 8684.359: Frame time for constraint is 0.1598 8684.375: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8691.828: recentlyReversed: dir 1 ang 15.999040 revtilt 18.998672 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.011 0.027 um, 0.8 1.9 % Error in Z Prediction 0.267 Predicted X = -0.032 StdErr = 0.022 nFit = 4 nDrop = 0 Predicted Y = -0.529 StdErr = 0.027 nFit = 5 nDrop = 0 Predicted Z = 2.106 StdErr = 0.079 nFit = 6 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.020, -0.088 um; position -0.052, -0.441 Measured defocus = 0.11 microns changed by -0.21 to target Tracking image shifted -0.001, 0.001 um; position -0.051, -0.442 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0027_16.0.tif Frame alignment results: distance raw = 12.0 smoothed = 11.2 Weighted resid mean = 0.06 max max = 0.11 Mean unweighted max = 0.41 Record image shifted -0.003, 0.010 um Tracking reference shifted by 1.1,-7.8 pixels, -0.003,0.010 um Saved Z = 27 at 13.36 min Tilt = 16.00 Mean = 45 Exp = 1.635 X/Y/Z = -0.048 -0.452 1.893 8719.531: Saved state 27 at 16.0: def 1.793 IS 0.256 -0.374 int 0.12073 exp 1.635 8719.531: SetIntensity called with factor 1.0045 8719.547: Changing (ideal) Record exposure by 1.0045 to 1.6421 8719.562: Frame time for constraint is 0.1598 8719.578: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8722.875: recentlyReversed: dir 1 ang 17.999795 revtilt 20.994428 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.016 0.077 um, 1.1 5.5 % Error in Z Prediction -0.214 Predicted X = -0.045 StdErr = 0.018 nFit = 3 nDrop = 0 Predicted Y = -0.451 StdErr = 0.028 nFit = 5 nDrop = 0 Predicted Z = 1.795 StdErr = 0.134 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.006, 0.004 um; position -0.039, -0.455 Measured defocus = -0.10 microns changed by 0.00 to target Tracking image shifted -0.001, 0.000 um; position -0.039, -0.456 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0028_18.0.tif Frame alignment results: distance raw = 5.0 smoothed = 4.0 Weighted resid mean = 0.05 max max = 0.10 Mean unweighted max = 0.20 Record image shifted -0.000, 0.012 um Tracking reference shifted by -1.5,-9.0 pixels, -0.000,0.012 um Saved Z = 28 at 13.87 min Tilt = 18.00 Mean = 44 Exp = 1.642 X/Y/Z = -0.038 -0.468 1.797 8750.500: Saved state 28 at 18.0: def 1.697 IS 0.255 -0.393 int 0.12073 exp 1.642 8750.594: Restored state 26 at -38.0 for tiltInd 29 def 4.556 IS -0.074 0.281 8750.594: SetIntensity called with factor 1.0049 8750.609: Changing (ideal) Record exposure by 1.0049 to 1.6504 8750.625: Frame time for constraint is 0.1598 8750.625: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.007 -0.017 um, 0.5 1.2 % Error in Z Prediction 0.002 Predicted X = -0.068 StdErr = 0.026 nFit = 4 nDrop = 0 Predicted Y = 0.230 StdErr = 0.016 nFit = 6 nDrop = 0 Predicted Z = 4.694 StdErr = 0.177 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.015, -0.092 um; position -0.084, 0.322 Measured defocus = 0.17 microns changed by -0.27 to target Tracking image shifted 0.002, 0.013 um; position -0.085, 0.310 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0029_-40.0.tif Frame alignment results: distance raw = 7.4 smoothed = 6.1 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.19 Record image shifted -0.002, -0.009 um Tracking reference shifted by 2.5,6.1 pixels, -0.002,-0.009 um Saved Z = 29 at 14.43 min Tilt = -39.99 Mean = 47 Exp = 1.650 X/Y/Z = -0.084 0.318 4.413 8784.015: Saved state 29 at -40.0: def 4.313 IS -0.080 0.326 int 0.12073 exp 1.650 8784.031: SetIntensity called with factor 1.0053 8784.031: Changing (ideal) Record exposure by 1.0053 to 1.6591 8784.047: Frame time for constraint is 0.1598 8784.047: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.015 0.088 um, 1.1 6.4 % Error in Z Prediction -0.281 Predicted X = -0.089 StdErr = 0.029 nFit = 4 nDrop = 0 Predicted Y = 0.307 StdErr = 0.039 nFit = 6 nDrop = 0 Predicted Z = 4.649 StdErr = 0.239 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.007, -0.069 um; position -0.082, 0.375 Measured defocus = -0.08 microns changed by -0.02 to target Tracking image shifted 0.001, 0.006 um; position -0.083, 0.370 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0030_-42.0.tif Frame alignment results: distance raw = 9.8 smoothed = 9.4 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.22 Record image shifted 0.001, -0.003 um Tracking reference shifted by -0.1,2.4 pixels, 0.001,-0.003 um Saved Z = 30 at 14.92 min Tilt = -41.99 Mean = 47 Exp = 1.659 X/Y/Z = -0.084 0.373 4.625 8813.187: Saved state 30 at -42.0: def 4.525 IS -0.106 0.375 int 0.12073 exp 1.659 8813.265: Restored state 28 at 18.0 for tiltInd 31 def 1.697 IS 0.255 -0.393 8813.281: SetIntensity called with factor 1.0049 8813.297: Changing (ideal) Record exposure by 1.0049 to 1.6501 8813.312: Frame time for constraint is 0.1598 8813.312: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8821.156: recentlyReversed: dir 1 ang 19.998050 revtilt 22.995183 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.006 0.066 um, 0.4 4.8 % Error in Z Prediction -0.024 Predicted X = -0.041 StdErr = 0.031 nFit = 4 nDrop = 0 Predicted Y = -0.448 StdErr = 0.021 nFit = 6 nDrop = 0 Predicted Z = 1.631 StdErr = 0.124 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.021, -0.025 um; position -0.062, -0.423 Measured defocus = -0.27 microns changed by 0.17 to target Tracking image shifted -0.003, 0.006 um; position -0.058, -0.429 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0031_20.0.tif Frame alignment results: distance raw = 8.7 smoothed = 8.2 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.22 Record image shifted -0.004, 0.013 um Tracking reference shifted by 0.9,-9.7 pixels, -0.004,0.013 um Saved Z = 31 at 15.52 min Tilt = 20.00 Mean = 44 Exp = 1.650 X/Y/Z = -0.055 -0.441 1.804 8849.078: Saved state 31 at 20.0: def 1.704 IS 0.256 -0.360 int 0.12073 exp 1.650 8849.078: SetIntensity called with factor 1.0053 8849.094: Changing (ideal) Record exposure by 1.0053 to 1.6587 8849.109: Frame time for constraint is 0.1598 8849.109: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8852.344: recentlyReversed: dir 1 ang 21.999305 revtilt 24.998937 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.014 0.006 um, 1.0 0.5 % Error in Z Prediction 0.173 Predicted X = -0.054 StdErr = 0.019 nFit = 3 nDrop = 0 Predicted Y = -0.398 StdErr = 0.030 nFit = 5 nDrop = 0 Predicted Z = 1.575 StdErr = 0.135 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.013, 0.099 um; position -0.040, -0.497 Measured defocus = -0.44 microns changed by 0.34 to target Tracking image shifted -0.001, 0.002 um; position -0.040, -0.500 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0032_22.0.tif Frame alignment results: distance raw = 4.1 smoothed = 3.2 Weighted resid mean = 0.10 max max = 0.16 Mean unweighted max = 0.30 Record image shifted -0.002, 0.016 um Tracking reference shifted by -0.7,-11.7 pixels, -0.002,0.016 um Saved Z = 32 at 16.03 min Tilt = 22.00 Mean = 43 Exp = 1.659 X/Y/Z = -0.038 -0.515 1.916 8879.875: Saved state 32 at 22.0: def 1.816 IS 0.277 -0.435 int 0.12073 exp 1.659 8879.953: Restored state 30 at -42.0 for tiltInd 33 def 4.525 IS -0.106 0.375 8879.969: SetIntensity called with factor 1.0057 8879.984: Changing (ideal) Record exposure by 1.0057 to 1.6686 8879.984: Frame time for constraint is 0.1598 8880.000: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.016 -0.117 um, 1.2 8.5 % Error in Z Prediction 0.341 Predicted X = -0.097 StdErr = 0.026 nFit = 4 nDrop = 0 Predicted Y = 0.450 StdErr = 0.023 nFit = 5 nDrop = 1 Predicted Z = 4.726 StdErr = 0.185 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.003, 0.010 um; position -0.100, 0.440 Measured defocus = -0.31 microns changed by 0.21 to target Tracking image shifted 0.000, -0.001 um; position -0.100, 0.440 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0033_-44.0.tif Frame alignment results: distance raw = 5.7 smoothed = 4.9 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.21 Record image shifted -0.000, 0.004 um Tracking reference shifted by -0.6,-3.1 pixels, -0.000,0.004 um Saved Z = 33 at 16.60 min Tilt = -43.98 Mean = 47 Exp = 1.669 X/Y/Z = -0.100 0.436 4.939 8913.828: Saved state 33 at -44.0: def 4.839 IS -0.122 0.439 int 0.12073 exp 1.669 8913.844: SetIntensity called with factor 1.0062 8913.859: Changing (ideal) Record exposure by 1.0062 to 1.6789 8913.875: Frame time for constraint is 0.1598 8913.875: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.003 -0.014 um, 0.2 1.0 % Error in Z Prediction 0.213 Predicted X = -0.108 StdErr = 0.021 nFit = 4 nDrop = 0 Predicted Y = 0.530 StdErr = 0.018 nFit = 6 nDrop = 0 Predicted Z = 4.932 StdErr = 0.230 nFit = 5 nDrop = 0 Need autofocus Measured defocus = 0.21 microns changed by -0.31 to target 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0034_-46.0.tif Frame alignment results: distance raw = 16.2 smoothed = 15.0 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.34 Record image shifted -0.007, 0.071 um Saved Z = 34 at 17.02 min Tilt = -45.99 Mean = 47 Exp = 1.679 X/Y/Z = -0.101 0.459 4.621 8939.515: Saved state 34 at -46.0: def 4.521 IS -0.132 0.460 int 0.12073 exp 1.679 8939.594: Restored state 32 at 22.0 for tiltInd 35 def 1.816 IS 0.277 -0.435 8939.609: SetIntensity called with factor 1.0057 8939.625: Changing (ideal) Record exposure by 1.0057 to 1.6682 8939.625: Frame time for constraint is 0.1598 8939.640: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 8947.422: recentlyReversed: dir 1 ang 23.999060 revtilt 26.994193 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.007 -0.071 um, 0.5 5.1 % Error in Z Prediction -0.311 Predicted X = -0.043 StdErr = 0.025 nFit = 3 nDrop = 0 Predicted Y = -0.509 StdErr = 0.062 nFit = 6 nDrop = 0 Predicted Z = 1.871 StdErr = 0.116 nFit = 4 nDrop = 1 Need regular tracking - Need autofocus Tracking image shifted 0.009, 0.034 um; position -0.052, -0.543 Measured defocus = -0.21 microns changed by 0.11 to target Tracking image shifted -0.004, 0.008 um; position -0.048, -0.551 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0035_24.0.tif Frame alignment results: distance raw = 6.7 smoothed = 6.1 Weighted resid mean = 0.11 max max = 0.18 Mean unweighted max = 0.24 Record image shifted -0.004, 0.012 um Tracking reference shifted by 1.1,-9.3 pixels, -0.004,0.012 um Saved Z = 35 at 17.62 min Tilt = 24.00 Mean = 43 Exp = 1.668 X/Y/Z = -0.045 -0.563 1.985 8975.109: Saved state 35 at 24.0: def 1.885 IS 0.305 -0.473 int 0.12073 exp 1.668 8975.125: SetIntensity called with factor 1.0061 8975.140: Changing (ideal) Record exposure by 1.0061 to 1.6784 8975.156: Frame time for constraint is 0.1598 8975.156: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 8978.344: recentlyReversed: dir 1 ang 25.999315 revtilt 28.986449 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.002 -0.054 um, 0.1 3.9 % Error in Z Prediction 0.114 Predicted X = -0.033 StdErr = 0.038 nFit = 4 nDrop = 0 Predicted Y = -0.651 StdErr = 0.055 nFit = 5 nDrop = 0 Predicted Z = 2.044 StdErr = 0.051 nFit = 4 nDrop = 2 Need regular tracking - Need autofocus Tracking image shifted -0.002, -0.030 um; position -0.031, -0.621 Measured defocus = -0.22 microns changed by 0.12 to target Tracking image shifted -0.002, 0.002 um; position -0.029, -0.622 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0036_26.0.tif Frame alignment results: distance raw = 6.3 smoothed = 5.4 Weighted resid mean = 0.09 max max = 0.14 Mean unweighted max = 0.31 Record image shifted -0.001, 0.017 um Tracking reference shifted by -1.6,-12.6 pixels, -0.001,0.017 um Saved Z = 36 at 18.14 min Tilt = 26.00 Mean = 42 Exp = 1.678 X/Y/Z = -0.028 -0.639 2.165 9006.640: Saved state 36 at 26.0: def 2.065 IS 0.328 -0.550 int 0.12073 exp 1.678 9006.719: Restored state 34 at -46.0 for tiltInd 37 def 4.521 IS -0.132 0.460 9006.734: SetIntensity called with factor 1.0066 9006.750: Changing (ideal) Record exposure by 1.0066 to 1.6901 9006.765: Frame time for constraint is 0.1598 9006.781: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.004 0.012 um, 0.3 0.8 % Error in Z Prediction 0.121 Predicted X = -0.110 StdErr = 0.021 nFit = 4 nDrop = 0 Predicted Y = 0.503 StdErr = 0.030 nFit = 5 nDrop = 0 Predicted Z = 4.860 StdErr = 0.243 nFit = 6 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.013, 0.052 um; position -0.124, 0.451 Measured defocus = -0.09 microns changed by -0.01 to target Tracking image shifted 0.000, 0.007 um; position -0.124, 0.444 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0037_-48.0.tif Frame alignment results: distance raw = 7.1 smoothed = 5.9 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.30 Record image shifted -0.002, -0.003 um Tracking reference shifted by 2.0,2.2 pixels, -0.002,-0.003 um Saved Z = 37 at 18.73 min Tilt = -47.99 Mean = 46 Exp = 1.690 X/Y/Z = -0.122 0.448 4.837 9041.922: Saved state 37 at -48.0: def 4.737 IS -0.109 0.461 int 0.12073 exp 1.690 9041.937: SetIntensity called with factor 1.0071 9041.953: Changing (ideal) Record exposure by 1.0071 to 1.7021 9041.969: Frame time for constraint is 0.1598 9041.984: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.012 -0.056 um, 0.8 4.0 % Error in Z Prediction -0.022 Predicted X = -0.136 StdErr = 0.023 nFit = 4 nDrop = 0 Predicted Y = 0.424 StdErr = 0.025 nFit = 5 nDrop = 1 Predicted Z = 4.866 StdErr = 0.238 nFit = 6 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.023, 0.016 um; position -0.112, 0.408 Measured defocus = -0.22 microns changed by 0.12 to target Tracking image shifted -0.000, 0.008 um; position -0.112, 0.400 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0038_-50.0.tif Frame alignment results: distance raw = 11.7 smoothed = 10.7 Weighted resid mean = 0.09 max max = 0.15 Mean unweighted max = 0.33 Record image shifted 0.001, -0.003 um Tracking reference shifted by -0.1,2.3 pixels, 0.001,-0.003 um Saved Z = 38 at 19.22 min Tilt = -49.98 Mean = 45 Exp = 1.702 X/Y/Z = -0.113 0.403 4.981 9071.078: Saved state 38 at -50.0: def 4.881 IS -0.095 0.417 int 0.12073 exp 1.702 9071.156: Restored state 36 at 26.0 for tiltInd 39 def 2.065 IS 0.328 -0.550 9071.172: SetIntensity called with factor 1.0066 9071.187: Changing (ideal) Record exposure by 1.0066 to 1.6895 9071.203: Frame time for constraint is 0.1598 9071.219: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 9079.312: recentlyReversed: dir 1 ang 27.999070 revtilt 30.998202 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.023 -0.021 um, 1.6 1.5 % Error in Z Prediction 0.115 Predicted X = -0.027 StdErr = 0.018 nFit = 3 nDrop = 0 Predicted Y = -0.755 StdErr = 0.053 nFit = 5 nDrop = 0 Predicted Z = 2.256 StdErr = 0.054 nFit = 4 nDrop = 2 Need regular tracking - Tracking image shifted 0.037, -0.108 um; position -0.064, -0.646 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0039_28.0.tif Frame alignment results: distance raw = 10.8 smoothed = 9.8 Weighted resid mean = 0.11 max max = 0.19 Mean unweighted max = 0.32 Record image shifted -0.007, 0.028 um Tracking reference shifted by 1.4,-21.0 pixels, -0.007,0.028 um Saved Z = 39 at 19.73 min Tilt = 28.00 Mean = 42 Exp = 1.689 X/Y/Z = -0.057 -0.674 2.256 9102.031: Saved state 39 at 28.0: def 2.156 IS 0.368 -0.565 int 0.12073 exp 1.689 9102.047: SetIntensity called with factor 1.0071 9102.047: Changing (ideal) Record exposure by 1.0071 to 1.7015 9102.062: Frame time for constraint is 0.1598 9102.078: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 9105.265: recentlyReversed: dir 1 ang 29.999325 revtilt 32.997957 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.029 0.081 um, 2.1 5.8 % Predicted X = -0.055 StdErr = 0.034 nFit = 3 nDrop = 0 Predicted Y = -0.718 StdErr = 0.026 nFit = 5 nDrop = 1 Predicted Z = 2.371 StdErr = 0.066 nFit = 4 nDrop = 2 Need regular tracking - Need autofocus Tracking image shifted -0.012, -0.003 um; position -0.044, -0.715 Measured defocus = 0.25 microns changed by -0.35 to target Tracking image shifted -0.003, 0.010 um; position -0.041, -0.725 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0040_30.0.tif Frame alignment results: distance raw = 5.7 smoothed = 4.1 Weighted resid mean = 0.09 max max = 0.14 Mean unweighted max = 0.37 Record image shifted 0.000, 0.012 um Tracking reference shifted by -1.7,-8.3 pixels, 0.000,0.012 um Saved Z = 40 at 20.26 min Tilt = 30.00 Mean = 41 Exp = 1.701 X/Y/Z = -0.041 -0.736 2.028 9133.672: Saved state 40 at 30.0: def 1.928 IS 0.384 -0.628 int 0.12073 exp 1.701 9133.750: Restored state 38 at -50.0 for tiltInd 41 def 4.881 IS -0.095 0.417 9133.765: SetIntensity called with factor 1.0077 9133.781: Changing (ideal) Record exposure by 1.0077 to 1.7152 9133.797: Frame time for constraint is 0.1598 9133.797: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.014 -0.018 um, 1.0 1.3 % Error in Z Prediction -0.343 Predicted X = -0.124 StdErr = 0.022 nFit = 3 nDrop = 0 Predicted Y = 0.320 StdErr = 0.004 nFit = 5 nDrop = 1 Predicted Z = 4.985 StdErr = 0.210 nFit = 7 nDrop = 0 Need autofocus Measured defocus = -0.48 microns changed by 0.38 to target 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0041_-52.0.tif Frame alignment results: distance raw = 8.2 smoothed = 6.4 Weighted resid mean = 0.09 max max = 0.14 Mean unweighted max = 0.51 Trimming A by 36 36, reference by 21 26 Record image shifted 0.020, -0.063 um Saved Z = 41 at 20.81 min Tilt = -51.99 Mean = 44 Exp = 1.715 X/Y/Z = -0.144 0.383 5.362 9166.969: Saved state 41 at -52.0: def 5.262 IS -0.059 0.416 int 0.12073 exp 1.715 9166.984: SetIntensity called with factor 1.0082 9167.000: Changing (ideal) Record exposure by 1.0082 to 1.7293 9167.031: Frame time for constraint is 0.1598 9167.031: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.020 0.063 um, 1.5 4.6 % Error in Z Prediction 0.377 Predicted X = -0.163 StdErr = 0.046 nFit = 4 nDrop = 0 Predicted Y = 0.297 StdErr = 0.032 nFit = 6 nDrop = 0 Predicted Z = 5.542 StdErr = 0.120 nFit = 4 nDrop = 3 Need regular tracking - Need autofocus Tracking image shifted -0.034, -0.028 um; position -0.129, 0.325 Measured defocus = -1.07 microns changed by 0.97 to target Tracking image shifted 0.013, -0.060 um; position -0.142, 0.385 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0042_-54.0.tif Frame alignment results: distance raw = 9.6 smoothed = 8.6 Weighted resid mean = 0.11 max max = 0.20 Mean unweighted max = 0.63 Trimming A by 23 56, reference by 36 41 Record image shifted -0.001, 0.006 um Tracking reference shifted by -0.1,-4.4 pixels, -0.001,0.006 um Saved Z = 42 at 21.30 min Tilt = -53.98 Mean = 44 Exp = 1.729 X/Y/Z = -0.141 0.379 6.599 9196.359: Saved state 42 at -54.0: def 6.499 IS -0.060 0.411 int 0.12073 exp 1.729 9196.453: Restored state 40 at 30.0 for tiltInd 43 def 1.928 IS 0.384 -0.628 9196.469: SetIntensity called with factor 1.0076 9196.484: Changing (ideal) Record exposure by 1.0076 to 1.7144 9196.500: Frame time for constraint is 0.1598 9196.500: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 9204.922: recentlyReversed: dir 1 ang 31.999080 revtilt 34.999212 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.022 0.082 um, 1.6 5.9 % Error in Z Prediction 1.057 Predicted X = -0.055 StdErr = 0.033 nFit = 3 nDrop = 0 Predicted Y = -0.785 StdErr = 0.028 nFit = 5 nDrop = 0 Predicted Z = 2.178 StdErr = 0.130 nFit = 6 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.013, -0.074 um; position -0.068, -0.711 Measured defocus = 0.16 microns changed by -0.26 to target Tracking image shifted -0.004, 0.013 um; position -0.064, -0.724 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0043_32.0.tif Frame alignment results: distance raw = 5.7 smoothed = 4.4 Weighted resid mean = 0.13 max max = 0.20 Mean unweighted max = 0.30 Record image shifted -0.006, 0.017 um Tracking reference shifted by 1.7,-13.4 pixels, -0.006,0.017 um Saved Z = 43 at 21.92 min Tilt = 32.00 Mean = 41 Exp = 1.714 X/Y/Z = -0.058 -0.742 1.927 9233.047: Saved state 43 at 32.0: def 1.827 IS 0.402 -0.624 int 0.12073 exp 1.714 9233.062: SetIntensity called with factor 1.0082 9233.078: Changing (ideal) Record exposure by 1.0082 to 1.7285 9233.094: Frame time for constraint is 0.1598 9233.109: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 9236.281: recentlyReversed: dir 1 ang 33.999335 revtilt 36.997467 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.003 0.043 um, 0.2 3.1 % Error in Z Prediction -0.251 Predicted X = -0.053 StdErr = 0.025 nFit = 3 nDrop = 0 Predicted Y = -0.750 StdErr = 0.032 nFit = 5 nDrop = 0 Predicted Z = 2.007 StdErr = 0.163 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.008, 0.020 um; position -0.045, -0.770 Measured defocus = -0.13 microns changed by 0.03 to target Tracking image shifted -0.002, 0.005 um; position -0.043, -0.776 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0044_34.0.tif Frame alignment results: distance raw = 7.3 smoothed = 6.6 Weighted resid mean = 0.13 max max = 0.20 Mean unweighted max = 0.49 Record image shifted -0.001, 0.019 um Tracking reference shifted by -1.5,-13.7 pixels, -0.001,0.019 um Saved Z = 44 at 22.44 min Tilt = 34.00 Mean = 40 Exp = 1.728 X/Y/Z = -0.042 -0.794 2.040 9264.328: Saved state 44 at 34.0: def 1.940 IS 0.413 -0.679 int 0.12073 exp 1.728 9264.422: Restored state 42 at -54.0 for tiltInd 45 def 6.499 IS -0.060 0.411 9264.437: SetIntensity called with factor 1.0088 9264.453: Changing (ideal) Record exposure by 1.0088 to 1.7446 9264.453: Frame time for constraint is 0.1598 9264.469: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.011 -0.044 um, 0.8 3.2 % Error in Z Prediction 0.033 Predicted X = -0.160 StdErr = 0.049 nFit = 4 nDrop = 0 Predicted Y = 0.367 StdErr = 0.029 nFit = 5 nDrop = 0 Predicted Z = 6.156 StdErr = 0.601 nFit = 7 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.005, -0.063 um; position -0.165, 0.430 Measured defocus = -0.51 microns changed by 0.41 to target Trimming A by 25 25, reference by 20 20 Tracking image shifted 0.000, 0.015 um; position -0.166, 0.415 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0045_-56.0.tif Frame alignment results: distance raw = 8.8 smoothed = 7.1 Weighted resid mean = 0.15 max max = 0.24 Mean unweighted max = 0.66 Trimming A by 33 66, reference by 46 51 Record image shifted -0.002, 0.004 um Tracking reference shifted by 0.6,-3.2 pixels, -0.002,0.004 um Saved Z = 45 at 23.03 min Tilt = -55.99 Mean = 42 Exp = 1.745 X/Y/Z = -0.164 0.411 6.542 9299.844: Saved state 45 at -56.0: def 6.442 IS -0.055 0.452 int 0.12073 exp 1.745 9299.859: SetIntensity called with factor 1.0095 9299.875: Changing (ideal) Record exposure by 1.0095 to 1.7611 9299.890: Frame time for constraint is 0.1598 9299.906: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.004 0.044 um, 0.3 3.2 % Error in Z Prediction 0.387 Predicted X = -0.168 StdErr = 0.039 nFit = 4 nDrop = 0 Predicted Y = 0.460 StdErr = 0.008 nFit = 5 nDrop = 1 Predicted Z = 7.016 StdErr = 0.464 nFit = 6 nDrop = 0 Need regular tracking - Need autofocus Trimming A by 20 30, reference by 25 35 Tracking image shifted -0.006, 0.029 um; position -0.163, 0.431 Measured defocus = -0.36 microns changed by 0.26 to target Trimming A by 22 35, reference by 25 35 Tracking image shifted -0.001, -0.003 um; position -0.162, 0.433 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0046_-58.0.tif Frame alignment results: distance raw = 7.5 smoothed = 6.4 Weighted resid mean = 0.26 max max = 0.38 Mean unweighted max = 0.95 Trimming A by 33 76, reference by 40 71 Record image shifted 0.001, 0.006 um Tracking reference shifted by -1.6,-4.1 pixels, 0.001,0.006 um Saved Z = 46 at 23.51 min Tilt = -57.98 Mean = 41 Exp = 1.761 X/Y/Z = -0.163 0.427 7.284 9328.969: Saved state 46 at -58.0: def 7.184 IS -0.064 0.466 int 0.12073 exp 1.761 9329.047: Restored state 44 at 34.0 for tiltInd 47 def 1.940 IS 0.413 -0.679 9329.062: SetIntensity called with factor 1.0088 9329.078: Changing (ideal) Record exposure by 1.0088 to 1.7436 9329.094: Frame time for constraint is 0.1598 9329.109: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 9337.672: recentlyReversed: dir 1 ang 35.999090 revtilt 38.998222 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.005 -0.033 um, 0.4 2.4 % Error in Z Prediction 0.268 Predicted X = -0.048 StdErr = 0.025 nFit = 3 nDrop = 0 Predicted Y = -0.820 StdErr = 0.039 nFit = 5 nDrop = 0 Predicted Z = 1.986 StdErr = 0.133 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.022, -0.052 um; position -0.070, -0.768 Measured defocus = 0.01 microns changed by -0.11 to target Tracking image shifted -0.005, 0.012 um; position -0.065, -0.780 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0047_36.0.tif Frame alignment results: distance raw = 9.0 smoothed = 8.5 Weighted resid mean = 0.17 max max = 0.26 Mean unweighted max = 0.47 Record image shifted -0.006, 0.019 um Tracking reference shifted by 1.7,-14.4 pixels, -0.006,0.019 um Saved Z = 47 at 24.13 min Tilt = 36.00 Mean = 39 Exp = 1.744 X/Y/Z = -0.059 -0.798 1.886 9365.719: Saved state 47 at 36.0: def 1.786 IS 0.429 -0.673 int 0.12073 exp 1.744 9365.734: SetIntensity called with factor 1.0094 9365.750: Changing (ideal) Record exposure by 1.0094 to 1.7600 9365.765: Frame time for constraint is 0.1598 9365.781: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 9368.984: recentlyReversed: dir 1 ang 37.999345 revtilt 40.993978 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.011 0.022 um, 0.8 1.6 % Error in Z Prediction -0.100 Predicted X = -0.054 StdErr = 0.025 nFit = 3 nDrop = 0 Predicted Y = -0.806 StdErr = 0.040 nFit = 5 nDrop = 0 Predicted Z = 1.858 StdErr = 0.095 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.009, 0.033 um; position -0.045, -0.839 Measured defocus = -0.19 microns changed by 0.09 to target Tracking image shifted -0.002, 0.006 um; position -0.043, -0.845 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0048_38.0.tif Frame alignment results: distance raw = 12.8 smoothed = 12.7 Weighted resid mean = 0.17 max max = 0.28 Mean unweighted max = 0.94 Record image shifted -0.002, 0.020 um Tracking reference shifted by -1.0,-14.8 pixels, -0.002,0.020 um Saved Z = 48 at 24.65 min Tilt = 38.00 Mean = 38 Exp = 1.760 X/Y/Z = -0.040 -0.865 1.948 9396.969: Saved state 48 at 38.0: def 1.848 IS 0.445 -0.742 int 0.12073 exp 1.760 9397.047: Restored state 46 at -58.0 for tiltInd 49 def 7.184 IS -0.064 0.466 9397.062: SetIntensity called with factor 1.0102 9397.078: Changing (ideal) Record exposure by 1.0102 to 1.7790 9397.094: Frame time for constraint is 0.1598 9397.109: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.014 -0.059 um, 1.0 4.2 % Error in Z Prediction 0.090 Predicted X = -0.179 StdErr = 0.018 nFit = 3 nDrop = 0 Predicted Y = 0.486 StdErr = 0.017 nFit = 6 nDrop = 0 Predicted Z = 7.607 StdErr = 0.414 nFit = 7 nDrop = 0 Need regular tracking - Need autofocus Trimming A by 22 35, reference by 30 40 Tracking image shifted 0.008, 0.011 um; position -0.187, 0.474 Measured defocus = 0.12 microns changed by -0.22 to target Trimming A by 25 30, reference by 30 40 Tracking image shifted -0.000, 0.008 um; position -0.186, 0.466 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0049_-60.0.tif Frame alignment results: distance raw = 13.2 smoothed = 13.1 Weighted resid mean = 0.27 max max = 0.46 Mean unweighted max = 1.26 Trimming A by 61 61, reference by 34 81 Record image shifted -0.003, 0.017 um Tracking reference shifted by -0.4,-12.6 pixels, -0.003,0.017 um Saved Z = 49 at 25.25 min Tilt = -59.99 Mean = 39 Exp = 1.779 X/Y/Z = -0.184 0.449 7.371 9432.906: Saved state 49 at -60.0: def 7.271 IS -0.057 0.496 int 0.12073 exp 1.779 9432.922: SetIntensity called with factor 1.0109 9432.937: Changing (ideal) Record exposure by 1.0109 to 1.7984 9432.953: Frame time for constraint is 0.1598 9432.969: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.005 -0.036 um, 0.4 2.6 % Error in Z Prediction -0.236 Predicted X = -0.190 StdErr = 0.016 nFit = 3 nDrop = 0 Predicted Y = 0.482 StdErr = 0.022 nFit = 5 nDrop = 0 Predicted Z = 8.052 StdErr = 0.389 nFit = 7 nDrop = 0 Need regular tracking - Need autofocus Trimming A by 20 20, reference by 30 40 Tracking image shifted -0.015, 0.095 um; position -0.175, 0.387 Measured defocus = 0.64 microns changed by -0.74 to target Trimming A by 25 30, reference by 30 40 Tracking image shifted -0.000, -0.006 um; position -0.174, 0.394 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0050_-62.0.tif Frame alignment results: distance raw = 10.9 smoothed = 9.3 Weighted resid mean = 0.58 max max = 0.98 Mean unweighted max = 3.50 Trimming A by 23 76, reference by 21 86 Record image shifted -0.002, 0.006 um Tracking reference shifted by 0.7,-4.9 pixels, -0.002,0.006 um Saved Z = 50 at 25.74 min Tilt = -61.99 Mean = 36 Exp = 1.798 X/Y/Z = -0.172 0.387 7.322 9462.578: Saved state 50 at -62.0: def 7.222 IS -0.037 0.435 int 0.12073 exp 1.798 9462.672: Restored state 48 at 38.0 for tiltInd 51 def 1.848 IS 0.445 -0.742 9462.687: SetIntensity called with factor 1.0101 9462.703: Changing (ideal) Record exposure by 1.0101 to 1.7778 9462.719: Frame time for constraint is 0.1598 9462.734: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 9472.172: recentlyReversed: dir 1 ang 39.999600 revtilt 42.998732 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.017 -0.095 um, 1.3 6.9 % Error in Z Prediction -0.730 Predicted X = -0.046 StdErr = 0.027 nFit = 3 nDrop = 0 Predicted Y = -0.918 StdErr = 0.042 nFit = 5 nDrop = 0 Predicted Z = 1.905 StdErr = 0.097 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.020, -0.077 um; position -0.065, -0.841 Using peak at -6.6,0.8, rejecting 2.36 times higher peak at 0.0,-0.1 Measured defocus = -0.10 microns changed by -0.00 to target Tracking image shifted -0.004, 0.008 um; position -0.061, -0.849 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0051_40.0.tif Frame alignment results: distance raw = 14.4 smoothed = 14.3 Weighted resid mean = 0.19 max max = 0.33 Mean unweighted max = 0.86 Record image shifted -0.006, 0.016 um Tracking reference shifted by 2.1,-12.2 pixels, -0.006,0.016 um Saved Z = 51 at 26.37 min Tilt = 40.00 Mean = 37 Exp = 1.778 X/Y/Z = -0.055 -0.865 1.909 9500.219: Saved state 51 at 40.0: def 1.809 IS 0.458 -0.734 int 0.12073 exp 1.778 9500.234: SetIntensity called with factor 1.0108 9500.250: Changing (ideal) Record exposure by 1.0108 to 1.7971 9500.265: Frame time for constraint is 0.1598 9500.281: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 9503.515: recentlyReversed: dir 1 ang 41.999355 revtilt 44.998487 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.010 0.053 um, 0.7 3.8 % Error in Z Prediction 0.004 Predicted X = -0.048 StdErr = 0.025 nFit = 3 nDrop = 0 Predicted Y = -0.887 StdErr = 0.049 nFit = 5 nDrop = 0 Predicted Z = 1.888 StdErr = 0.079 nFit = 6 nDrop = 0 Need regular tracking - Tracking image shifted -0.005, 0.012 um; position -0.043, -0.898 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0052_42.0.tif Frame alignment results: distance raw = 7.6 smoothed = 7.0 Weighted resid mean = 0.18 max max = 0.31 Mean unweighted max = 0.81 Record image shifted -0.004, 0.029 um Tracking reference shifted by -0.7,-21.5 pixels, -0.004,0.029 um Saved Z = 52 at 26.81 min Tilt = 42.00 Mean = 36 Exp = 1.797 X/Y/Z = -0.038 -0.927 1.888 9526.375: Saved state 52 at 42.0: def 1.788 IS 0.473 -0.799 int 0.12073 exp 1.797 9526.453: Restored state 50 at -62.0 for tiltInd 53 def 7.222 IS -0.037 0.435 9526.469: SetIntensity called with factor 1.0117 9526.484: Changing (ideal) Record exposure by 1.0117 to 1.8195 9526.500: Frame time for constraint is 0.1598 9526.531: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.009 -0.041 um, 0.7 2.9 % Predicted X = -0.182 StdErr = 0.023 nFit = 3 nDrop = 0 Predicted Y = 0.336 StdErr = 0.042 nFit = 5 nDrop = 0 Predicted Z = 7.706 StdErr = 0.347 nFit = 5 nDrop = 1 Need regular tracking - Need autofocus Trimming A by 25 25, reference by 27 40 Tracking image shifted -0.004, 0.001 um; position -0.178, 0.335 Measured defocus = 0.52 microns changed by -0.62 to target Trimming A by 25 25, reference by 27 40 Tracking image shifted -0.002, 0.005 um; position -0.176, 0.330 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0053_-64.0.tif Frame alignment results: distance raw = 26.1 smoothed = 22.1 Weighted resid mean = 1.11 max max = 3.50 Mean unweighted max = 16.20 Trimming A by 76 81, reference by 31 31 Record image shifted -0.003, -0.014 um Tracking reference shifted by 4.1,9.5 pixels, -0.003,-0.014 um Saved Z = 53 at 27.41 min Tilt = -63.99 Mean = 26 Exp = 1.820 X/Y/Z = -0.173 0.344 7.078 9562.406: Saved state 53 at -64.0: def 6.978 IS -0.016 0.397 int 0.12073 exp 1.820 9562.422: SetIntensity called with factor 1.0126 9562.437: Changing (ideal) Record exposure by 1.0126 to 1.8425 9562.453: Frame time for constraint is 0.1598 9562.469: Will be constrained to 1.9174 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.009 0.008 um, 0.7 0.6 % Error in Z Prediction -0.628 Predicted X = -0.153 StdErr = 0.029 nFit = 4 nDrop = 0 Predicted Y = 0.262 StdErr = 0.030 nFit = 6 nDrop = 0 Predicted Z = 7.097 StdErr = 0.185 nFit = 4 nDrop = 2 Need regular tracking - Need autofocus Trimming A by 20 20, reference by 30 35 Tracking image shifted 0.003, 0.016 um; position -0.156, 0.246 Measured defocus = -25.72 microns changed by 16.00 Measured defocus = -32.99 microns WARNING: Autofocus aborted after 2 iterations and focus returned to starting value (7.00) because of inconsistent behavior. Check focus calibration and/or scope alignment if images and specimen look OK Autofocus has failed to behave consistently after several iterations ONE SIDE OF THE SERIES IS BEING ENDED; GOING ON TO FINISH THE OTHER SIDE BASED ON TILT SERIES POLICY SETTINGS 9580.531: Restored state 52 at 42.0 for tiltInd 55 def 1.788 IS 0.473 -0.799 9580.547: SetIntensity called with factor 1.0117 9580.562: Changing (ideal) Record exposure by 1.0117 to 1.8180 9580.578: Frame time for constraint is 0.1598 9580.594: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 9589.734: recentlyReversed: dir 1 ang 43.999110 revtilt 46.993743 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.020 0.082 um, 1.4 5.9 % Error in Z Prediction -0.019 Need regular tracking - Need autofocus Tracking image shifted -0.001, -0.019 um; position -0.037, -0.908 Measured defocus = 0.46 microns changed by -0.56 to target Tracking image shifted -0.010, 0.001 um; position -0.027, -0.910 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0054_44.0.tif Frame alignment results: distance raw = 29.8 smoothed = 30.1 Weighted resid mean = 0.45 max max = 0.77 Mean unweighted max = 29.02 Record image shifted -0.012, 0.022 um Tracking reference shifted by 5.8,-17.3 pixels, -0.012,0.022 um Saved Z = 54 at 28.29 min Tilt = 44.00 Mean = 34 Exp = 1.818 X/Y/Z = -0.015 -0.931 1.328 9615.594: Saved state 54 at 44.0: def 1.228 IS 0.455 -0.814 int 0.12073 exp 1.818 9615.594: SetIntensity called with factor 1.0125 9615.625: Changing (ideal) Record exposure by 1.0125 to 1.8408 9615.640: Frame time for constraint is 0.1598 9615.640: Will be constrained to 1.9174 (frame 0.1598, constraint 0.1598) 9618.859: recentlyReversed: dir 1 ang 45.999364 revtilt 48.998497 back 3.000000 3mte 0.300000 Need regular tracking - Need autofocus Tracking image shifted -0.022, -0.002 um; position 0.007, -0.929 Measured defocus = -0.13 microns changed by 0.03 to target Tracking image shifted -0.004, -0.000 um; position 0.010, -0.928 12 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0055_46.0.tif Frame alignment results: distance raw = 33.0 smoothed = 32.7 Weighted resid mean = 0.59 max max = 1.13 Mean unweighted max = 94.27 Trimming A by 21 31, reference by 21 21 Record image shifted -0.003, 0.028 um Tracking reference shifted by -1.6,-21.0 pixels, -0.003,0.028 um Saved Z = 55 at 28.82 min Tilt = 46.00 Mean = 33 Exp = 1.841 X/Y/Z = 0.014 -0.956 1.362 9647.078: Saved state 55 at 46.0: def 1.262 IS 0.443 -0.852 int 0.12073 exp 1.841 9647.109: SetIntensity called with factor 1.0135 9647.125: Changing (ideal) Record exposure by 1.0135 to 1.8657 9647.140: Frame time for constraint is 0.1598 9647.156: Will be constrained to 1.9174 (frame 0.1598, constraint 0.1598) 9650.453: recentlyReversed: dir 1 ang 47.999619 revtilt 50.998252 back 3.000000 3mte 0.300000 Predicted X = 0.043 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Y = -0.982 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Z = 1.395 StdErr = 0.000 nFit = 2 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.007, -0.018 um; position 0.036, -0.964 Measured defocus = -0.22 microns changed by 0.12 to target Tracking image shifted -0.002, 0.000 um; position 0.037, -0.964 12 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0056_48.0.tif Frame alignment results: distance raw = 32.3 smoothed = 31.9 Weighted resid mean = 0.99 max max = 1.73 Mean unweighted max = 95.88 Trimming A by 28 41, reference by 26 31 Record image shifted -0.003, 0.031 um Tracking reference shifted by -2.1,-23.1 pixels, -0.003,0.031 um Saved Z = 56 at 29.34 min Tilt = 48.00 Mean = 31 Exp = 1.866 X/Y/Z = 0.040 -0.995 1.516 9678.750: Saved state 56 at 48.0: def 1.416 IS 0.438 -0.901 int 0.12073 exp 1.866 9678.765: SetIntensity called with factor 1.0146 9678.781: Changing (ideal) Record exposure by 1.0146 to 1.8930 9678.797: Frame time for constraint is 0.1598 9678.828: Will be constrained to 1.9174 (frame 0.1598, constraint 0.1598) 9682.375: recentlyReversed: dir 1 ang 49.999374 revtilt 53.000507 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.002 -0.013 um, 0.2 1.0 % Error in Z Prediction 0.120 Predicted X = 0.068 StdErr = 0.002 nFit = 3 nDrop = 0 Predicted Y = -1.025 StdErr = 0.010 nFit = 3 nDrop = 0 Predicted Z = 1.589 StdErr = 0.090 nFit = 3 nDrop = 0 Need regular tracking - Tracking image shifted 0.012, -0.014 um; position 0.057, -1.012 12 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037_0057_50.0.tif Frame alignment results: distance raw = 17.9 smoothed = 16.9 Weighted resid mean = 0.74 max max = 1.54 Mean unweighted max = 7.50 Trimming A by 36 51, reference by 28 41 Record image shifted -0.003, 0.035 um Tracking reference shifted by -2.3,-26.2 pixels, -0.003,0.035 um Saved Z = 57 at 29.78 min Tilt = 50.00 Mean = 29 Exp = 1.893 X/Y/Z = 0.060 -1.047 1.589 2021/11/18 17:39 W:\Zhiyang\Krios\20211117\Y00000Lamella03\20211117_037.mrc 0 58 11520 8184 10936 -10.0 50.0 2.00 REG 29.8 1x1 Continuum_K3 26000 1 0.17 LD-DS 20.0 0.0 Tilt series acquired at item # 29 with label 42 (item at -357.14, 185.33, TS at -357.06, 186.59) ################# 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 5.50 angle 0, subarea offset 0 0