19-Nov-21 22:13:31 Trimming A by 50 55, reference by 50 50 Trimming A by 55 55, reference by 50 50 Trimming A by 60 80, reference by 50 50 Trimming A by 75 75, reference by 50 50 Rough eucentricity: changing Z by -14.67 to -32.37, finished. 112669.047: (Initial) alignment to map 39 (60) 112669.047: Target X, Y: 438.167 -20.541 112669.047: Map matches low dose View; staying in low dose but with exp 0.405 intensity 0.19459 bin 4 Trimming A by 23 85 30 60, reference by 24 84 85 70 Disparity in stage position after aligning to center of map frame: 0.32 0.57 Aligning to target with image shift equivalent to stage shift: 0.32 0.56 112676.594: Map matches low dose Preview; staying in low dose but with exp 0.202 intensity 0.12072 bin 4 112692.109: Realigned to map item itself with image shift of -38.5 -48.4 pixels Disparity in stage position after aligning to map item itself: 0.37 0.53 (align shift 0.06 -0.06) Finished aligning to item in 2 rounds Opened new file W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075.mrc STARTING TILT SERIES WITH AUTOMATIC TERMINATION AFTER ERRORS. 112700.094: Doing reverse tilt to 3.00 at 0.00 Trimming A by 21 74 95 80, reference by 24 90 80 65 Measured defocus = -10.09 microns changed by 9.99 to target Measured defocus = 0.18 microns changed by -0.28 to target 112743.906: SetIntensity called with factor 0.9962 112743.906: Changing (ideal) Record exposure by 0.9962 to 1.5918 112743.906: Frame time for constraint is 0.1598 112743.906: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 112743.906: Skipping reverse tilt: dir -1 start -10.00 revang -7.00 revtilt 3.00 maxtilt 69.90 maxerr 0.10 Need regular tracking - Need autofocus Tracking image shifted -0.007, -0.022 um; position -0.355, -0.343 Measured defocus = -0.02 microns changed by -0.08 to target Tracking image shifted 0.000, -0.001 um; position -0.355, -0.343 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0000_-10.0.tif Frame alignment results: distance raw = 7.6 smoothed = 7.6 Weighted resid mean = 0.05 max max = 0.09 Mean unweighted max = 0.25 Saved Z = 0 at -0.00 min Tilt = -10.00 Mean = 49 Exp = 1.592 X/Y/Z = -0.355 -0.343 4.730 112763.844: Saved state 0 at -10.0: def 4.630 IS 0.466 -0.111 int 0.12073 exp 1.592 112763.859: SetIntensity called with factor 1.0002 112763.859: Changing (ideal) Record exposure by 1.0002 to 1.5920 112763.859: Frame time for constraint is 0.1598 112763.859: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Need regular tracking - Need autofocus Tracking image shifted -0.011, 0.062 um; position -0.344, -0.404 Measured defocus = -0.19 microns changed by 0.09 to target Tracking image shifted 0.001, -0.002 um; position -0.345, -0.402 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0001_-12.0.tif Frame alignment results: distance raw = 4.4 smoothed = 3.8 Weighted resid mean = 0.06 max max = 0.09 Mean unweighted max = 0.13 Record image shifted -0.001, 0.008 um Tracking reference shifted by -0.1,-6.3 pixels, -0.001,0.008 um Saved Z = 1 at 0.48 min Tilt = -12.00 Mean = 49 Exp = 1.592 X/Y/Z = -0.344 -0.411 4.816 112792.640: Saved state 1 at -12.0: def 4.716 IS 0.488 -0.178 int 0.12073 exp 1.592 112792.640: SetIntensity called with factor 1.0005 112792.640: Changing (ideal) Record exposure by 1.0005 to 1.5927 112792.640: Frame time for constraint is 0.1598 112792.656: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Predicted X = -0.332 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Y = -0.479 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Z = 4.901 StdErr = 0.000 nFit = 2 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.008, -0.047 um; position -0.340, -0.432 Measured defocus = -0.17 microns changed by 0.07 to target Tracking image shifted 0.001, -0.002 um; position -0.341, -0.431 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0002_-14.0.tif Frame alignment results: distance raw = 4.4 smoothed = 4.1 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.19 Record image shifted -0.001, 0.008 um Tracking reference shifted by -0.2,-6.1 pixels, -0.001,0.008 um Saved Z = 2 at 0.96 min Tilt = -14.00 Mean = 49 Exp = 1.593 X/Y/Z = -0.339 -0.439 4.969 112821.734: Saved state 2 at -14.0: def 4.869 IS 0.498 -0.205 int 0.12073 exp 1.593 112821.812: Restored state 0 at -10.0 for tiltInd 3 def 4.630 IS 0.466 -0.111 112821.828: SetIntensity called with factor 1.0002 112821.828: Changing (ideal) Record exposure by 1.0002 to 1.5920 112821.828: Frame time for constraint is 0.1598 112821.828: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 112826.219: recentlyReversed: dir 1 ang -7.998520 revtilt -5.000387 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.007 0.040 um, 0.5 2.9 % Error in Z Prediction 0.068 Need regular tracking - Need autofocus Tracking image shifted 0.032, -0.162 um; position -0.387, -0.180 Measured defocus = -0.06 microns changed by -0.04 to target Tracking image shifted 0.001, -0.002 um; position -0.388, -0.178 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0003_-8.0.tif Frame alignment results: distance raw = 7.4 smoothed = 7.3 Weighted resid mean = 0.05 max max = 0.07 Mean unweighted max = 0.23 Record image shifted -0.002, -0.002 um Tracking reference shifted by 2.0,1.1 pixels, -0.002,-0.002 um Saved Z = 3 at 1.49 min Tilt = -8.00 Mean = 49 Exp = 1.592 X/Y/Z = -0.386 -0.176 4.686 112853.703: Saved state 3 at -8.0: def 4.586 IS 0.413 0.052 int 0.12073 exp 1.592 112853.719: SetIntensity called with factor 1.0005 112853.719: Changing (ideal) Record exposure by 1.0005 to 1.5927 112853.719: Frame time for constraint is 0.1598 112853.719: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 112857.140: recentlyReversed: dir 1 ang -5.998265 revtilt -3.004132 back 3.000000 3mte 0.300000 Need regular tracking - Need autofocus Tracking image shifted 0.010, -0.072 um; position -0.396, -0.104 Measured defocus = -0.07 microns changed by -0.03 to target Tracking image shifted 0.001, -0.002 um; position -0.397, -0.102 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0004_-6.0.tif Frame alignment results: distance raw = 8.8 smoothed = 8.5 Weighted resid mean = 0.05 max max = 0.08 Mean unweighted max = 0.19 Record image shifted -0.000, -0.004 um Tracking reference shifted by 0.5,2.5 pixels, -0.000,-0.004 um Saved Z = 4 at 2.01 min Tilt = -6.00 Mean = 49 Exp = 1.593 X/Y/Z = -0.397 -0.099 4.653 112884.719: Saved state 4 at -6.0: def 4.553 IS 0.386 0.127 int 0.12073 exp 1.593 112884.797: Restored state 2 at -14.0 for tiltInd 5 def 4.869 IS 0.498 -0.205 112884.812: SetIntensity called with factor 1.0008 112884.812: Changing (ideal) Record exposure by 1.0008 to 1.5940 112884.812: Frame time for constraint is 0.1598 112884.812: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Predicted X = -0.330 StdErr = 0.006 nFit = 3 nDrop = 0 Predicted Y = -0.494 StdErr = 0.030 nFit = 3 nDrop = 0 Predicted Z = 5.077 StdErr = 0.051 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.027, -0.083 um; position -0.357, -0.410 Measured defocus = -0.10 microns changed by 0.00 to target Tracking image shifted 0.001, -0.002 um; position -0.358, -0.408 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0005_-16.0.tif Frame alignment results: distance raw = 4.3 smoothed = 3.5 Weighted resid mean = 0.05 max max = 0.08 Mean unweighted max = 0.18 Record image shifted -0.003, 0.010 um Tracking reference shifted by 1.0,-7.4 pixels, -0.003,0.010 um Saved Z = 5 at 2.52 min Tilt = -16.00 Mean = 49 Exp = 1.594 X/Y/Z = -0.355 -0.418 5.079 112915.265: Saved state 5 at -16.0: def 4.979 IS 0.501 -0.178 int 0.12073 exp 1.594 112915.265: SetIntensity called with factor 1.0011 112915.265: Changing (ideal) Record exposure by 1.0011 to 1.5957 112915.265: Frame time for constraint is 0.1598 112915.281: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.024 0.076 um, 1.8 5.5 % Error in Z Prediction 0.002 Predicted X = -0.357 StdErr = 0.015 nFit = 3 nDrop = 0 Predicted Y = -0.429 StdErr = 0.036 nFit = 3 nDrop = 0 Predicted Z = 5.218 StdErr = 0.032 nFit = 3 nDrop = 0 Need regular tracking - Tracking image shifted -0.003, -0.032 um; position -0.353, -0.397 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0006_-18.0.tif Frame alignment results: distance raw = 4.7 smoothed = 4.0 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.17 Record image shifted -0.001, 0.007 um Tracking reference shifted by -0.4,-5.2 pixels, -0.001,0.007 um Saved Z = 6 at 2.92 min Tilt = -17.99 Mean = 49 Exp = 1.596 X/Y/Z = -0.353 -0.404 5.218 112939.015: Saved state 6 at -18.0: def 5.118 IS 0.493 -0.167 int 0.12073 exp 1.596 112939.094: Restored state 4 at -6.0 for tiltInd 7 def 4.553 IS 0.386 0.127 112939.094: SetIntensity called with factor 1.0008 112939.094: Changing (ideal) Record exposure by 1.0008 to 1.5940 112939.094: Frame time for constraint is 0.1598 112939.109: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 112943.875: recentlyReversed: dir 1 ang -3.999010 revtilt -1.000877 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.004 0.025 um, 0.3 1.8 % Predicted X = -0.408 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Y = -0.022 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Z = 4.620 StdErr = 0.000 nFit = 2 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.007, 0.025 um; position -0.415, -0.046 Measured defocus = -0.11 microns changed by 0.01 to target Tracking image shifted 0.000, -0.002 um; position -0.416, -0.045 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0007_-4.0.tif Frame alignment results: distance raw = 10.2 smoothed = 10.2 Weighted resid mean = 0.05 max max = 0.08 Mean unweighted max = 0.18 Record image shifted -0.001, 0.000 um Tracking reference shifted by 0.7,-0.2 pixels, -0.001,0.000 um Saved Z = 7 at 3.46 min Tilt = -4.00 Mean = 49 Exp = 1.594 X/Y/Z = -0.415 -0.045 4.635 112971.656: Saved state 7 at -4.0: def 4.535 IS 0.375 0.184 int 0.12073 exp 1.594 112971.656: SetIntensity called with factor 1.0011 112971.672: Changing (ideal) Record exposure by 1.0011 to 1.5957 112971.672: Frame time for constraint is 0.1598 112971.672: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 112975.109: recentlyReversed: dir 1 ang -1.998255 revtilt 0.998378 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.006 -0.023 um, 0.5 1.7 % Error in Z Prediction 0.014 Predicted X = -0.428 StdErr = 0.005 nFit = 3 nDrop = 0 Predicted Y = 0.025 StdErr = 0.017 nFit = 3 nDrop = 0 Predicted Z = 4.607 StdErr = 0.011 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.011, 0.069 um; position -0.417, -0.045 Measured defocus = -0.11 microns changed by 0.01 to target Tracking image shifted 0.001, -0.001 um; position -0.418, -0.043 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0008_-2.0.tif Frame alignment results: distance raw = 13.3 smoothed = 13.4 Weighted resid mean = 0.05 max max = 0.10 Mean unweighted max = 0.24 Record image shifted -0.001, -0.001 um Tracking reference shifted by 0.7,0.9 pixels, -0.001,-0.001 um Saved Z = 8 at 3.98 min Tilt = -2.00 Mean = 48 Exp = 1.596 X/Y/Z = -0.417 -0.042 4.617 113002.687: Saved state 8 at -2.0: def 4.517 IS 0.376 0.188 int 0.12073 exp 1.596 113002.765: Restored state 6 at -18.0 for tiltInd 9 def 5.118 IS 0.493 -0.167 113002.765: SetIntensity called with factor 1.0014 113002.765: Changing (ideal) Record exposure by 1.0014 to 1.5979 113002.781: Frame time for constraint is 0.1598 113002.781: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.011 -0.067 um, 0.8 4.8 % Error in Z Prediction 0.010 Predicted X = -0.362 StdErr = 0.013 nFit = 3 nDrop = 0 Predicted Y = -0.386 StdErr = 0.006 nFit = 3 nDrop = 0 Predicted Z = 5.350 StdErr = 0.043 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.002, 0.017 um; position -0.364, -0.403 Measured defocus = 0.04 microns changed by -0.14 to target Tracking image shifted 0.001, -0.001 um; position -0.365, -0.402 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0009_-20.0.tif Frame alignment results: distance raw = 5.5 smoothed = 5.2 Weighted resid mean = 0.05 max max = 0.09 Mean unweighted max = 0.22 Record image shifted -0.003, 0.010 um Tracking reference shifted by 0.8,-7.4 pixels, -0.003,0.010 um Saved Z = 9 at 4.49 min Tilt = -19.99 Mean = 49 Exp = 1.598 X/Y/Z = -0.362 -0.411 5.206 113033.640: Saved state 9 at -20.0: def 5.106 IS 0.504 -0.168 int 0.12073 exp 1.598 113033.640: SetIntensity called with factor 1.0017 113033.640: Changing (ideal) Record exposure by 1.0017 to 1.6006 113033.656: Frame time for constraint is 0.1598 113033.656: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.000 -0.025 um, 0.0 1.8 % Error in Z Prediction -0.143 Predicted X = -0.364 StdErr = 0.008 nFit = 3 nDrop = 0 Predicted Y = -0.405 StdErr = 0.015 nFit = 3 nDrop = 0 Predicted Z = 5.290 StdErr = 0.042 nFit = 3 nDrop = 0 Need autofocus Measured defocus = -0.10 microns changed by 0.00 to target 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0010_-22.0.tif Frame alignment results: distance raw = 6.8 smoothed = 6.3 Weighted resid mean = 0.06 max max = 0.09 Mean unweighted max = 0.13 Record image shifted -0.012, 0.067 um Saved Z = 10 at 4.92 min Tilt = -21.99 Mean = 49 Exp = 1.601 X/Y/Z = -0.351 -0.472 5.294 113058.984: Saved state 10 at -22.0: def 5.194 IS 0.524 -0.227 int 0.12073 exp 1.601 113059.078: Restored state 8 at -2.0 for tiltInd 11 def 4.517 IS 0.376 0.188 113059.078: SetIntensity called with factor 1.0014 113059.078: Changing (ideal) Record exposure by 1.0014 to 1.5979 113059.094: Frame time for constraint is 0.1598 113059.094: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 113064.531: recentlyReversed: dir 1 ang 0.000500 revtilt 2.998133 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.012 -0.067 um, 0.9 4.8 % Error in Z Prediction 0.004 Predicted X = -0.430 StdErr = 0.011 nFit = 3 nDrop = 0 Predicted Y = -0.005 StdErr = 0.038 nFit = 3 nDrop = 0 Predicted Z = 4.599 StdErr = 0.001 nFit = 3 nDrop = 0 Need regular tracking - Tracking image shifted 0.005, 0.018 um; position -0.435, -0.023 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0011_0.0.tif Frame alignment results: distance raw = 12.4 smoothed = 12.2 Weighted resid mean = 0.05 max max = 0.07 Mean unweighted max = 0.21 Record image shifted -0.000, 0.000 um Tracking reference shifted by 0.2,-0.0 pixels, -0.000,0.000 um Saved Z = 11 at 5.38 min Tilt = 0.00 Mean = 48 Exp = 1.598 X/Y/Z = -0.435 -0.023 4.599 113087.062: Saved state 11 at 0.0: def 4.499 IS 0.382 0.214 int 0.12073 exp 1.598 113087.062: SetIntensity called with factor 1.0017 113087.062: Changing (ideal) Record exposure by 1.0017 to 1.6006 113087.078: Frame time for constraint is 0.1598 113087.078: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 113090.484: recentlyReversed: dir 1 ang 1.999255 revtilt 4.998888 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.005 -0.018 um, 0.4 1.3 % Predicted X = -0.443 StdErr = 0.011 nFit = 3 nDrop = 0 Predicted Y = -0.015 StdErr = 0.012 nFit = 3 nDrop = 0 Predicted Z = 4.580 StdErr = 0.001 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.004, -0.024 um; position -0.439, 0.008 Measured defocus = 0.01 microns changed by -0.11 to target Tracking image shifted 0.001, -0.000 um; position -0.439, 0.009 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0012_2.0.tif Frame alignment results: distance raw = 14.3 smoothed = 14.3 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.21 Record image shifted -0.000, 0.001 um Tracking reference shifted by 0.2,-1.0 pixels, -0.000,0.001 um Saved Z = 12 at 5.90 min Tilt = 2.00 Mean = 48 Exp = 1.601 X/Y/Z = -0.439 0.008 4.468 113118.250: Saved state 12 at 2.0: def 4.368 IS 0.371 0.243 int 0.12073 exp 1.601 113118.328: Restored state 10 at -22.0 for tiltInd 13 def 5.194 IS 0.524 -0.227 113118.344: SetIntensity called with factor 1.0020 113118.344: Changing (ideal) Record exposure by 1.0020 to 1.6039 113118.359: Frame time for constraint is 0.1598 113118.359: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.004 0.023 um, 0.3 1.6 % Error in Z Prediction -0.112 Predicted X = -0.344 StdErr = 0.021 nFit = 4 nDrop = 0 Predicted Y = -0.560 StdErr = 0.022 nFit = 4 nDrop = 0 Predicted Z = 5.370 StdErr = 0.026 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.014, -0.031 um; position -0.358, -0.530 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0013_-24.0.tif Frame alignment results: distance raw = 7.7 smoothed = 7.2 Weighted resid mean = 0.06 max max = 0.11 Mean unweighted max = 0.16 Record image shifted -0.003, 0.015 um Tracking reference shifted by -0.1,-11.3 pixels, -0.003,0.015 um Saved Z = 13 at 6.34 min Tilt = -23.99 Mean = 49 Exp = 1.604 X/Y/Z = -0.355 -0.545 5.370 113144.547: Saved state 13 at -24.0: def 5.270 IS 0.562 -0.290 int 0.12073 exp 1.604 113144.562: SetIntensity called with factor 1.0024 113144.562: Changing (ideal) Record exposure by 1.0024 to 1.6076 113144.562: Frame time for constraint is 0.1598 113144.578: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.011 0.016 um, 0.8 1.1 % Predicted X = -0.348 StdErr = 0.023 nFit = 4 nDrop = 0 Predicted Y = -0.659 StdErr = 0.015 nFit = 5 nDrop = 0 Predicted Z = 5.448 StdErr = 0.030 nFit = 4 nDrop = 0 Need autofocus Measured defocus = -0.11 microns changed by 0.01 to target 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0014_-26.0.tif Frame alignment results: distance raw = 8.8 smoothed = 8.3 Weighted resid mean = 0.06 max max = 0.09 Mean unweighted max = 0.19 Record image shifted -0.004, -0.037 um Saved Z = 14 at 6.76 min Tilt = -25.99 Mean = 49 Exp = 1.608 X/Y/Z = -0.344 -0.622 5.461 113169.656: Saved state 14 at -26.0: def 5.361 IS 0.588 -0.364 int 0.12073 exp 1.608 113169.734: Restored state 12 at 2.0 for tiltInd 15 def 4.368 IS 0.371 0.243 113169.750: SetIntensity called with factor 1.0020 113169.750: Changing (ideal) Record exposure by 1.0020 to 1.6038 113169.750: Frame time for constraint is 0.1598 113169.765: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 113176.187: recentlyReversed: dir 1 ang 3.999010 revtilt 6.998643 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.004 0.037 um, 0.3 2.7 % Error in Z Prediction 0.013 Predicted X = -0.452 StdErr = 0.010 nFit = 3 nDrop = 0 Predicted Y = 0.030 StdErr = 0.009 nFit = 3 nDrop = 0 Predicted Z = 4.418 StdErr = 0.051 nFit = 3 nDrop = 0 Need regular tracking - Tracking image shifted 0.010, -0.043 um; position -0.462, 0.074 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0015_4.0.tif Frame alignment results: distance raw = 16.2 smoothed = 16.1 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.27 Record image shifted -0.001, 0.001 um Tracking reference shifted by 0.3,-0.8 pixels, -0.001,0.001 um Saved Z = 15 at 7.24 min Tilt = 4.00 Mean = 48 Exp = 1.604 X/Y/Z = -0.462 0.073 4.418 113198.515: Saved state 15 at 4.0: def 4.318 IS 0.359 0.313 int 0.12073 exp 1.604 113198.515: SetIntensity called with factor 1.0023 113198.531: Changing (ideal) Record exposure by 1.0023 to 1.6076 113198.531: Frame time for constraint is 0.1598 113198.547: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 113202.453: recentlyReversed: dir 1 ang 5.999265 revtilt 8.999397 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.010 0.043 um, 0.7 3.1 % Predicted X = -0.479 StdErr = 0.021 nFit = 4 nDrop = 0 Predicted Y = 0.156 StdErr = 0.015 nFit = 4 nDrop = 0 Predicted Z = 4.360 StdErr = 0.062 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.013, 0.035 um; position -0.466, 0.121 Measured defocus = -0.02 microns changed by -0.08 to target Tracking image shifted 0.000, 0.000 um; position -0.467, 0.120 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0016_6.0.tif Frame alignment results: distance raw = 18.3 smoothed = 17.7 Weighted resid mean = 0.07 max max = 0.12 Mean unweighted max = 0.15 Record image shifted -0.000, 0.003 um Tracking reference shifted by 0.0,-1.9 pixels, -0.000,0.003 um Saved Z = 16 at 7.77 min Tilt = 6.00 Mean = 48 Exp = 1.608 X/Y/Z = -0.466 0.118 4.276 113230.234: Saved state 16 at 6.0: def 4.176 IS 0.342 0.355 int 0.12073 exp 1.608 113230.312: Restored state 14 at -26.0 for tiltInd 17 def 5.361 IS 0.588 -0.364 113230.312: SetIntensity called with factor 1.0027 113230.328: Changing (ideal) Record exposure by 1.0027 to 1.6120 113230.328: Frame time for constraint is 0.1598 113230.344: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.013 -0.038 um, 0.9 2.8 % Error in Z Prediction -0.084 Predicted X = -0.339 StdErr = 0.020 nFit = 4 nDrop = 0 Predicted Y = -0.748 StdErr = 0.022 nFit = 6 nDrop = 0 Predicted Z = 5.536 StdErr = 0.021 nFit = 5 nDrop = 0 Need regular tracking - Tracking image shifted 0.019, -0.128 um; position -0.358, -0.620 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0017_-28.0.tif Frame alignment results: distance raw = 8.5 smoothed = 8.4 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.28 Record image shifted -0.003, 0.018 um Tracking reference shifted by -0.1,-13.4 pixels, -0.003,0.018 um Saved Z = 17 at 8.21 min Tilt = -27.99 Mean = 49 Exp = 1.612 X/Y/Z = -0.355 -0.638 5.536 113256.906: Saved state 17 at -28.0: def 5.436 IS 0.606 -0.372 int 0.12073 exp 1.612 113256.922: SetIntensity called with factor 1.0030 113256.922: Changing (ideal) Record exposure by 1.0030 to 1.6168 113256.937: Frame time for constraint is 0.1598 113256.937: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.016 0.111 um, 1.2 8.0 % Predicted X = -0.361 StdErr = 0.026 nFit = 4 nDrop = 0 Predicted Y = -0.676 StdErr = 0.041 nFit = 5 nDrop = 1 Predicted Z = 5.616 StdErr = 0.023 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.008, -0.059 um; position -0.353, -0.617 Measured defocus = -0.10 microns changed by 0.00 to target Tracking image shifted 0.001, -0.002 um; position -0.354, -0.615 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0018_-30.0.tif Frame alignment results: distance raw = 11.2 smoothed = 10.7 Weighted resid mean = 0.08 max max = 0.12 Mean unweighted max = 0.19 Record image shifted -0.002, 0.011 um Tracking reference shifted by -0.4,-8.3 pixels, -0.002,0.011 um Saved Z = 18 at 8.69 min Tilt = -29.99 Mean = 49 Exp = 1.617 X/Y/Z = -0.352 -0.626 5.619 113285.672: Saved state 18 at -30.0: def 5.519 IS 0.598 -0.363 int 0.12073 exp 1.617 113285.750: Restored state 16 at 6.0 for tiltInd 19 def 4.176 IS 0.342 0.355 113285.765: SetIntensity called with factor 1.0027 113285.765: Changing (ideal) Record exposure by 1.0027 to 1.6119 113285.781: Frame time for constraint is 0.1598 113285.781: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 113292.672: recentlyReversed: dir 1 ang 7.999020 revtilt 10.990154 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.008 0.050 um, 0.6 3.6 % Error in Z Prediction 0.004 Predicted X = -0.480 StdErr = 0.024 nFit = 4 nDrop = 0 Predicted Y = 0.195 StdErr = 0.023 nFit = 5 nDrop = 0 Predicted Z = 4.222 StdErr = 0.054 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.006, 0.031 um; position -0.486, 0.163 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0019_8.0.tif Frame alignment results: distance raw = 16.4 smoothed = 16.0 Weighted resid mean = 0.07 max max = 0.13 Mean unweighted max = 0.30 Record image shifted -0.001, 0.002 um Tracking reference shifted by 0.4,-1.8 pixels, -0.001,0.002 um Saved Z = 19 at 9.18 min Tilt = 8.00 Mean = 48 Exp = 1.612 X/Y/Z = -0.485 0.161 4.222 113314.922: Saved state 19 at 8.0: def 4.122 IS 0.337 0.404 int 0.12073 exp 1.612 113314.937: SetIntensity called with factor 1.0030 113314.937: Changing (ideal) Record exposure by 1.0030 to 1.6167 113314.953: Frame time for constraint is 0.1598 113314.953: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 113318.344: recentlyReversed: dir 1 ang 9.999275 revtilt 12.998408 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.005 -0.034 um, 0.3 2.4 % Predicted X = -0.494 StdErr = 0.011 nFit = 3 nDrop = 0 Predicted Y = 0.229 StdErr = 0.023 nFit = 6 nDrop = 0 Predicted Z = 4.148 StdErr = 0.060 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.010, 0.056 um; position -0.484, 0.173 Measured defocus = -0.07 microns changed by -0.03 to target Tracking image shifted 0.001, -0.000 um; position -0.485, 0.174 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0020_10.0.tif Frame alignment results: distance raw = 19.9 smoothed = 18.7 Weighted resid mean = 0.07 max max = 0.11 Mean unweighted max = 0.27 Record image shifted -0.001, 0.005 um Tracking reference shifted by -0.0,-3.7 pixels, -0.001,0.005 um Saved Z = 20 at 9.70 min Tilt = 10.00 Mean = 48 Exp = 1.617 X/Y/Z = -0.484 0.169 4.119 113346.187: Saved state 20 at 10.0: def 4.019 IS 0.333 0.410 int 0.12073 exp 1.617 113346.265: Restored state 18 at -30.0 for tiltInd 21 def 5.519 IS 0.598 -0.363 113346.265: SetIntensity called with factor 1.0034 113346.281: Changing (ideal) Record exposure by 1.0034 to 1.6223 113346.281: Frame time for constraint is 0.1598 113346.297: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.010 -0.060 um, 0.7 4.4 % Error in Z Prediction -0.029 Predicted X = -0.363 StdErr = 0.025 nFit = 4 nDrop = 0 Predicted Y = -0.585 StdErr = 0.026 nFit = 5 nDrop = 1 Predicted Z = 5.705 StdErr = 0.006 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.001, 0.060 um; position -0.364, -0.645 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0021_-32.0.tif Frame alignment results: distance raw = 11.7 smoothed = 11.3 Weighted resid mean = 0.07 max max = 0.13 Mean unweighted max = 0.23 Record image shifted -0.001, 0.010 um Tracking reference shifted by -0.6,-7.7 pixels, -0.001,0.010 um Saved Z = 21 at 10.16 min Tilt = -31.99 Mean = 49 Exp = 1.622 X/Y/Z = -0.363 -0.656 5.705 113373.500: Saved state 21 at -32.0: def 5.605 IS 0.621 -0.384 int 0.12073 exp 1.622 113373.515: SetIntensity called with factor 1.0037 113373.515: Changing (ideal) Record exposure by 1.0037 to 1.6284 113373.531: Frame time for constraint is 0.1598 113373.531: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.000 -0.070 um, 0.0 5.1 % Predicted X = -0.365 StdErr = 0.010 nFit = 3 nDrop = 0 Predicted Y = -0.615 StdErr = 0.040 nFit = 6 nDrop = 0 Predicted Z = 5.787 StdErr = 0.007 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.013, 0.085 um; position -0.352, -0.701 Measured defocus = 0.01 microns changed by -0.11 to target Tracking image shifted 0.002, -0.001 um; position -0.353, -0.699 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0022_-34.0.tif Frame alignment results: distance raw = 12.7 smoothed = 12.0 Weighted resid mean = 0.08 max max = 0.12 Mean unweighted max = 0.20 Record image shifted -0.002, 0.013 um Tracking reference shifted by -0.3,-9.4 pixels, -0.002,0.013 um Saved Z = 22 at 10.64 min Tilt = -33.99 Mean = 49 Exp = 1.628 X/Y/Z = -0.352 -0.712 5.681 113402.687: Saved state 22 at -34.0: def 5.581 IS 0.638 -0.440 int 0.12073 exp 1.628 113402.781: Restored state 20 at 10.0 for tiltInd 23 def 4.019 IS 0.333 0.410 113402.781: SetIntensity called with factor 1.0034 113402.797: Changing (ideal) Record exposure by 1.0034 to 1.6221 113402.797: Frame time for constraint is 0.1598 113402.812: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 113410.094: recentlyReversed: dir 1 ang 11.999530 revtilt 15.000662 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.013 -0.097 um, 1.0 7.0 % Error in Z Prediction -0.107 Predicted X = -0.497 StdErr = 0.014 nFit = 3 nDrop = 0 Predicted Y = 0.170 StdErr = 0.013 nFit = 5 nDrop = 1 Predicted Z = 4.033 StdErr = 0.019 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.004, -0.020 um; position -0.500, 0.190 Measured defocus = -0.10 microns changed by -0.00 to target Tracking image shifted 0.000, -0.000 um; position -0.500, 0.190 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0023_12.0.tif Frame alignment results: distance raw = 20.5 smoothed = 18.2 Weighted resid mean = 0.07 max max = 0.12 Mean unweighted max = 0.33 Record image shifted -0.002, 0.004 um Tracking reference shifted by 0.6,-3.4 pixels, -0.002,0.004 um Saved Z = 23 at 11.23 min Tilt = 12.00 Mean = 48 Exp = 1.622 X/Y/Z = -0.499 0.186 4.030 113437.797: Saved state 23 at 12.0: def 3.930 IS 0.337 0.433 int 0.12073 exp 1.622 113437.797: SetIntensity called with factor 1.0037 113437.812: Changing (ideal) Record exposure by 1.0037 to 1.6282 113437.812: Frame time for constraint is 0.1598 113437.828: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 113441.172: recentlyReversed: dir 1 ang 13.999285 revtilt 16.993918 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.002 0.016 um, 0.2 1.1 % Error in Z Prediction -0.003 Predicted X = -0.503 StdErr = 0.011 nFit = 3 nDrop = 0 Predicted Y = 0.179 StdErr = 0.017 nFit = 5 nDrop = 0 Predicted Z = 3.942 StdErr = 0.015 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted -0.005, -0.038 um; position -0.498, 0.217 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0024_14.0.tif Frame alignment results: distance raw = 20.2 smoothed = 16.9 Weighted resid mean = 0.08 max max = 0.12 Mean unweighted max = 0.27 Record image shifted -0.000, 0.003 um Tracking reference shifted by -0.2,-2.4 pixels, -0.000,0.003 um Saved Z = 24 at 11.66 min Tilt = 14.00 Mean = 47 Exp = 1.628 X/Y/Z = -0.498 0.213 3.942 113463.406: Saved state 24 at 14.0: def 3.842 IS 0.323 0.457 int 0.12073 exp 1.628 113463.484: Restored state 22 at -34.0 for tiltInd 25 def 5.581 IS 0.638 -0.440 113463.500: SetIntensity called with factor 1.0041 113463.500: Changing (ideal) Record exposure by 1.0041 to 1.6350 113463.515: Frame time for constraint is 0.1598 113463.515: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.005 0.034 um, 0.4 2.5 % Predicted X = -0.355 StdErr = 0.017 nFit = 3 nDrop = 0 Predicted Y = -0.771 StdErr = 0.033 nFit = 5 nDrop = 0 Predicted Z = 5.778 StdErr = 0.057 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted -0.000, 0.013 um; position -0.354, -0.785 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0025_-36.0.tif Frame alignment results: distance raw = 15.0 smoothed = 14.6 Weighted resid mean = 0.07 max max = 0.12 Mean unweighted max = 0.20 Record image shifted -0.000, 0.011 um Tracking reference shifted by -1.1,-8.0 pixels, -0.000,0.011 um Saved Z = 25 at 12.12 min Tilt = -35.99 Mean = 49 Exp = 1.635 X/Y/Z = -0.354 -0.796 5.778 113491.359: Saved state 25 at -36.0: def 5.678 IS 0.680 -0.512 int 0.12073 exp 1.635 113491.359: SetIntensity called with factor 1.0045 113491.375: Changing (ideal) Record exposure by 1.0045 to 1.6424 113491.375: Frame time for constraint is 0.1598 113491.390: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.001 -0.024 um, 0.1 1.8 % Predicted X = -0.343 StdErr = 0.024 nFit = 4 nDrop = 0 Predicted Y = -0.915 StdErr = 0.008 nFit = 5 nDrop = 1 Predicted Z = 5.844 StdErr = 0.063 nFit = 4 nDrop = 0 Need autofocus Measured defocus = -0.06 microns changed by -0.04 to target 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0026_-38.0.tif Frame alignment results: distance raw = 15.6 smoothed = 15.2 Weighted resid mean = 0.06 max max = 0.10 Mean unweighted max = 0.32 Record image shifted -0.000, -0.054 um Saved Z = 26 at 12.54 min Tilt = -37.99 Mean = 49 Exp = 1.642 X/Y/Z = -0.343 -0.861 5.800 113516.687: Saved state 26 at -38.0: def 5.700 IS 0.702 -0.576 int 0.12073 exp 1.642 113516.765: Restored state 24 at 14.0 for tiltInd 27 def 3.842 IS 0.323 0.457 113516.765: SetIntensity called with factor 1.0041 113516.781: Changing (ideal) Record exposure by 1.0041 to 1.6348 113516.781: Frame time for constraint is 0.1598 113516.797: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 113524.047: recentlyReversed: dir 1 ang 15.999040 revtilt 18.998172 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.000 0.054 um, 0.0 3.9 % Error in Z Prediction -0.043 Predicted X = -0.507 StdErr = 0.011 nFit = 3 nDrop = 0 Predicted Y = 0.223 StdErr = 0.026 nFit = 5 nDrop = 0 Predicted Z = 3.855 StdErr = 0.017 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.015, -0.071 um; position -0.523, 0.294 Measured defocus = -0.06 microns changed by -0.04 to target Tracking image shifted -0.000, -0.000 um; position -0.523, 0.295 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0027_16.0.tif Frame alignment results: distance raw = 20.5 smoothed = 16.3 Weighted resid mean = 0.10 max max = 0.16 Mean unweighted max = 0.26 Record image shifted -0.002, 0.007 um Tracking reference shifted by 0.2,-5.1 pixels, -0.002,0.007 um Saved Z = 27 at 13.13 min Tilt = 16.00 Mean = 47 Exp = 1.635 X/Y/Z = -0.521 0.288 3.813 113551.781: Saved state 27 at 16.0: def 3.713 IS 0.308 0.535 int 0.12073 exp 1.635 113551.797: SetIntensity called with factor 1.0045 113551.797: Changing (ideal) Record exposure by 1.0045 to 1.6421 113551.812: Frame time for constraint is 0.1598 113551.828: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 113555.187: recentlyReversed: dir 1 ang 17.999795 revtilt 20.993928 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.014 0.065 um, 1.0 4.7 % Error in Z Prediction -0.042 Predicted X = -0.528 StdErr = 0.018 nFit = 3 nDrop = 0 Predicted Y = 0.365 StdErr = 0.021 nFit = 5 nDrop = 0 Predicted Z = 3.736 StdErr = 0.032 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.005, 0.006 um; position -0.523, 0.359 Measured defocus = -0.09 microns changed by -0.01 to target Tracking image shifted 0.000, 0.001 um; position -0.523, 0.358 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0028_18.0.tif Frame alignment results: distance raw = 20.1 smoothed = 15.3 Weighted resid mean = 0.09 max max = 0.14 Mean unweighted max = 0.38 Record image shifted -0.001, 0.006 um Tracking reference shifted by -0.4,-4.4 pixels, -0.001,0.006 um Saved Z = 28 at 13.65 min Tilt = 18.00 Mean = 47 Exp = 1.642 X/Y/Z = -0.522 0.352 3.723 113582.859: Saved state 28 at 18.0: def 3.623 IS 0.279 0.593 int 0.12073 exp 1.642 113582.953: Restored state 26 at -38.0 for tiltInd 29 def 5.700 IS 0.702 -0.576 113582.953: SetIntensity called with factor 1.0049 113582.969: Changing (ideal) Record exposure by 1.0049 to 1.6504 113582.969: Frame time for constraint is 0.1598 113582.984: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.006 -0.013 um, 0.4 0.9 % Error in Z Prediction -0.014 Predicted X = -0.341 StdErr = 0.010 nFit = 3 nDrop = 0 Predicted Y = -0.963 StdErr = 0.025 nFit = 5 nDrop = 0 Predicted Z = 5.836 StdErr = 0.037 nFit = 3 nDrop = 0 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0029_-40.0.tif Frame alignment results: distance raw = 15.5 smoothed = 13.2 Weighted resid mean = 0.07 max max = 0.12 Mean unweighted max = 0.30 Record image shifted 0.013, -0.060 um Saved Z = 29 at 14.01 min Tilt = -39.99 Mean = 48 Exp = 1.650 X/Y/Z = -0.354 -0.902 5.836 113604.390: Saved state 29 at -40.0: def 5.736 IS 0.730 -0.606 int 0.12073 exp 1.650 113604.406: SetIntensity called with factor 1.0053 113604.406: Changing (ideal) Record exposure by 1.0053 to 1.6591 113604.422: Frame time for constraint is 0.1598 113604.422: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.013 0.060 um, 0.9 4.4 % Predicted X = -0.360 StdErr = 0.023 nFit = 4 nDrop = 0 Predicted Y = -0.954 StdErr = 0.029 nFit = 5 nDrop = 0 Predicted Z = 5.881 StdErr = 0.043 nFit = 3 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.006, -0.078 um; position -0.354, -0.875 Measured defocus = -0.19 microns changed by 0.09 to target Tracking image shifted 0.001, -0.003 um; position -0.355, -0.872 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0030_-42.0.tif Frame alignment results: distance raw = 18.1 smoothed = 15.1 Weighted resid mean = 0.11 max max = 0.17 Mean unweighted max = 0.32 Record image shifted -0.005, 0.044 um Tracking reference shifted by -2.1,-32.6 pixels, -0.005,0.044 um Saved Z = 30 at 14.49 min Tilt = -41.99 Mean = 48 Exp = 1.659 X/Y/Z = -0.350 -0.916 5.973 113633.578: Saved state 30 at -42.0: def 5.873 IS 0.733 -0.621 int 0.12073 exp 1.659 113633.672: Restored state 28 at 18.0 for tiltInd 31 def 3.623 IS 0.279 0.593 113633.672: SetIntensity called with factor 1.0049 113633.687: Changing (ideal) Record exposure by 1.0049 to 1.6501 113633.703: Frame time for constraint is 0.1598 113633.703: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 113641.219: recentlyReversed: dir 1 ang 19.999050 revtilt 22.998682 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.010 0.038 um, 0.7 2.7 % Error in Z Prediction 0.092 Predicted X = -0.536 StdErr = 0.030 nFit = 4 nDrop = 0 Predicted Y = 0.452 StdErr = 0.022 nFit = 5 nDrop = 0 Predicted Z = 3.618 StdErr = 0.012 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.014, 0.001 um; position -0.550, 0.451 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0031_20.0.tif Frame alignment results: distance raw = 18.2 smoothed = 13.8 Weighted resid mean = 0.08 max max = 0.13 Mean unweighted max = 0.30 Record image shifted -0.001, 0.006 um Tracking reference shifted by 0.1,-4.5 pixels, -0.001,0.006 um Saved Z = 31 at 14.99 min Tilt = 20.00 Mean = 47 Exp = 1.650 X/Y/Z = -0.549 0.445 3.618 113663.531: Saved state 31 at 20.0: def 3.518 IS 0.257 0.689 int 0.12073 exp 1.650 113663.547: SetIntensity called with factor 1.0053 113663.547: Changing (ideal) Record exposure by 1.0053 to 1.6587 113663.562: Frame time for constraint is 0.1598 113663.578: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 113666.890: recentlyReversed: dir 1 ang 22.000305 revtilt 24.982939 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.013 -0.007 um, 0.9 0.5 % Predicted X = -0.559 StdErr = 0.019 nFit = 3 nDrop = 0 Predicted Y = 0.554 StdErr = 0.022 nFit = 5 nDrop = 0 Predicted Z = 3.517 StdErr = 0.014 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.012, 0.080 um; position -0.547, 0.474 Measured defocus = -0.13 microns changed by 0.03 to target Tracking image shifted 0.001, -0.001 um; position -0.548, 0.475 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0032_22.0.tif Frame alignment results: distance raw = 19.6 smoothed = 15.1 Weighted resid mean = 0.10 max max = 0.16 Mean unweighted max = 0.32 Record image shifted -0.002, 0.009 um Tracking reference shifted by -0.0,-6.5 pixels, -0.002,0.009 um Saved Z = 32 at 15.52 min Tilt = 22.00 Mean = 47 Exp = 1.659 X/Y/Z = -0.547 0.467 3.550 113695.328: Saved state 32 at 22.0: def 3.450 IS 0.245 0.707 int 0.12073 exp 1.659 113695.406: Restored state 30 at -42.0 for tiltInd 33 def 5.873 IS 0.733 -0.621 113695.422: SetIntensity called with factor 1.0057 113695.422: Changing (ideal) Record exposure by 1.0057 to 1.6686 113695.437: Frame time for constraint is 0.1598 113695.453: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.012 -0.087 um, 0.9 6.3 % Error in Z Prediction 0.033 Predicted X = -0.359 StdErr = 0.024 nFit = 4 nDrop = 0 Predicted Y = -0.910 StdErr = 0.005 nFit = 5 nDrop = 0 Predicted Z = 6.038 StdErr = 0.034 nFit = 3 nDrop = 0 Need regular tracking - Tracking image shifted 0.008, -0.003 um; position -0.367, -0.907 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0033_-44.0.tif Frame alignment results: distance raw = 18.2 smoothed = 14.5 Weighted resid mean = 0.08 max max = 0.15 Mean unweighted max = 0.39 Record image shifted -0.001, 0.014 um Tracking reference shifted by -1.3,-10.6 pixels, -0.001,0.014 um Saved Z = 33 at 16.00 min Tilt = -43.99 Mean = 48 Exp = 1.669 X/Y/Z = -0.366 -0.922 6.038 113724.078: Saved state 33 at -44.0: def 5.938 IS 0.750 -0.617 int 0.12073 exp 1.669 113724.094: SetIntensity called with factor 1.0062 113724.109: Changing (ideal) Record exposure by 1.0062 to 1.6789 113724.109: Frame time for constraint is 0.1598 113724.125: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.007 -0.012 um, 0.5 0.9 % Predicted X = -0.376 StdErr = 0.023 nFit = 4 nDrop = 0 Predicted Y = -0.897 StdErr = 0.007 nFit = 6 nDrop = 0 Predicted Z = 6.111 StdErr = 0.040 nFit = 3 nDrop = 0 Need autofocus Measured defocus = -0.10 microns changed by 0.00 to target 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0034_-46.0.tif Frame alignment results: distance raw = 17.0 smoothed = 13.1 Weighted resid mean = 0.10 max max = 0.16 Mean unweighted max = 0.36 Record image shifted -0.013, 0.077 um Saved Z = 34 at 16.43 min Tilt = -45.98 Mean = 48 Exp = 1.679 X/Y/Z = -0.363 -0.974 6.113 113749.609: Saved state 34 at -46.0: def 6.013 IS 0.771 -0.665 int 0.12073 exp 1.679 113749.703: Restored state 32 at 22.0 for tiltInd 35 def 3.450 IS 0.245 0.707 113749.703: SetIntensity called with factor 1.0057 113749.719: Changing (ideal) Record exposure by 1.0057 to 1.6682 113749.734: Frame time for constraint is 0.1598 113749.734: Will be constrained to 1.5979 (frame 0.1598, constraint 0.1598) 113757.531: recentlyReversed: dir 1 ang 23.999060 revtilt 26.998192 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.013 -0.077 um, 0.9 5.5 % Error in Z Prediction 0.003 Predicted X = -0.564 StdErr = 0.022 nFit = 3 nDrop = 0 Predicted Y = 0.549 StdErr = 0.046 nFit = 6 nDrop = 0 Predicted Z = 3.444 StdErr = 0.024 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.002, 0.061 um; position -0.565, 0.488 10 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0035_24.0.tif Frame alignment results: distance raw = 18.5 smoothed = 15.7 Weighted resid mean = 0.10 max max = 0.17 Mean unweighted max = 0.31 Record image shifted -0.001, 0.007 um Tracking reference shifted by -0.1,-5.4 pixels, -0.001,0.007 um Saved Z = 35 at 16.93 min Tilt = 24.00 Mean = 46 Exp = 1.668 X/Y/Z = -0.564 0.480 3.444 113779.781: Saved state 35 at 24.0: def 3.344 IS 0.253 0.729 int 0.12073 exp 1.668 113779.797: SetIntensity called with factor 1.0061 113779.812: Changing (ideal) Record exposure by 1.0061 to 1.6784 113779.828: Frame time for constraint is 0.1598 113779.828: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 113783.047: recentlyReversed: dir 1 ang 25.999315 revtilt 28.998447 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.001 -0.069 um, 0.1 5.0 % Predicted X = -0.565 StdErr = 0.033 nFit = 4 nDrop = 0 Predicted Y = 0.470 StdErr = 0.038 nFit = 5 nDrop = 0 Predicted Z = 3.348 StdErr = 0.027 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.003, -0.026 um; position -0.562, 0.496 Measured defocus = -0.11 microns changed by 0.01 to target Tracking image shifted -0.000, -0.001 um; position -0.562, 0.497 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0036_26.0.tif Frame alignment results: distance raw = 21.6 smoothed = 17.7 Weighted resid mean = 0.09 max max = 0.15 Mean unweighted max = 0.42 Record image shifted -0.001, 0.011 um Tracking reference shifted by -0.6,-7.9 pixels, -0.001,0.011 um Saved Z = 36 at 17.45 min Tilt = 26.00 Mean = 46 Exp = 1.678 X/Y/Z = -0.560 0.487 3.353 113811.140: Saved state 36 at 26.0: def 3.253 IS 0.247 0.732 int 0.12073 exp 1.678 113811.219: Restored state 34 at -46.0 for tiltInd 37 def 6.013 IS 0.771 -0.665 113811.234: SetIntensity called with factor 1.0066 113811.250: Changing (ideal) Record exposure by 1.0066 to 1.6901 113811.250: Frame time for constraint is 0.1598 113811.265: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.005 0.016 um, 0.3 1.2 % Error in Z Prediction 0.005 Predicted X = -0.369 StdErr = 0.026 nFit = 4 nDrop = 0 Predicted Y = -0.968 StdErr = 0.039 nFit = 6 nDrop = 0 Predicted Z = 6.186 StdErr = 0.022 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.017, 0.039 um; position -0.386, -1.008 Measured defocus = -0.03 microns changed by -0.07 to target Tracking image shifted 0.001, -0.001 um; position -0.387, -1.007 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0037_-48.0.tif Frame alignment results: distance raw = 19.9 smoothed = 15.8 Weighted resid mean = 0.10 max max = 0.15 Mean unweighted max = 0.29 Record image shifted -0.005, 0.031 um Tracking reference shifted by -0.4,-23.5 pixels, -0.005,0.031 um Saved Z = 37 at 18.03 min Tilt = -47.99 Mean = 48 Exp = 1.690 X/Y/Z = -0.382 -1.038 6.120 113845.953: Saved state 37 at -48.0: def 6.020 IS 0.818 -0.711 int 0.12073 exp 1.690 113845.953: SetIntensity called with factor 1.0071 113845.969: Changing (ideal) Record exposure by 1.0071 to 1.7021 113845.984: Frame time for constraint is 0.1598 113846.000: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.012 -0.069 um, 0.9 5.0 % Error in Z Prediction -0.066 Predicted X = -0.392 StdErr = 0.027 nFit = 4 nDrop = 0 Predicted Y = -1.123 StdErr = 0.021 nFit = 5 nDrop = 1 Predicted Z = 6.218 StdErr = 0.050 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.019, -0.022 um; position -0.373, -1.101 Measured defocus = -0.03 microns changed by -0.07 to target Tracking image shifted 0.001, -0.002 um; position -0.374, -1.099 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0038_-50.0.tif Frame alignment results: distance raw = 18.9 smoothed = 16.1 Weighted resid mean = 0.10 max max = 0.18 Mean unweighted max = 0.44 Record image shifted -0.003, 0.016 um Tracking reference shifted by -0.2,-11.9 pixels, -0.003,0.016 um Saved Z = 38 at 18.52 min Tilt = -49.99 Mean = 48 Exp = 1.702 X/Y/Z = -0.371 -1.115 6.152 113875.125: Saved state 38 at -50.0: def 6.052 IS 0.846 -0.785 int 0.12073 exp 1.702 113875.219: Restored state 36 at 26.0 for tiltInd 39 def 3.253 IS 0.247 0.732 113875.219: SetIntensity called with factor 1.0066 113875.234: Changing (ideal) Record exposure by 1.0066 to 1.6895 113875.250: Frame time for constraint is 0.1598 113875.265: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 113883.390: recentlyReversed: dir 1 ang 27.999070 revtilt 30.998202 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.021 0.008 um, 1.5 0.6 % Error in Z Prediction -0.066 Predicted X = -0.571 StdErr = 0.016 nFit = 3 nDrop = 0 Predicted Y = 0.447 StdErr = 0.036 nFit = 5 nDrop = 0 Predicted Z = 3.267 StdErr = 0.011 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.017, -0.095 um; position -0.588, 0.542 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0039_28.0.tif Frame alignment results: distance raw = 22.8 smoothed = 20.0 Weighted resid mean = 0.11 max max = 0.17 Mean unweighted max = 0.36 Record image shifted -0.002, 0.008 um Tracking reference shifted by 0.6,-6.2 pixels, -0.002,0.008 um Saved Z = 39 at 19.03 min Tilt = 28.00 Mean = 45 Exp = 1.689 X/Y/Z = -0.586 0.534 3.267 113906.172: Saved state 39 at 28.0: def 3.167 IS 0.246 0.787 int 0.12073 exp 1.689 113906.187: SetIntensity called with factor 1.0071 113906.203: Changing (ideal) Record exposure by 1.0071 to 1.7015 113906.219: Frame time for constraint is 0.1598 113906.234: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 113909.422: recentlyReversed: dir 1 ang 29.999325 revtilt 32.981459 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.015 0.087 um, 1.1 6.3 % Predicted X = -0.603 StdErr = 0.034 nFit = 4 nDrop = 0 Predicted Y = 0.563 StdErr = 0.027 nFit = 5 nDrop = 1 Predicted Z = 3.175 StdErr = 0.013 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.020, 0.005 um; position -0.583, 0.558 Measured defocus = 0.00 microns changed by -0.10 to target Tracking image shifted -0.001, 0.003 um; position -0.582, 0.555 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0040_30.0.tif Frame alignment results: distance raw = 23.0 smoothed = 19.6 Weighted resid mean = 0.11 max max = 0.17 Mean unweighted max = 0.45 Record image shifted -0.000, 0.013 um Tracking reference shifted by -1.5,-9.2 pixels, -0.000,0.013 um Saved Z = 40 at 19.56 min Tilt = 30.00 Mean = 45 Exp = 1.701 X/Y/Z = -0.582 0.543 3.076 113937.515: Saved state 40 at 30.0: def 2.976 IS 0.239 0.793 int 0.12073 exp 1.701 113937.609: Restored state 38 at -50.0 for tiltInd 41 def 6.052 IS 0.846 -0.785 113937.609: SetIntensity called with factor 1.0077 113937.625: Changing (ideal) Record exposure by 1.0077 to 1.7152 113937.640: Frame time for constraint is 0.1598 113937.656: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.021 -0.020 um, 1.5 1.5 % Error in Z Prediction -0.099 Predicted X = -0.370 StdErr = 0.034 nFit = 4 nDrop = 0 Predicted Y = -1.217 StdErr = 0.015 nFit = 6 nDrop = 0 Predicted Z = 6.244 StdErr = 0.053 nFit = 5 nDrop = 0 Need regular tracking - Tracking image shifted 0.025, -0.087 um; position -0.395, -1.130 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0041_-52.0.tif Frame alignment results: distance raw = 19.6 smoothed = 18.1 Weighted resid mean = 0.12 max max = 0.20 Mean unweighted max = 0.33 Record image shifted -0.002, 0.018 um Tracking reference shifted by -0.6,-13.1 pixels, -0.002,0.018 um Saved Z = 41 at 20.05 min Tilt = -51.98 Mean = 47 Exp = 1.715 X/Y/Z = -0.393 -1.148 6.244 113966.953: Saved state 41 at -52.0: def 6.144 IS 0.880 -0.803 int 0.12073 exp 1.715 113966.969: SetIntensity called with factor 1.0082 113966.984: Changing (ideal) Record exposure by 1.0082 to 1.7293 113967.000: Frame time for constraint is 0.1598 113967.015: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.022 0.070 um, 1.6 5.0 % Predicted X = -0.393 StdErr = 0.024 nFit = 3 nDrop = 0 Predicted Y = -1.238 StdErr = 0.041 nFit = 6 nDrop = 0 Predicted Z = 6.303 StdErr = 0.058 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.002, -0.075 um; position -0.391, -1.164 Measured defocus = -0.06 microns changed by -0.04 to target Tracking image shifted 0.001, -0.002 um; position -0.392, -1.162 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0042_-54.0.tif Frame alignment results: distance raw = 18.6 smoothed = 17.2 Weighted resid mean = 0.10 max max = 0.14 Mean unweighted max = 0.36 Record image shifted -0.003, 0.020 um Tracking reference shifted by -0.3,-14.9 pixels, -0.003,0.020 um Saved Z = 42 at 20.54 min Tilt = -53.98 Mean = 47 Exp = 1.729 X/Y/Z = -0.389 -1.182 6.271 113996.297: Saved state 42 at -54.0: def 6.171 IS 0.893 -0.835 int 0.12073 exp 1.729 113996.390: Restored state 40 at 30.0 for tiltInd 43 def 2.976 IS 0.239 0.793 113996.406: SetIntensity called with factor 1.0076 113996.406: Changing (ideal) Record exposure by 1.0076 to 1.7144 113996.422: Frame time for constraint is 0.1598 113996.437: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 114004.828: recentlyReversed: dir 1 ang 31.999080 revtilt 34.993713 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.004 0.057 um, 0.3 4.1 % Error in Z Prediction -0.033 Predicted X = -0.597 StdErr = 0.022 nFit = 3 nDrop = 0 Predicted Y = 0.579 StdErr = 0.030 nFit = 5 nDrop = 0 Predicted Z = 2.998 StdErr = 0.054 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.013, -0.019 um; position -0.610, 0.598 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0043_32.0.tif Frame alignment results: distance raw = 26.1 smoothed = 24.4 Weighted resid mean = 0.13 max max = 0.20 Mean unweighted max = 0.32 Record image shifted -0.003, 0.007 um Tracking reference shifted by 1.0,-5.7 pixels, -0.003,0.007 um Saved Z = 43 at 21.06 min Tilt = 32.00 Mean = 44 Exp = 1.714 X/Y/Z = -0.608 0.591 2.998 114027.578: Saved state 43 at 32.0: def 2.898 IS 0.238 0.849 int 0.12073 exp 1.714 114027.594: SetIntensity called with factor 1.0082 114027.609: Changing (ideal) Record exposure by 1.0082 to 1.7284 114027.625: Frame time for constraint is 0.1598 114027.640: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 114030.890: recentlyReversed: dir 1 ang 33.999335 revtilt 36.987469 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.010 0.011 um, 0.7 0.8 % Predicted X = -0.613 StdErr = 0.022 nFit = 3 nDrop = 0 Predicted Y = 0.632 StdErr = 0.022 nFit = 6 nDrop = 0 Predicted Z = 2.892 StdErr = 0.060 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.009, 0.025 um; position -0.605, 0.608 Measured defocus = -0.11 microns changed by 0.01 to target Tracking image shifted 0.000, -0.000 um; position -0.605, 0.608 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0044_34.0.tif Frame alignment results: distance raw = 24.8 smoothed = 21.4 Weighted resid mean = 0.13 max max = 0.21 Mean unweighted max = 0.51 Record image shifted -0.001, 0.018 um Tracking reference shifted by -1.6,-13.0 pixels, -0.001,0.018 um Saved Z = 44 at 21.58 min Tilt = 34.00 Mean = 44 Exp = 1.728 X/Y/Z = -0.604 0.590 2.897 114058.875: Saved state 44 at 34.0: def 2.797 IS 0.235 0.847 int 0.12073 exp 1.728 114058.969: Restored state 42 at -54.0 for tiltInd 45 def 6.171 IS 0.893 -0.835 114058.984: SetIntensity called with factor 1.0088 114059.000: Changing (ideal) Record exposure by 1.0088 to 1.7446 114059.015: Frame time for constraint is 0.1598 114059.031: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.010 -0.042 um, 0.7 3.0 % Error in Z Prediction 0.005 Predicted X = -0.403 StdErr = 0.038 nFit = 4 nDrop = 0 Predicted Y = -1.217 StdErr = 0.025 nFit = 6 nDrop = 0 Predicted Z = 6.312 StdErr = 0.035 nFit = 5 nDrop = 0 Need regular tracking - Tracking image shifted 0.017, -0.078 um; position -0.420, -1.139 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0045_-56.0.tif Frame alignment results: distance raw = 21.3 smoothed = 20.8 Weighted resid mean = 0.11 max max = 0.17 Mean unweighted max = 0.29 Record image shifted -0.003, 0.019 um Tracking reference shifted by -0.6,-14.1 pixels, -0.003,0.019 um Saved Z = 45 at 22.08 min Tilt = -55.98 Mean = 47 Exp = 1.745 X/Y/Z = -0.417 -1.158 6.312 114089.125: Saved state 45 at -56.0: def 6.212 IS 0.905 -0.798 int 0.12073 exp 1.745 114089.140: SetIntensity called with factor 1.0095 114089.156: Changing (ideal) Record exposure by 1.0095 to 1.7611 114089.172: Frame time for constraint is 0.1598 114089.187: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.014 0.059 um, 1.0 4.3 % Predicted X = -0.424 StdErr = 0.024 nFit = 3 nDrop = 0 Predicted Y = -1.120 StdErr = 0.022 nFit = 5 nDrop = 0 Predicted Z = 6.359 StdErr = 0.038 nFit = 5 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.007, 0.040 um; position -0.417, -1.160 Measured defocus = -0.17 microns changed by 0.07 to target Tracking image shifted 0.000, -0.001 um; position -0.418, -1.158 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0046_-58.0.tif Frame alignment results: distance raw = 20.8 smoothed = 19.2 Weighted resid mean = 0.11 max max = 0.17 Mean unweighted max = 0.44 Record image shifted -0.003, 0.022 um Tracking reference shifted by -0.6,-16.6 pixels, -0.003,0.022 um Saved Z = 46 at 22.58 min Tilt = -57.99 Mean = 47 Exp = 1.761 X/Y/Z = -0.414 -1.180 6.434 114118.609: Saved state 46 at -58.0: def 6.334 IS 0.914 -0.820 int 0.12073 exp 1.761 114118.719: Restored state 44 at 34.0 for tiltInd 47 def 2.797 IS 0.235 0.847 114118.734: SetIntensity called with factor 1.0088 114118.750: Changing (ideal) Record exposure by 1.0088 to 1.7436 114118.750: Frame time for constraint is 0.1598 114118.781: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 114127.500: recentlyReversed: dir 1 ang 35.999590 revtilt 38.998722 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 0.010 -0.061 um, 0.7 4.4 % Error in Z Prediction 0.076 Predicted X = -0.620 StdErr = 0.022 nFit = 3 nDrop = 0 Predicted Y = 0.600 StdErr = 0.033 nFit = 5 nDrop = 0 Predicted Z = 2.772 StdErr = 0.041 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.017, -0.078 um; position -0.637, 0.679 Measured defocus = -0.10 microns changed by -0.00 to target Tracking image shifted -0.001, -0.001 um; position -0.636, 0.680 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0047_36.0.tif Frame alignment results: distance raw = 24.8 smoothed = 21.6 Weighted resid mean = 0.12 max max = 0.21 Mean unweighted max = 0.29 Record image shifted -0.003, 0.015 um Tracking reference shifted by 0.3,-11.4 pixels, -0.003,0.015 um Saved Z = 47 at 23.19 min Tilt = 36.00 Mean = 43 Exp = 1.744 X/Y/Z = -0.633 0.665 2.769 114155.625: Saved state 47 at 36.0: def 2.669 IS 0.224 0.928 int 0.12073 exp 1.744 114155.640: SetIntensity called with factor 1.0094 114155.656: Changing (ideal) Record exposure by 1.0094 to 1.7600 114155.672: Frame time for constraint is 0.1598 114155.687: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 114159.406: recentlyReversed: dir 1 ang 37.999845 revtilt 41.001477 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.013 0.064 um, 0.9 4.6 % Error in Z Prediction -0.002 Predicted X = -0.640 StdErr = 0.024 nFit = 3 nDrop = 0 Predicted Y = 0.718 StdErr = 0.046 nFit = 5 nDrop = 0 Predicted Z = 2.654 StdErr = 0.039 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Trimming A by 20 25, reference by 20 20 Tracking image shifted -0.008, -0.011 um; position -0.632, 0.729 Measured defocus = -0.09 microns changed by -0.01 to target Trimming A by 20 25, reference by 20 20 Tracking image shifted -0.002, -0.002 um; position -0.630, 0.731 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0048_38.0.tif Frame alignment results: distance raw = 25.9 smoothed = 23.0 Weighted resid mean = 0.12 max max = 0.19 Mean unweighted max = 0.35 Record image shifted -0.004, 0.067 um Tracking reference shifted by -6.3,-49.4 pixels, -0.004,0.067 um Saved Z = 48 at 23.73 min Tilt = 38.00 Mean = 43 Exp = 1.760 X/Y/Z = -0.626 0.664 2.645 114187.609: Saved state 48 at 38.0: def 2.545 IS 0.219 0.924 int 0.12073 exp 1.760 114187.687: Restored state 46 at -58.0 for tiltInd 49 def 6.334 IS 0.914 -0.820 114187.703: SetIntensity called with factor 1.0102 114187.719: Changing (ideal) Record exposure by 1.0102 to 1.7790 114187.734: Frame time for constraint is 0.1598 114187.750: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.014 -0.055 um, 1.0 3.9 % Error in Z Prediction -0.010 Predicted X = -0.432 StdErr = 0.023 nFit = 3 nDrop = 0 Predicted Y = -1.141 StdErr = 0.034 nFit = 6 nDrop = 0 Predicted Z = 6.483 StdErr = 0.037 nFit = 4 nDrop = 0 Need regular tracking - Tracking image shifted 0.010, 0.035 um; position -0.442, -1.176 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0049_-60.0.tif Frame alignment results: distance raw = 26.8 smoothed = 27.2 Weighted resid mean = 0.14 max max = 0.22 Mean unweighted max = 0.49 Record image shifted -0.002, 0.029 um Tracking reference shifted by -2.7,-21.4 pixels, -0.002,0.029 um Saved Z = 49 at 24.23 min Tilt = -59.98 Mean = 46 Exp = 1.779 X/Y/Z = -0.440 -1.205 6.483 114218.078: Saved state 49 at -60.0: def 6.383 IS 0.947 -0.827 int 0.12073 exp 1.779 114218.094: SetIntensity called with factor 1.0109 114218.109: Changing (ideal) Record exposure by 1.0109 to 1.7984 114218.125: Frame time for constraint is 0.1598 114218.140: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction -0.008 -0.064 um, 0.6 4.6 % Predicted X = -0.446 StdErr = 0.021 nFit = 3 nDrop = 0 Predicted Y = -1.202 StdErr = 0.037 nFit = 6 nDrop = 0 Predicted Z = 6.546 StdErr = 0.041 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted -0.017, 0.092 um; position -0.430, -1.294 Measured defocus = -0.57 microns changed by 0.47 to target Trimming A by 21 120, reference by 20 20 Tracking image shifted 0.001, -0.004 um; position -0.431, -1.290 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0050_-62.0.tif Frame alignment results: distance raw = 84.4 smoothed = 66.7 Weighted resid mean = 2.04 max max = 4.21 Mean unweighted max = 45.91 Trimming A by 95 136, reference by 21 21 Record image shifted 0.922, -0.260 um Tracking reference shifted by -634.2,315.2 pixels, 0.922,-0.260 um Getting a new tracking reference after Record to reduce reference conflicts Saved Z = 50 at 24.73 min Tilt = -61.98 Mean = 33 Exp = 1.798 X/Y/Z = -1.353 -1.030 7.028 114257.250: Saved state 50 at -62.0: def 6.928 IS 1.643 -0.180 int 0.12073 exp 1.798 114257.344: Restored state 48 at 38.0 for tiltInd 51 def 2.545 IS 0.219 0.924 114257.359: SetIntensity called with factor 1.0101 114257.375: Changing (ideal) Record exposure by 1.0101 to 1.7778 114257.390: Frame time for constraint is 0.1598 114257.406: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 114266.906: recentlyReversed: dir 1 ang 40.000099 revtilt 43.000232 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.907 0.173 um, 65.5 12.5 % Error in Z Prediction 0.482 Predicted X = -0.643 StdErr = 0.027 nFit = 3 nDrop = 0 Predicted Y = 0.694 StdErr = 0.056 nFit = 5 nDrop = 0 Predicted Z = 2.551 StdErr = 0.031 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus Trimming A by 20 20, reference by 25 25 Tracking image shifted 0.021, -0.089 um; position -0.664, 0.783 Measured defocus = -0.02 microns changed by -0.08 to target Trimming A by 20 20, reference by 25 25 Tracking image shifted -0.007, 0.066 um; position -0.656, 0.717 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0051_40.0.tif Frame alignment results: distance raw = 27.6 smoothed = 24.3 Weighted resid mean = 0.14 max max = 0.23 Mean unweighted max = 0.37 Record image shifted -0.003, 0.014 um Tracking reference shifted by 0.3,-10.9 pixels, -0.003,0.014 um Saved Z = 51 at 25.48 min Tilt = 40.00 Mean = 42 Exp = 1.778 X/Y/Z = -0.653 0.702 2.528 114292.828: Saved state 51 at 40.0: def 2.428 IS 0.224 0.973 int 0.12073 exp 1.778 114292.859: SetIntensity called with factor 1.0108 114292.875: Changing (ideal) Record exposure by 1.0108 to 1.7970 114292.890: Frame time for constraint is 0.1598 114292.906: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 114296.562: recentlyReversed: dir 1 ang 41.998855 revtilt 44.995487 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.010 0.009 um, 0.7 0.6 % Error in Z Prediction -0.023 Predicted X = -0.662 StdErr = 0.046 nFit = 4 nDrop = 0 Predicted Y = 0.727 StdErr = 0.040 nFit = 6 nDrop = 0 Predicted Z = 2.402 StdErr = 0.006 nFit = 4 nDrop = 1 Need regular tracking - Tracking image shifted -0.015, 0.014 um; position -0.648, 0.713 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0052_42.0.tif Frame alignment results: distance raw = 26.6 smoothed = 25.2 Weighted resid mean = 0.14 max max = 0.22 Mean unweighted max = 0.54 Record image shifted 0.000, 0.011 um Tracking reference shifted by -1.7,-8.0 pixels, 0.000,0.011 um Saved Z = 52 at 25.92 min Tilt = 42.00 Mean = 41 Exp = 1.797 X/Y/Z = -0.648 0.702 2.402 114319.265: Saved state 52 at 42.0: def 2.302 IS 0.219 0.969 int 0.12073 exp 1.797 114319.359: Restored state 50 at -62.0 for tiltInd 53 def 6.928 IS 1.643 -0.180 114319.375: SetIntensity called with factor 1.0117 114319.390: Changing (ideal) Record exposure by 1.0117 to 1.8196 114319.406: Frame time for constraint is 0.1598 114319.422: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) Errors in X/Y Prediction 0.014 -0.025 um, 1.0 1.8 % Predicted X = -2.510 StdErr = 0.568 nFit = 4 nDrop = 0 Predicted Y = -0.923 StdErr = 0.134 nFit = 5 nDrop = 0 Predicted Z = 7.030 StdErr = 0.255 nFit = 4 nDrop = 0 Need regular tracking - Need autofocus 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. 5 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 114340.390: Restored state 52 at 42.0 for tiltInd 55 def 2.302 IS 0.219 0.969 114340.406: SetIntensity called with factor 1.0117 114340.422: Changing (ideal) Record exposure by 1.0117 to 1.8180 114340.437: Frame time for constraint is 0.1598 114340.453: Will be constrained to 1.7577 (frame 0.1598, constraint 0.1598) 114349.719: recentlyReversed: dir 1 ang 43.999110 revtilt 46.998742 back 3.000000 3mte 0.300000 Errors in X/Y Prediction 1.862 1.625 um, 134.5 117.4 % Need regular tracking - Need autofocus Tracking image shifted 0.049, -0.166 um; position -0.697, 0.868 Measured defocus = 0.22 microns changed by -0.32 to target Tracking image shifted -0.003, 0.003 um; position -0.694, 0.865 11 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0053_44.0.tif Frame alignment results: distance raw = 24.6 smoothed = 21.9 Weighted resid mean = 0.17 max max = 0.25 Mean unweighted max = 0.35 Record image shifted -0.009, 0.097 um Tracking reference shifted by -6.4,-71.4 pixels, -0.009,0.097 um Saved Z = 53 at 26.86 min Tilt = 44.00 Mean = 40 Exp = 1.818 X/Y/Z = -0.685 0.768 2.081 114375.672: Saved state 53 at 44.0: def 1.981 IS 0.219 1.048 int 0.12073 exp 1.818 114375.687: SetIntensity called with factor 1.0125 114375.703: Changing (ideal) Record exposure by 1.0125 to 1.8408 114375.719: Frame time for constraint is 0.1598 114375.734: Will be constrained to 1.9174 (frame 0.1598, constraint 0.1598) 114379.422: recentlyReversed: dir 1 ang 45.999864 revtilt 48.995997 back 3.000000 3mte 0.300000 Need regular tracking - Need autofocus Tracking image shifted -0.003, -0.056 um; position -0.682, 0.824 Measured defocus = 0.03 microns changed by -0.13 to target Tracking image shifted -0.002, -0.001 um; position -0.680, 0.825 12 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0054_46.0.tif Frame alignment results: distance raw = 28.6 smoothed = 26.7 Weighted resid mean = 0.16 max max = 0.24 Mean unweighted max = 0.48 Record image shifted -0.005, 0.068 um Tracking reference shifted by -5.9,-50.1 pixels, -0.005,0.068 um Saved Z = 54 at 27.39 min Tilt = 46.00 Mean = 38 Exp = 1.841 X/Y/Z = -0.675 0.757 1.952 114407.562: Saved state 54 at 46.0: def 1.852 IS 0.216 1.033 int 0.12073 exp 1.841 114407.578: SetIntensity called with factor 1.0135 114407.594: Changing (ideal) Record exposure by 1.0135 to 1.8657 114407.609: Frame time for constraint is 0.1598 114407.625: Will be constrained to 1.9174 (frame 0.1598, constraint 0.1598) 114410.922: recentlyReversed: dir 1 ang 47.999619 revtilt 50.982254 back 3.000000 3mte 0.300000 Predicted X = -0.666 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Y = 0.746 StdErr = 0.000 nFit = 2 nDrop = 0 Predicted Z = 1.824 StdErr = 0.000 nFit = 2 nDrop = 0 Need regular tracking - Need autofocus Tracking image shifted 0.017, -0.080 um; position -0.683, 0.826 Measured defocus = -0.05 microns changed by -0.05 to target Tracking image shifted -0.002, -0.003 um; position -0.681, 0.829 12 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0055_48.0.tif Frame alignment results: distance raw = 29.0 smoothed = 27.0 Weighted resid mean = 0.18 max max = 0.30 Mean unweighted max = 0.46 Record image shifted -0.006, 0.072 um Tracking reference shifted by -5.2,-53.2 pixels, -0.006,0.072 um Saved Z = 55 at 27.92 min Tilt = 48.00 Mean = 37 Exp = 1.866 X/Y/Z = -0.674 0.757 1.771 114439.234: Saved state 55 at 48.0: def 1.671 IS 0.216 1.032 int 0.12073 exp 1.866 114439.250: SetIntensity called with factor 1.0146 114439.265: Changing (ideal) Record exposure by 1.0146 to 1.8930 114439.281: Frame time for constraint is 0.1598 114439.297: Will be constrained to 1.9174 (frame 0.1598, constraint 0.1598) 114442.969: recentlyReversed: dir 1 ang 49.999374 revtilt 52.999507 back 3.000000 3mte 0.300000 Errors in X/Y Prediction -0.009 0.011 um, 0.6 0.8 % Error in Z Prediction -0.053 Predicted X = -0.668 StdErr = 0.006 nFit = 3 nDrop = 0 Predicted Y = 0.749 StdErr = 0.008 nFit = 3 nDrop = 0 Predicted Z = 1.624 StdErr = 0.040 nFit = 3 nDrop = 0 Need regular tracking - Tracking image shifted 0.013, -0.060 um; position -0.681, 0.809 12 frames were saved to W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075_0056_50.0.tif Frame alignment results: distance raw = 26.6 smoothed = 23.9 Weighted resid mean = 0.17 max max = 0.27 Mean unweighted max = 0.56 Record image shifted -0.007, 0.071 um Tracking reference shifted by -4.5,-52.9 pixels, -0.007,0.071 um Saved Z = 56 at 28.36 min Tilt = 50.00 Mean = 35 Exp = 1.893 X/Y/Z = -0.674 0.737 1.624 2021/11/19 22:44 W:\Zhiyang\Krios\20211117\Y00000Lamella04\20211117_075.mrc 0 57 11520 8184 10747 -10.0 50.0 2.00 REG 28.4 1x1 Continuum_K3 26000 1 0.17 LD-DS 20.0 0.0 Tilt series acquired at item # 39 with label 59 (item at 438.17, -20.54, TS at 438.28, -18.82) ################# 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