Multiprocessor-enabled deconvolution (using 8 processors) decon3d: 3d deconvolution... Transferring field 0: CameraType c 30 CoolSNAP_HQ2 / HQ2-ICX285 Transferring field 1: CameraGain f 8 1.000000 Transferring field 2: CameraBinning i 1 1 ChI Sel? Proc? WPass WavI WavO 0 0 1 0 457 457 1 1 1 0 617 617 2 2 1 0 525 525 3 0 0 0 0 0 4 0 0 0 0 0 5 0 0 0 0 0 6 0 0 0 0 137345151936011173888 7 0 0 0 0 0 8 0 0 0 137664696403321946112 137664696403321946112 9 0 0 0 0 0 10 0 0 0 0 0 11 0 0 0 0 0 Passed camera intensity offset 100.000000 is greater than min intensity 89.000000... Resetting all channels to default offset. Image Correction Input image file............................. /data1/Stojic_lab/Claudia_Kutter/09Nov2021_cells/A375_linc326_002_01_R3D.dv Output image file............................ /tmp/A375_linc326_002_01_R3D_D3D.dv_cor_QGM Polynomial order............................. 5 X min and max for normalization.............. 0 895 Y min and max for normalization.............. 0 895 Normalization................................ on Type of normalization........................ 1 Camera Intensity Offset (bias)............... 0.00 Bleach correction............................ on Z-line correction............................ on Intensity offset(s).......................... 0.00 50.00 50.00 Photosensor reference value(s)............... 2.6660e+03 5.6417e+05 6.1502e+05 Correcting time point 1, wavelength 1, panel 1... Reading 3D image data (37 sections)... Photobleaching correction disabled. Best-fit slope is <0 (-1.00862e-04). Z Ref Scaled Ref Photosensor Scaled Photosensor - --- ---------- ----------- ------------------ 1 116.29 116.29 2666.0 2666.0 1.0000/1.0000 2 116.32 116.28 2283.0 2282.2 1.1678/0.9997 3 116.51 116.26 2351.0 2346.0 1.1340/0.9979 4 116.73 116.25 2723.0 2711.8 0.9791/0.9959 5 116.91 116.24 2824.0 2807.8 0.9441/0.9943 6 117.05 116.23 2431.0 2414.1 1.0967/0.9930 7 117.10 116.22 2267.0 2249.9 1.1760/0.9925 8 117.36 116.21 2612.0 2586.3 1.0207/0.9902 9 117.64 116.19 2939.0 2902.9 0.9071/0.9877 10 117.60 116.18 2684.0 2651.6 0.9933/0.9879 11 117.75 116.17 2285.0 2254.4 1.1667/0.9866 12 117.86 116.16 2376.0 2341.6 1.1221/0.9855 13 118.00 116.15 2781.0 2737.5 0.9586/0.9843 14 118.19 116.14 2770.0 2721.8 0.9625/0.9826 15 118.15 116.12 2421.0 2379.4 1.1012/0.9828 16 118.34 116.11 2279.0 2236.0 1.1698/0.9811 17 118.38 116.10 2649.0 2598.1 1.0064/0.9808 18 118.66 116.09 2927.0 2863.5 0.9108/0.9783 19 118.74 116.08 2657.0 2597.4 1.0034/0.9776 20 118.91 116.07 2280.0 2225.4 1.1693/0.9761 21 119.06 116.05 2423.0 2361.9 1.1003/0.9748 22 118.99 116.04 2781.0 2712.2 0.9586/0.9753 23 119.20 116.03 2732.0 2659.3 0.9758/0.9734 24 119.23 116.02 2354.0 2290.5 1.1325/0.9730 25 119.42 116.01 2275.0 2210.1 1.1719/0.9715 26 119.45 116.00 2671.0 2593.7 0.9981/0.9711 27 119.39 115.98 2928.0 2844.5 0.9105/0.9715 28 119.68 115.97 2604.0 2523.3 1.0238/0.9690 29 119.72 115.96 2278.0 2206.5 1.1703/0.9686 30 119.71 115.95 2457.0 2379.9 1.0851/0.9686 31 119.82 115.94 2812.0 2720.9 0.9481/0.9676 32 119.90 115.93 2731.0 2640.4 0.9762/0.9668 33 119.86 115.91 2321.0 2244.7 1.1486/0.9671 34 120.03 115.90 2269.0 2190.9 1.1750/0.9656 35 120.14 115.89 2683.0 2588.1 0.9937/0.9646 36 120.21 115.88 2916.0 2810.9 0.9143/0.9640 37 120.24 115.87 2577.0 2483.2 1.0345/0.9636 Correcting time point 1, wavelength 2, panel 1... Reading 3D image data (37 sections)... Z Ref Scaled Ref Photosensor Scaled Photosensor - --- ---------- ----------- ------------------ 1 114.59 114.59 564171.0 564171.0 1.0000/1.0000 2 114.53 114.59 562601.0 562902.8 1.0028/1.0005 3 114.47 114.59 563681.0 564278.2 1.0009/1.0011 4 114.46 114.59 564602.0 565247.1 0.9992/1.0011 5 114.38 114.59 563886.0 564888.4 1.0005/1.0018 6 114.33 114.59 563018.0 564291.7 1.0020/1.0023 7 114.36 114.59 564431.0 565533.9 0.9995/1.0020 8 114.28 114.59 563645.0 565164.6 1.0009/1.0027 9 114.27 114.59 564192.0 565779.1 1.0000/1.0028 10 114.27 114.59 564019.0 565572.6 1.0003/1.0028 11 114.25 114.59 563650.0 565295.6 1.0009/1.0029 12 114.24 114.59 563583.0 565320.8 1.0010/1.0031 13 114.21 114.59 563608.0 565452.8 1.0010/1.0033 14 114.24 114.59 565719.0 567443.1 0.9973/1.0030 15 114.19 114.59 564025.0 565977.4 1.0003/1.0035 16 114.16 114.59 563391.0 565502.4 1.0014/1.0037 17 114.17 114.59 563435.0 565496.4 1.0013/1.0037 18 114.20 114.59 564411.0 566335.8 0.9996/1.0034 19 114.14 114.59 564036.0 566224.8 1.0002/1.0039 20 114.19 114.59 564046.0 566006.2 1.0002/1.0035 21 114.19 114.59 564392.0 566351.2 0.9996/1.0035 22 114.14 114.59 563784.0 566013.6 1.0007/1.0040 23 114.14 114.59 564178.0 566413.5 1.0000/1.0040 24 114.10 114.59 563427.0 565815.9 1.0013/1.0042 25 114.21 114.59 564489.0 566343.1 0.9994/1.0033 26 114.13 114.59 564651.0 566938.9 0.9991/1.0041 27 114.18 114.59 564573.0 566606.6 0.9993/1.0036 28 114.17 114.59 563165.0 565244.8 1.0018/1.0037 29 114.17 114.59 563826.0 565901.6 1.0006/1.0037 30 114.23 114.59 563825.0 565588.1 1.0006/1.0031 31 114.22 114.59 563637.0 565468.6 1.0009/1.0032 32 114.29 114.59 564553.0 566047.0 0.9993/1.0026 33 114.24 114.59 563381.0 565113.2 1.0014/1.0031 34 114.22 114.59 562981.0 564787.7 1.0021/1.0032 35 114.22 114.59 562629.0 564459.0 1.0027/1.0033 36 114.26 114.59 563540.0 565133.4 1.0011/1.0028 37 114.27 114.59 564068.0 565654.3 1.0002/1.0028 Correcting time point 1, wavelength 3, panel 1... Reading 3D image data (37 sections)... Photobleaching correction disabled. Best-fit slope is <0 (-1.04468e-06). Z Ref Scaled Ref Photosensor Scaled Photosensor - --- ---------- ----------- ------------------ 1 104.43 104.43 615019.0 615019.0 1.0000/1.0000 2 104.30 104.43 615200.0 616010.4 0.9997/1.0013 3 104.23 104.43 617558.0 618758.1 0.9959/1.0019 4 104.15 104.43 615247.0 616933.5 0.9996/1.0027 5 104.14 104.43 615297.0 617032.1 0.9995/1.0028 6 104.15 104.43 618778.0 620434.3 0.9939/1.0027 7 104.14 104.43 619262.0 620991.6 0.9931/1.0028 8 104.10 104.43 616420.0 618368.6 0.9977/1.0032 9 104.11 104.43 615662.0 617582.7 0.9990/1.0031 10 104.11 104.43 611982.0 613902.2 1.0050/1.0031 11 104.12 104.43 614984.0 616831.0 1.0001/1.0030 12 104.11 104.43 616718.0 618602.4 0.9972/1.0031 13 104.12 104.43 615364.0 617207.8 0.9994/1.0030 14 104.14 104.43 615658.0 617405.8 0.9990/1.0028 15 104.14 104.43 612977.0 614717.7 1.0033/1.0028 16 104.15 104.43 614005.0 615692.8 1.0017/1.0027 17 104.22 104.43 617593.0 618872.4 0.9958/1.0021 18 104.16 104.43 615709.0 617315.1 0.9989/1.0026 19 104.22 104.43 618353.0 619598.3 0.9946/1.0020 20 104.23 104.43 616026.0 617230.4 0.9984/1.0020 21 104.24 104.43 617588.0 618694.4 0.9958/1.0018 22 104.26 104.43 616879.0 617909.7 0.9970/1.0017 23 104.29 104.43 620053.0 620907.6 0.9919/1.0014 24 104.28 104.43 616202.0 617107.6 0.9981/1.0015 25 104.29 104.43 616312.0 617171.4 0.9979/1.0014 26 104.29 104.43 617231.0 618082.4 0.9964/1.0014 27 104.32 104.43 616697.0 617350.4 0.9973/1.0011 28 104.35 104.43 618765.0 619268.9 0.9939/1.0008 29 104.34 104.43 614488.0 615051.9 1.0009/1.0009 30 104.36 104.43 615861.0 616258.7 0.9986/1.0006 31 104.38 104.43 613590.0 613889.3 1.0023/1.0005 32 104.40 104.43 619323.0 619482.7 0.9931/1.0003 33 104.44 104.43 614563.0 614516.9 1.0007/0.9999 34 104.42 104.43 612997.0 613085.8 1.0033/1.0001 35 104.45 104.43 616454.0 616310.2 0.9977/0.9998 36 104.45 104.43 617011.0 616867.2 0.9968/0.9998 37 104.50 104.43 617573.0 617129.4 0.9959/0.9993 ---------------------------------------------------- Deconvolution Results: Transferring field 0: CameraType c 30 CoolSNAP_HQ2 / HQ2-ICX285 Transferring field 1: CameraGain f 8 1.000000 Transferring field 2: CameraBinning i 1 1 Transferring field 3: COR_BlchFitSlp f 8 0.000001 Transferring field 4: COR_BlchFitInt f 8 4.738676 Transferring field 5: COR_BlchFitChi f 8 0.000000 Opening OTF file (/usr/local/otf/Olympus_100X_140_10002.otf)... decon3d: 3D iterative constrained deconvolution Input image file............................. /tmp/A375_linc326_002_01_R3D_D3D.dv_cor_QGM Output image file............................ /data1/Stojic_lab/Claudia_Kutter/09Nov2021_cells/A375_linc326_002_01_R3D_D3D.dv Method (1=add,2=ratio,3=enh add,4=enh ratio). 4 Values subtracted from image................. 0.00 0.00 0.00 Intensity scaling factor..................... 1.0000 Border roll-off size (pixels)................ 28 Pre-filter cut-off values (microns).......... 0.2000, 0.0500 Sigma for smoothing.......................... 0.3050 Sigma for smoothing Wiener filter............ 0.1100 S:N value for Wiener filter.................. 0.1010 Z padding.................................... 128 Number of iterations before smoothing........ 5 Maximum number of iterations................. 10 Autoscaling.................................. enabled Allocating image memory (nx=896,ny=896,sOpt->nzo=37,total=28.3 MP)... Generating pre-filter and smoothing function... Wavelength 457.0 nm processed with OTF measured at 617.0 nm (scale=1.350)... Preparing OTF (128 128 129)... Deconvolving wavelength 457.0, time point 1 (3D)... Z noise filter threshold=1096.00 Prefiltering wavelength 457.0... decon3d: starting iterations... Standard Avg Counts Iteration Residual (R) Residual Normalized R --------- ------------ -------- ------------ 1 0.01582844 1.83 1.000 2 0.01058190 1.22 0.669 3 0.01032698 1.19 0.652 4 0.01020181 1.18 0.645 5 0.01009742 1.17 0.638 6 0.01017369 1.18 0.643 7 0.01005947 1.16 0.636 8 0.00996795 1.15 0.630 9 0.00988646 1.14 0.625 10 0.00981340 1.13 0.620 decon3d: finished iterations (10/10). Wavelength 617.0 nm processed with OTF measured at 617.0 nm (scale=1.000)... Preparing OTF (128 128 129)... Deconvolving wavelength 617.0, time point 1 (3D)... Z noise filter threshold=1112.00 Prefiltering wavelength 617.0... decon3d: starting iterations... Standard Avg Counts Iteration Residual (R) Residual Normalized R --------- ------------ -------- ------------ 1 0.01098271 1.25 1.000 2 0.01066630 1.22 0.971 3 0.01053426 1.20 0.959 4 0.01044607 1.19 0.951 5 0.01040727 1.19 0.948 6 0.01044292 1.19 0.951 7 0.01039410 1.18 0.946 8 0.01035965 1.18 0.943 9 0.01033002 1.18 0.941 10 0.01030481 1.17 0.938 decon3d: finished iterations (10/10). Wavelength 525.0 nm processed with OTF measured at 617.0 nm (scale=1.175)... Preparing OTF (128 128 129)... Deconvolving wavelength 525.0, time point 1 (3D)... Z noise filter threshold=1104.00 Prefiltering wavelength 525.0... decon3d: starting iterations... Standard Avg Counts Iteration Residual (R) Residual Normalized R --------- ------------ -------- ------------ 1 0.01043341 1.08 1.000 2 0.01018394 1.06 0.976 3 0.01005103 1.04 0.963 4 0.00995878 1.03 0.955 5 0.00989701 1.03 0.949 6 0.00994277 1.03 0.953 7 0.00987491 1.03 0.946 8 0.00982031 1.02 0.941 9 0.00977147 1.01 0.937 10 0.00972786 1.01 0.932 decon3d: finished iterations (10/10). Total deconvolution time: 00:00:19