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 0 7 0 0 0 0 0 8 0 0 0 0 0 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 91.000000... Resetting all channels to default offset. Image Correction Input image file............................. /data1/Stojic_lab/Claudia_Kutter/09Nov2021_cells/A375_linc326_002_02_R3D.dv Output image file............................ /tmp/A375_linc326_002_02_R3D_D3D.dv_cor_KBI 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)............... 2.4550e+03 5.6347e+05 6.1488e+05 Correcting time point 1, wavelength 1, panel 1... Reading 3D image data (37 sections)... Photobleaching correction disabled. Best-fit slope is <0 (-6.72337e-05). Z Ref Scaled Ref Photosensor Scaled Photosensor - --- ---------- ----------- ------------------ 1 129.38 129.38 2455.0 2455.0 1.0000/1.0000 2 129.50 129.37 2323.0 2320.7 1.0568/0.9990 3 129.75 129.36 2667.0 2658.8 0.9205/0.9969 4 130.03 129.35 2975.0 2959.4 0.8252/0.9948 5 130.23 129.34 2712.0 2693.4 0.9052/0.9931 6 130.53 129.33 2340.0 2318.5 1.0491/0.9908 7 130.70 129.32 2431.0 2405.4 1.0099/0.9895 8 130.72 129.31 2784.0 2754.0 0.8818/0.9892 9 131.25 129.31 2807.0 2765.4 0.8746/0.9852 10 131.27 129.30 2427.0 2390.5 1.0115/0.9850 11 131.53 129.29 2328.0 2288.4 1.0546/0.9830 12 131.84 129.28 2709.0 2656.5 0.9062/0.9806 13 132.14 129.27 2955.0 2890.9 0.8308/0.9783 14 132.55 129.26 2688.0 2621.3 0.9133/0.9752 15 132.62 129.25 2343.0 2283.6 1.0478/0.9747 16 133.05 129.25 2466.0 2395.4 0.9955/0.9714 17 133.14 129.24 2824.0 2741.1 0.8693/0.9707 18 133.48 129.23 2775.0 2686.6 0.8847/0.9681 19 133.68 129.22 2397.0 2316.9 1.0242/0.9666 20 134.21 129.21 2338.0 2250.9 1.0500/0.9628 21 134.40 129.20 2714.0 2609.0 0.9046/0.9613 22 134.78 129.19 2948.0 2825.7 0.8328/0.9585 23 134.69 129.18 2674.0 2564.7 0.9181/0.9591 24 135.02 129.18 2331.0 2230.1 1.0532/0.9567 25 135.33 129.17 2504.0 2390.0 0.9804/0.9545 26 135.34 129.16 2854.0 2723.6 0.8602/0.9543 27 135.90 129.15 2772.0 2634.2 0.8856/0.9503 28 136.10 129.14 2358.0 2237.5 1.0411/0.9489 29 136.65 129.13 2316.0 2188.5 1.0600/0.9450 30 136.74 129.12 2728.0 2576.1 0.8999/0.9443 31 136.74 129.12 2932.0 2768.5 0.8373/0.9442 32 137.35 129.11 2640.0 2481.5 0.9299/0.9400 33 137.36 129.10 2336.0 2195.5 1.0509/0.9398 34 138.03 129.09 2544.0 2379.2 0.9650/0.9352 35 138.14 129.08 2901.0 2710.7 0.8463/0.9344 36 138.43 129.07 2753.0 2566.9 0.8918/0.9324 37 138.80 129.06 2333.0 2169.3 1.0523/0.9298 Correcting time point 1, wavelength 2, panel 1... Reading 3D image data (37 sections)... Z Ref Scaled Ref Photosensor Scaled Photosensor - --- ---------- ----------- ------------------ 1 116.15 116.15 563472.0 563472.0 1.0000/1.0000 2 116.07 116.15 563929.0 564312.1 0.9992/1.0007 3 116.03 116.15 564352.0 564948.6 0.9984/1.0011 4 115.93 116.15 563673.0 564762.9 0.9996/1.0019 5 115.89 116.15 563537.0 564825.1 0.9999/1.0023 6 115.85 116.15 562982.0 564478.9 1.0009/1.0027 7 115.81 116.15 562398.0 564067.4 1.0019/1.0030 8 115.75 116.15 563049.0 565028.2 1.0008/1.0035 9 115.75 116.15 563414.0 565367.7 1.0001/1.0035 10 115.72 116.15 563347.0 565464.6 1.0002/1.0038 11 115.70 116.15 562375.0 564585.2 1.0020/1.0039 12 115.66 116.15 563674.0 566063.2 0.9996/1.0042 13 115.68 116.15 564608.0 566942.0 0.9980/1.0041 14 115.61 116.15 562956.0 565587.9 1.0009/1.0047 15 115.61 116.15 562283.0 564932.2 1.0021/1.0047 16 115.64 116.15 564692.0 567181.7 0.9978/1.0044 17 115.64 116.15 563964.0 566481.2 0.9991/1.0045 18 115.58 116.15 563346.0 566121.0 1.0002/1.0049 19 115.62 116.15 564601.0 567188.0 0.9980/1.0046 20 115.57 116.15 563849.0 566698.9 0.9993/1.0051 21 115.62 116.15 565249.0 567865.4 0.9969/1.0046 22 115.56 116.15 563909.0 566799.6 0.9992/1.0051 23 115.59 116.15 563591.0 566341.7 0.9998/1.0049 24 115.61 116.15 563556.0 566224.1 0.9999/1.0047 25 115.59 116.15 563720.0 566475.1 0.9996/1.0049 26 115.58 116.15 563679.0 566481.1 0.9996/1.0050 27 115.62 116.15 563761.0 566383.1 0.9995/1.0047 28 115.64 116.15 563204.0 565723.1 1.0005/1.0045 29 115.63 116.15 562943.0 565501.7 1.0009/1.0045 30 115.63 116.15 564199.0 566745.4 0.9987/1.0045 31 115.66 116.15 565072.0 567465.1 0.9972/1.0042 32 115.63 116.15 563662.0 566191.3 0.9997/1.0045 33 115.65 116.15 562902.0 565367.8 1.0010/1.0044 34 115.68 116.15 563765.0 566078.3 0.9995/1.0041 35 115.67 116.15 563706.0 566075.5 0.9996/1.0042 36 115.68 116.15 564453.0 566751.3 0.9983/1.0041 37 115.72 116.15 563598.0 565728.9 0.9998/1.0038 Correcting time point 1, wavelength 3, panel 1... Reading 3D image data (37 sections)... Photobleaching correction disabled. Best-fit slope is <0 (-8.96461e-06). Z Ref Scaled Ref Photosensor Scaled Photosensor - --- ---------- ----------- ------------------ 1 104.98 104.98 614878.0 614878.0 1.0000/1.0000 2 104.80 104.98 615781.0 616857.2 0.9985/1.0017 3 104.70 104.98 615458.0 617131.2 0.9991/1.0027 4 104.66 104.98 616771.0 618644.9 0.9969/1.0030 5 104.65 104.98 617056.0 618976.6 0.9965/1.0031 6 104.64 104.98 616676.0 618672.2 0.9971/1.0032 7 104.63 104.98 617176.0 619243.2 0.9963/1.0033 8 104.60 104.98 617176.0 619366.2 0.9963/1.0035 9 104.60 104.97 616027.0 618239.8 0.9981/1.0036 10 104.58 104.97 616059.0 618347.3 0.9981/1.0037 11 104.58 104.97 615792.0 618104.5 0.9985/1.0038 12 104.60 104.97 615124.0 617322.7 0.9996/1.0036 13 104.63 104.97 616491.0 618498.2 0.9974/1.0033 14 104.62 104.97 616315.0 618370.2 0.9977/1.0033 15 104.65 104.97 617252.0 619154.6 0.9962/1.0031 16 104.61 104.97 615854.0 617960.1 0.9984/1.0034 17 104.67 104.97 618308.0 620069.6 0.9945/1.0028 18 104.66 104.97 616297.0 618093.5 0.9977/1.0029 19 104.68 104.96 617944.0 619618.7 0.9950/1.0027 20 104.68 104.96 616546.0 618230.0 0.9973/1.0027 21 104.71 104.96 616747.0 618253.7 0.9970/1.0024 22 104.70 104.96 615832.0 617386.8 0.9985/1.0025 23 104.72 104.96 614801.0 616241.1 1.0001/1.0023 24 104.73 104.96 614497.0 615839.7 1.0006/1.0022 25 104.76 104.96 615117.0 616271.8 0.9996/1.0019 26 104.82 104.96 618076.0 618900.7 0.9948/1.0013 27 104.80 104.96 618203.0 619106.1 0.9946/1.0015 28 104.82 104.96 615383.0 616173.1 0.9992/1.0013 29 104.81 104.96 612866.0 613700.9 1.0033/1.0014 30 104.88 104.95 617144.0 617559.2 0.9963/1.0007 31 104.87 104.95 613794.0 614310.1 1.0018/1.0008 32 104.92 104.95 618902.0 619086.4 0.9935/1.0003 33 104.94 104.95 615487.0 615556.2 0.9990/1.0001 34 104.97 104.95 617529.0 617391.4 0.9957/0.9998 35 104.99 104.95 618057.0 617820.0 0.9949/0.9996 36 104.98 104.95 617308.0 617120.8 0.9961/0.9997 37 104.99 104.95 615123.0 614851.6 0.9996/0.9996 ---------------------------------------------------- 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 0.000002 Transferring field 4: COR_BlchFitInt f 8 4.750161 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_02_R3D_D3D.dv_cor_KBI Output image file............................ /data1/Stojic_lab/Claudia_Kutter/09Nov2021_cells/A375_linc326_002_02_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 0.01892635 2.43 1.000 2 0.01068144 1.37 0.564 3 0.01034290 1.33 0.546 4 0.01020378 1.31 0.539 5 0.01009561 1.30 0.533 6 0.01017712 1.31 0.538 7 0.01005761 1.29 0.531 8 0.00996279 1.28 0.526 9 0.00987880 1.27 0.522 10 0.00980370 1.26 0.518 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 0.01124920 1.30 1.000 2 0.01073719 1.24 0.954 3 0.01059482 1.22 0.942 4 0.01050362 1.21 0.934 5 0.01046434 1.21 0.930 6 0.01050029 1.21 0.933 7 0.01045081 1.21 0.929 8 0.01041576 1.20 0.926 9 0.01038561 1.20 0.923 10 0.01036000 1.20 0.921 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 0.01047682 1.09 1.000 2 0.01020337 1.06 0.974 3 0.01006894 1.05 0.961 4 0.00997584 1.04 0.952 5 0.00991369 1.03 0.946 6 0.00995935 1.04 0.951 7 0.00989147 1.03 0.944 8 0.00983651 1.03 0.939 9 0.00978738 1.02 0.934 10 0.00974349 1.02 0.930 decon3d: finished iterations (10/10). Total deconvolution time: 00:00:20