I wrote earlier today suggesting that stitching the microscope images using the translation model might require assigning a small field of view to the individual images. I'm going to backtrack on that. I have realised that I had failed to notice that the projection was set at equirectangular. With a small field of view for the individual images there was no noticeable difference between these two settings, but when the fov was large it showed. When I corrected that setting for the translation model it came out fine with an individual fov of 50 degrees. So I am back to thinking that for the translation model the fov of the individual images really isn't all that important.

For the small-angle optimisation, of course, it does matter that the angle is at least small, and that approach remains sensible, it seems to me.

I attach a pto file for the 50-degree case and, for the record, a revised file for the small-angle approach.

Roger Broadie

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# hugin project file
#hugin_ptoversion 2
p f0 w3842 h2049 v1.5  E0 R0 S407,3842,82,1230 n"TIFF_m c:LZW r:CROP"
m g1 i0 f0 m2 p0.00784314

# image lines
#-hugin  cropFactor=1
i w1280 h960 f0 v0.498804477515136 Ra0 Rb0 Rc0 Rd0 Re0 Eev0 Er1 Eb1 r0 p0 y0 
TrX0 TrY0 TrZ0 j0 a-0.000445623794635797 b0.00221343669767907 c0 
d-251.254902657968 e373.096387537286 g0 t0 Va1 Vb0 Vc0 Vd0 Vx0 Vy0  Vm5 n"C.jpg"
#-hugin  cropFactor=1
i w1280 h960 f0 v=0 Ra=0 Rb=0 Rc=0 Rd=0 Re=0 Eev=0 Er=0 Eb=0 
r-0.0219186169053622 p-0.0277330653435229 y-0.411778618135149 TrX0 TrY0 TrZ0 j0 
a=0 b=0 c=0 d=0 e=0 g=0 t=0 Va=0 Vb=0 Vc=0 Vd=0 Vx=0 Vy=0  Vm5 n"L.jpg"
#-hugin  cropFactor=1
i w1280 h960 f0 v=0 Ra=0 Rb=0 Rc=0 Rd=0 Re=0 Eev=0 Er=0 Eb=0 r0.119939475544224 
p0.0255943287070798 y0.390460103516758 TrX0 TrY0 TrZ0 j0 a=0 b=0 c=0 d=0 e=0 
g=0 t=0 Va=0 Vb=0 Vc=0 Vd=0 Vx=0 Vy=0  Vm5 n"R.jpg"


# specify variables that should be optimized
v Ra0
v Rb0
v Rc0
v Rd0
v Re0
v Vb0
v Vc0
v Vd0
v r1
v p1
v y1
v r2
v p2
v y2
v


# control points
c n0 N1 x121.366605372901 y266.875221082265 X1182.95364045297 Y194.960627931095 
t0
c n0 N1 x101.229767933888 y456.460674367923 X1158.12768842316 Y384.181099869508 
t0
c n0 N1 x55.1565717612281 y638.5852324626 X1113.66140708987 Y567.226377378495 t0
c n0 N1 x116.445220128595 y770.769177862684 X1175.20099412598 Y698.27896268101 
t0
c n0 N2 x1089.97524640569 y153.240412023151 X86.3669838157867 Y222.062429112849 
t0
c n0 N2 x1088.59641675203 y336.172577963678 X82.3844287721356 Y403.014890074045 
t0
c n0 N2 x1084.24353796175 y359.622103527515 X78.4734027646847 Y426.556905109347 
t0
c n0 N2 x1096.00073725322 y73.6457285046083 X88.4777882104511 Y141.838449886841 
t0
c n0 N2 x1107.70561550204 y295.537688829494 X101.76894059982 Y363.247985098843 
t0
c n0 N2 x1112.96445573566 y354.583587651547 X108.405762265297 Y422.541427615125 
t0
c n0 N2 x1140.14969151846 y136.599461656704 X133.227480168959 Y204.344774809217 
t0
c n0 N2 x1132.62584592775 y351.317619264639 X127.134917878432 Y420.205898375112 
t0
c n0 N2 x1138.08801713661 y361.994060501543 X133.793656414132 Y429.908683111741 
t0
c n0 N2 x1163.36212949881 y777.76555080603 X160.343130880497 Y842.99699997452 t0

#hugin_optimizeReferenceImage 0
#hugin_blender enblend
#hugin_remapper nona
#hugin_enblendOptions 
#hugin_enfuseOptions 
#hugin_hdrmergeOptions -m avg -c
#hugin_outputLDRBlended true
#hugin_outputLDRLayers false
#hugin_outputLDRExposureRemapped false
#hugin_outputLDRExposureLayers false
#hugin_outputLDRExposureBlended false
#hugin_outputLDRStacks false
#hugin_outputLDRExposureLayersFused false
#hugin_outputHDRBlended false
#hugin_outputHDRLayers false
#hugin_outputHDRStacks false
#hugin_outputLayersCompression LZW
#hugin_outputImageType tif
#hugin_outputImageTypeCompression LZW
#hugin_outputJPEGQuality 80
#hugin_outputImageTypeHDR exr
#hugin_outputImageTypeHDRCompression LZW
#hugin_outputStacksMinOverlap 0.7
#hugin_outputLayersExposureDiff 0.5
#hugin_optimizerMasterSwitch 0
#hugin_optimizerPhotoMasterSwitch 21

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# hugin project file
#hugin_ptoversion 2
p f0 w3488 h1479 v104  E0 R0 S54,3399,182,1310 n"TIFF_m c:LZW r:CROP"
m g1 i0 f0 m2 p0.00784314

# image lines
#-hugin  cropFactor=1
i w1280 h960 f0 v49.9999907314703 Ra0 Rb0 Rc0 Rd0 Re0 Eev0 Er1 Eb1 
r-0.043073508005528 p-0.277931192703748 y0.158477699676188 TrX0 TrY0 TrZ0 j0 
a-0.00786401431912753 b0.0150809539022988 c0 d0 e0 g0 t0 Va1 Vb0 Vc0 Vd0 Vx0 
Vy0  Vm5 n"C.jpg"
#-hugin  cropFactor=1
i w1280 h960 f0 v=0 Ra=0 Rb=0 Rc=0 Rd=0 Re=0 Eev=0 Er=0 Eb=0 
r-0.0130553005610733 p-0.331678301748495 y0.0355303000911981 
TrX-0.768297744613223 TrY0.051783579487861 TrZ-0.000856563750715794 j0 a=0 b=0 
c=0 d=0 e=0 g=0 t=0 Va=0 Vb=0 Vc=0 Vd=0 Vx=0 Vy=0  Vm5 n"L.jpg"
#-hugin  cropFactor=1
i w1280 h960 f0 v=0 Ra=0 Rb=0 Rc=0 Rd=0 Re=0 Eev=0 Er=0 Eb=0 
r0.0143792965052305 p-0.109333735652189 y0.0842215852251854 
TrX0.735142099066223 TrY-0.0461959504548703 TrZ0.00509861059301879 j0 a=0 b=0 
c=0 d=0 e=0 g=0 t=0 Va=0 Vb=0 Vc=0 Vd=0 Vx=0 Vy=0  Vm5 n"R.jpg"


# specify variables that should be optimized
v Ra0
v Rb0
v Rc0
v Rd0
v Re0
v r0
v p0
v y0
v a0
v b0
v Vb0
v Vc0
v Vd0
v r1
v p1
v y1
v TrX1
v TrY1
v TrZ1
v r2
v p2
v y2
v TrX2
v TrY2
v TrZ2
v


# control points
c n0 N1 x121.366605372901 y266.875221082265 X1182.95364045297 Y194.960627931095 
t0
c n0 N1 x101.229767933888 y456.460674367923 X1158.12768842316 Y384.181099869508 
t0
c n0 N1 x55.1565717612281 y638.5852324626 X1113.66140708987 Y567.226377378495 t0
c n0 N1 x116.445220128595 y770.769177862684 X1175.20099412598 Y698.27896268101 
t0
c n0 N2 x1089.97524640569 y153.240412023151 X86.3669838157867 Y222.062429112849 
t0
c n0 N2 x1088.59641675203 y336.172577963678 X82.3844287721356 Y403.014890074045 
t0
c n0 N2 x1084.24353796175 y359.622103527515 X78.4734027646847 Y426.556905109347 
t0
c n0 N2 x1096.00073725322 y73.6457285046083 X88.4777882104511 Y141.838449886841 
t0
c n0 N2 x1107.70561550204 y295.537688829494 X101.76894059982 Y363.247985098843 
t0
c n0 N2 x1112.96445573566 y354.583587651547 X108.405762265297 Y422.541427615125 
t0
c n0 N2 x1140.14969151846 y136.599461656704 X133.227480168959 Y204.344774809217 
t0
c n0 N2 x1132.62584592775 y351.317619264639 X127.134917878432 Y420.205898375112 
t0
c n0 N2 x1138.08801713661 y361.994060501543 X133.793656414132 Y429.908683111741 
t0
c n0 N2 x1163.36212949881 y777.76555080603 X160.343130880497 Y842.99699997452 t0
c n0 N0 x110.769230769231 y6.15384615384615 X1168 Y8 t2
c n0 N0 x97 y958 X1155 Y958 t2
c n0 N0 x0 y857 X0 Y25 t1
c n0 N0 x634 y903 X633 Y28 t1
c n1 N1 x147.692307692308 y0 X1077 Y0 t2
c n1 N1 x59 y959 X1187 Y959 t2
c n1 N1 x1276 y17 X1274 Y886 t1
c n1 N1 x2 y942 X0 Y25 t1
c n2 N2 x86 y959 X1071 Y959 t2
c n2 N2 x43 y3 X1194 Y3 t2
c n2 N2 x0 y34 X0 Y931 t1
c n2 N2 x1278 y54 X1278 Y888 t1

# masks
k i0 t1 p"-9 -26 315 -26 292 371 270 752 -35 983 -35 227"
k i2 t1 p"600 -63 664 996 -43 1006 -43 -43"

#hugin_optimizeReferenceImage 0
#hugin_blender enblend
#hugin_remapper nona
#hugin_enblendOptions 
#hugin_enfuseOptions 
#hugin_hdrmergeOptions -m avg -c
#hugin_outputLDRBlended true
#hugin_outputLDRLayers false
#hugin_outputLDRExposureRemapped false
#hugin_outputLDRExposureLayers false
#hugin_outputLDRExposureBlended false
#hugin_outputLDRStacks false
#hugin_outputLDRExposureLayersFused false
#hugin_outputHDRBlended false
#hugin_outputHDRLayers false
#hugin_outputHDRStacks false
#hugin_outputLayersCompression LZW
#hugin_outputImageType tif
#hugin_outputImageTypeCompression LZW
#hugin_outputJPEGQuality 80
#hugin_outputImageTypeHDR exr
#hugin_outputImageTypeHDRCompression LZW
#hugin_outputStacksMinOverlap 0.7
#hugin_outputLayersExposureDiff 0.5
#hugin_optimizerMasterSwitch 0
#hugin_optimizerPhotoMasterSwitch 21

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A list of frequently asked questions is available at: 
http://wiki.panotools.org/Hugin_FAQ
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