satijalab / seurat

R toolkit for single cell genomics
http://www.satijalab.org/seurat
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Issue with LoadVizgen #8597

Open angie-john opened 8 months ago

angie-john commented 8 months ago

I'm trying to follow this tutorial: https://satijalab.org/seurat/articles/seurat5_spatial_vignette_2 but I'm unable to properly load the data. I'm having the same issue as #7142 however, I'm unable to fix it like they did. I believe I have the data downloaded and named correctly. This is the error I get: image

This is my session info: image

These are what my files look like: image image

I would greatly appreciate any help with this.

alikhuseynov commented 7 months ago

I'm not from Seurat dev team.. however, It is related to future library. Try to install this PR (still under review) I did for updated Vizgen output

Here is an example on how to use that optimized function. Instead of using future, use BiocParallel and set the cores within LoadVizgen(use.BiocParallel = TRUE, workers.MulticoreParam = 4) that should work.

angie-john commented 7 months ago

I tried to do what you suggested but received these errors. I believe I had installed the packages correctly though.

vizgen error biocparallel

alikhuseynov commented 7 months ago

I tried to do what you suggested but received these errors. I believe I had installed the packages correctly though.

vizgen error biocparallel

I don't think it uses the optimized LoadVizgen since it tells you that there are unused arguments

angie-john commented 7 months ago

Hm.. do you know how I would use the optimized LoadVizgen instead?

alikhuseynov commented 7 months ago

Hm.. do you know how I would use the optimized LoadVizgen instead?

You didn't install that PR on Vizgen properly. After install restart your R session, then it should work.

angie-john commented 7 months ago

I was able to get that to work but I'm still getting the issue with the future library

error cannot ffind HDF5 file Do you have any other suggestions for what could fix it? Greatly appreciate your help so far.

groverj3 commented 7 months ago

Adding my voice to the choir of users who have run into this issue while testing Seurat's suitability for our spatial transcriptomics experiments.

I was able to get data loaded using @alikhuseynov 's PR. Hopefully this is incorporated into Seurat in a future update.

alikhuseynov commented 7 months ago

I was able to get that to work but I'm still getting the issue with the future library

error cannot ffind HDF5 file

Do you have any other suggestions for what could fix it? Greatly appreciate your help so far.

Did it work to make the final object then? And don't use plan("multisession", workers = 10) before loading the object

angie-john commented 7 months ago

Yes I was able to make the object but it was unable to read the hdf5 files still.

alikhuseynov commented 7 months ago

Yes I was able to make the object but it was unable to read the hdf5 files still.

did it read any hdf5 files at all or just didn't read some? it is hard to debug like this and I'm not working on windows machine. Please provide your code or at least final output of function run, and how the object looks like, does it have any FOV etc..? also add sessionInfo(), not just screenshots, one cannot see anything there. and avoid using whitespaces in directory names like mouse brain vizgen data, use _ instead.

angie-john commented 7 months ago

It wasn't able to read any of the hdf5 files. This is my code:

 library(Seurat)
library(ggplot2)
library(data.table)
library(hdf5r)
library(dplyr)
library(BiocParallel)
library(magrittr)
library(progressr)
library(sf)

vizgen.obj <- LoadVizgen(data.dir = "C:/Users/gangliagurdian/Documents/practice/mouse_brain_vizgen_data", 
                         fov = "s2r1", 
                         use.BiocParallel = TRUE,
                         verbose = TRUE,
                         workers.MulticoreParam = 4 ) 
vizgen.obj <- SCTransform(vizgen.obj, assay = "Vizgen", clip.range = c(-10, 10))
vizgen.obj <- RunPCA(vizgen.obj, npcs = 30, features = rownames(vizgen.obj))
vizgen.obj <- RunUMAP(vizgen.obj, dims = 1:30)
vizgen.obj <- FindNeighbors(vizgen.obj, reduction = "pca", dims = 1:30)
vizgen.obj <- FindClusters(vizgen.obj, resolution = 0.3)

DimPlot(vizgen.obj, reduction = "umap")
ImageDimPlot(vizgen.obj, fov = "s2r1", cols = "polychrome", axes = TRUE)

and this was the session info:

R version 4.3.3 (2024-02-29 ucrt)
Platform: x86_64-w64-mingw32/x64 (64-bit)
Running under: Windows 10 x64 (build 19044)

Matrix products: default

locale:
[1] LC_COLLATE=English_United States.utf8  LC_CTYPE=English_United States.utf8    LC_MONETARY=English_United States.utf8
[4] LC_NUMERIC=C                           LC_TIME=English_United States.utf8    

time zone: America/Chicago
tzcode source: internal

attached base packages:
[1] stats     graphics  grDevices utils     datasets  methods   base     

other attached packages:
 [1] SeuratObject_5.0.1  Seurat_4.3.0.9002   sf_1.0-16           progressr_0.14.0    magrittr_2.0.3      BiocParallel_1.36.0
 [7] dplyr_1.1.4         hdf5r_1.3.10        data.table_1.15.4   ggplot2_3.5.0      

loaded via a namespace (and not attached):
  [1] DBI_1.2.2              deldir_2.0-4           pbapply_1.7-2          gridExtra_2.3          rlang_1.1.3           
  [6] RcppAnnoy_0.0.22       spatstat.geom_3.2-9    matrixStats_1.2.0      ggridges_0.5.6         e1071_1.7-14          
 [11] compiler_4.3.3         png_0.1-8              vctrs_0.6.5            reshape2_1.4.4         stringr_1.5.1         
 [16] pkgconfig_2.0.3        fastmap_1.1.1          labeling_0.4.3         utf8_1.2.4             promises_1.2.1        
 [21] tzdb_0.4.0             purrr_1.0.2            bit_4.0.5              jsonlite_1.8.8         goftest_1.2-3         
 [26] later_1.3.2            spatstat.utils_3.0-4   irlba_2.3.5.1          parallel_4.3.3         cluster_2.1.6         
 [31] R6_2.5.1               ica_1.0-3              spatstat.data_3.0-4    stringi_1.8.3          RColorBrewer_1.1-3    
 [36] reticulate_1.35.0      parallelly_1.37.1      lmtest_0.9-40          scattermore_1.2        assertthat_0.2.1      
 [41] Rcpp_1.0.12            tensor_1.5             future.apply_1.11.2    zoo_1.8-12             sctransform_0.4.1     
 [46] httpuv_1.6.15          Matrix_1.6-5           splines_4.3.3          igraph_2.0.3           tidyselect_1.2.1      
 [51] abind_1.4-5            rstudioapi_0.16.0      spatstat.random_3.2-3  spatstat.explore_3.2-7 codetools_0.2-19      
 [56] miniUI_0.1.1.1         listenv_0.9.1          lattice_0.22-5         tibble_3.2.1           plyr_1.8.9            
 [61] shiny_1.8.1.1          withr_3.0.0            ROCR_1.0-11            Rtsne_0.17             future_1.33.2         
 [66] survival_3.5-8         polyclip_1.10-6        units_0.8-5            proxy_0.4-27           fitdistrplus_1.1-11   
 [71] pillar_1.9.0           KernSmooth_2.23-22     plotly_4.10.4          generics_0.1.3         sp_2.1-3              
 [76] munsell_0.5.1          scales_1.3.0           globals_0.16.3         xtable_1.8-4           class_7.3-22          
 [81] glue_1.7.0             lazyeval_0.2.2         tools_4.3.3            RANN_2.6.1             leiden_0.4.3.1        
 [86] dotCall64_1.1-1        cowplot_1.1.3          grid_4.3.3             sfarrow_0.4.1          tidyr_1.3.1           
 [91] tidyverse_2.0.0        colorspace_2.1-0       nlme_3.1-164           patchwork_1.2.0        cli_3.6.2             
 [96] spatstat.sparse_3.0-3  spam_2.10-0            fansi_1.0.6            viridisLite_0.4.2      arrow_15.0.1          
[101] uwot_0.1.16            gtable_0.3.4           digest_0.6.35          classInt_0.4-10        ggrepel_0.9.5         
[106] farver_2.1.1           htmlwidgets_1.6.4      htmltools_0.5.8.1      lifecycle_1.0.4        httr_1.4.7            
[111] mime_0.12              bit64_4.0.5            MASS_7.3-60.0.1
alikhuseynov commented 7 months ago

you have these versions SeuratObject_5.0.1 Seurat_4.3.0.9002

could you show the complete output of messages (not a screenshot) when you run:

vizgen.obj <- LoadVizgen(data.dir = "C:/Users/gangliagurdian/Documents/practice/mouse_brain_vizgen_data", 
                        fov = "s2r1", 
                        use.BiocParallel = TRUE,
                        verbose = TRUE) 

workers.MulticoreParam is not needed since you are on windows machine

check FOVs:

vizgen.obj[[Images(vizgen.obj)[1]]]
vizgen.obj[[Images(vizgen.obj)[1]]][["segmentation"]]
vizgen.obj[[Images(vizgen.obj)[1]]][["centroids"]] %>% str

show the object and plot of a given gene:

vizgen.obj
ImageFeaturePlot(vizgen.obj, features = "gene_name")
angie-john commented 6 months ago

This is the full output after running

vizgen.obj <- LoadVizgen(data.dir = "C:/Users/gangliagurdian/Documents/practice/mouse_brain_vizgen_data", 
                        fov = "s2r1", 
                        use.BiocParallel = TRUE,
                        verbose = TRUE) 
Warning in FUN(...) : Cannot find HDF5 file for field of view 270
Warning in FUN(...) : Cannot find HDF5 file for field of view 271
Warning in FUN(...) : Cannot find HDF5 file for field of view 272
Warning in FUN(...) : Cannot find HDF5 file for field of view 273
Warning in FUN(...) : Cannot find HDF5 file for field of view 274
Warning in FUN(...) : Cannot find HDF5 file for field of view 275
Warning in FUN(...) : Cannot find HDF5 file for field of view 276
Warning in FUN(...) : Cannot find HDF5 file for field of view 277
  |=======================                                                                                                    |  18%Warning in FUN(...) : Cannot find HDF5 file for field of view 278
Warning in FUN(...) : Cannot find HDF5 file for field of view 279
  |=======================                                                                                                    |  19%Warning in FUN(...) : Cannot find HDF5 file for field of view 280
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  |========================                                                                                                   |  19%Warning in FUN(...) : Cannot find HDF5 file for field of view 288
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  |==============================================                                                                             |  37%Warning in FUN(...) : Cannot find HDF5 file for field of view 525
Warning in FUN(...) : Cannot find HDF5 file for field of view 526
Warning in FUN(...) : Cannot find HDF5 file for field of view 527
Warning in FUN(...) : Cannot find HDF5 file for field of view 528
Warning in FUN(...) : Cannot find HDF5 file for field of view 529
Warning in FUN(...) : Cannot find HDF5 file for field of view 530
  |==============================================                                                                             |  38%Warning in FUN(...) : Cannot find HDF5 file for field of view 531
Warning in FUN(...) : Cannot find HDF5 file for field of view 532
Warning in FUN(...) : Cannot find HDF5 file for field of view 533
Warning in FUN(...) : Cannot find HDF5 file for field of view 534
  |===============================================                                                                            |  38%Warning in FUN(...) : Cannot find HDF5 file for field of view 535
Warning in FUN(...) : Cannot find HDF5 file for field of view 536
Warning in FUN(...) : Cannot find HDF5 file for field of view 537
Warning in FUN(...) : Cannot find HDF5 file for field of view 538
Warning in FUN(...) : Cannot find HDF5 file for field of view 539
Warning in FUN(...) : Cannot find HDF5 file for field of view 540
Warning in FUN(...) : Cannot find HDF5 file for field of view 541
Warning in FUN(...) : Cannot find HDF5 file for field of view 542
Warning in FUN(...) : Cannot find HDF5 file for field of view 543
  |===============================================                                                                            |  39%Warning in FUN(...) : Cannot find HDF5 file for field of view 544
  |================================================                                                                           |  39%Warning in FUN(...) : Cannot find HDF5 file for field of view 545
Warning in FUN(...) : Cannot find HDF5 file for field of view 546
Warning in FUN(...) : Cannot find HDF5 file for field of view 547
Warning in FUN(...) : Cannot find HDF5 file for field of view 548
Warning in FUN(...) : Cannot find HDF5 file for field of view 549
Warning in FUN(...) : Cannot find HDF5 file for field of view 550
Warning in FUN(...) : Cannot find HDF5 file for field of view 551
Warning in FUN(...) : Cannot find HDF5 file for field of view 552
Warning in FUN(...) : Cannot find HDF5 file for field of view 553
Warning in FUN(...) : Cannot find HDF5 file for field of view 554
  |=================================================                                                                          |  39%Warning in FUN(...) : Cannot find HDF5 file for field of view 555
  |=================================================                                                                          |  40%Warning in FUN(...) : Cannot find HDF5 file for field of view 556
Warning in FUN(...) : Cannot find HDF5 file for field of view 557
Warning in FUN(...) : Cannot find HDF5 file for field of view 558
Warning in FUN(...) : Cannot find HDF5 file for field of view 559
Warning in FUN(...) : Cannot find HDF5 file for field of view 560
Warning in FUN(...) : Cannot find HDF5 file for field of view 561
Warning in FUN(...) : Cannot find HDF5 file for field of view 562
Warning in FUN(...) : Cannot find HDF5 file for field of view 563
Warning in FUN(...) : Cannot find HDF5 file for field of view 564
  |==================================================                                                                         |  40%Warning in FUN(...) : Cannot find HDF5 file for field of view 565
Warning in FUN(...) : Cannot find HDF5 file for field of view 566
Warning in FUN(...) : Cannot find HDF5 file for field of view 567
Warning in FUN(...) : Cannot find HDF5 file for field of view 568
  |==================================================                                                                         |  41%Warning in FUN(...) : Cannot find HDF5 file for field of view 569
Warning in FUN(...) : Cannot find HDF5 file for field of view 570
Warning in FUN(...) : Cannot find HDF5 file for field of view 571
Warning in FUN(...) : Cannot find HDF5 file for field of view 572
Warning in FUN(...) : Cannot find HDF5 file for field of view 577
Warning in FUN(...) : Cannot find HDF5 file for field of view 578
  |===================================================                                                                        |  41%Warning in FUN(...) : Cannot find HDF5 file for field of view 579
Warning in FUN(...) : Cannot find HDF5 file for field of view 580
Warning in FUN(...) : Cannot find HDF5 file for field of view 581
Warning in FUN(...) : Cannot find HDF5 file for field of view 582
Warning in FUN(...) : Cannot find HDF5 file for field of view 583
Warning in FUN(...) : Cannot find HDF5 file for field of view 584
  |===================================================                                                                        |  42%Warning in FUN(...) : Cannot find HDF5 file for field of view 585
Warning in FUN(...) : Cannot find HDF5 file for field of view 586
Warning in FUN(...) : Cannot find HDF5 file for field of view 587
Warning in FUN(...) : Cannot find HDF5 file for field of view 588
Warning in FUN(...) : Cannot find HDF5 file for field of view 589
  |====================================================                                                                       |  42%Warning in FUN(...) : Cannot find HDF5 file for field of view 590
Warning in FUN(...) : Cannot find HDF5 file for field of view 591
Warning in FUN(...) : Cannot find HDF5 file for field of view 592
Warning in FUN(...) : Cannot find HDF5 file for field of view 593
Warning in FUN(...) : Cannot find HDF5 file for field of view 594
Warning in FUN(...) : Cannot find HDF5 file for field of view 595
Warning in FUN(...) : Cannot find HDF5 file for field of view 596
  |====================================================                                                                       |  43%Warning in FUN(...) : Cannot find HDF5 file for field of view 597
Warning in FUN(...) : Cannot find HDF5 file for field of view 598
Warning in FUN(...) : Cannot find HDF5 file for field of view 599
  |=====================================================                                                                      |  43%Warning in FUN(...) : Cannot find HDF5 file for field of view 600
Warning in FUN(...) : Cannot find HDF5 file for field of view 601
Warning in FUN(...) : Cannot find HDF5 file for field of view 602
Warning in FUN(...) : Cannot find HDF5 file for field of view 603
Warning in FUN(...) : Cannot find HDF5 file for field of view 604
Warning in FUN(...) : Cannot find HDF5 file for field of view 605
Warning in FUN(...) : Cannot find HDF5 file for field of view 606
Warning in FUN(...) : Cannot find HDF5 file for field of view 607
Warning in FUN(...) : Cannot find HDF5 file for field of view 608
Warning in FUN(...) : Cannot find HDF5 file for field of view 609
  |======================================================                                                                     |  44%Warning in FUN(...) : Cannot find HDF5 file for field of view 610
Warning in FUN(...) : Cannot find HDF5 file for field of view 611
Warning in FUN(...) : Cannot find HDF5 file for field of view 612
Warning in FUN(...) : Cannot find HDF5 file for field of view 613
Warning in FUN(...) : Cannot find HDF5 file for field of view 614
Warning in FUN(...) : Cannot find HDF5 file for field of view 615
Warning in FUN(...) : Cannot find HDF5 file for field of view 616
Warning in FUN(...) : Cannot find HDF5 file for field of view 617
Warning in FUN(...) : Cannot find HDF5 file for field of view 618
Warning in FUN(...) : Cannot find HDF5 file for field of view 619
  |=======================================================                                                                    |  44%Warning in FUN(...) : Cannot find HDF5 file for field of view 620
Warning in FUN(...) : Cannot find HDF5 file for field of view 621
  |=======================================================                                                                    |  45%Warning in FUN(...) : Cannot find HDF5 file for field of view 622
Warning in FUN(...) : Cannot find HDF5 file for field of view 623
Warning in FUN(...) : Cannot find HDF5 file for field of view 624
Warning in FUN(...) : Cannot find HDF5 file for field of view 625
Warning in FUN(...) : Cannot find HDF5 file for field of view 626
Warning in FUN(...) : Cannot find HDF5 file for field of view 627
Warning in FUN(...) : Cannot find HDF5 file for field of view 628
Warning in FUN(...) : Cannot find HDF5 file for field of view 629
  |========================================================                                                                   |  45%Warning in FUN(...) : Cannot find HDF5 file for field of view 630
Warning in FUN(...) : Cannot find HDF5 file for field of view 631
Warning in FUN(...) : Cannot find HDF5 file for field of view 632
Warning in FUN(...) : Cannot find HDF5 file for field of view 633
Warning in FUN(...) : Cannot find HDF5 file for field of view 634
  |========================================================                                                                   |  46%Warning in FUN(...) : Cannot find HDF5 file for field of view 635
Warning in FUN(...) : Cannot find HDF5 file for field of view 636
Warning in FUN(...) : Cannot find HDF5 file for field of view 637
Warning in FUN(...) : Cannot find HDF5 file for field of view 638
Warning in FUN(...) : Cannot find HDF5 file for field of view 639
  |=========================================================                                                                  |  46%Warning in FUN(...) : Cannot find HDF5 file for field of view 640
Warning in FUN(...) : Cannot find HDF5 file for field of view 641
Warning in FUN(...) : Cannot find HDF5 file for field of view 642
Warning in FUN(...) : Cannot find HDF5 file for field of view 647
Warning in FUN(...) : Cannot find HDF5 file for field of view 648
Warning in FUN(...) : Cannot find HDF5 file for field of view 649
Warning in FUN(...) : Cannot find HDF5 file for field of view 650
  |=========================================================                                                                  |  47%Warning in FUN(...) : Cannot find HDF5 file for field of view 651
Warning in FUN(...) : Cannot find HDF5 file for field of view 652
Warning in FUN(...) : Cannot find HDF5 file for field of view 653
  |==========================================================                                                                 |  47%Warning in FUN(...) : Cannot find HDF5 file for field of view 654
Warning in FUN(...) : Cannot find HDF5 file for field of view 655
Warning in FUN(...) : Cannot find HDF5 file for field of view 656
Warning in FUN(...) : Cannot find HDF5 file for field of view 657
Warning in FUN(...) : Cannot find HDF5 file for field of view 658
Warning in FUN(...) : Cannot find HDF5 file for field of view 659
Warning in FUN(...) : Cannot find HDF5 file for field of view 660
Warning in FUN(...) : Cannot find HDF5 file for field of view 661
Warning in FUN(...) : Cannot find HDF5 file for field of view 662
  |==========================================================                                                                 |  48%Warning in FUN(...) : Cannot find HDF5 file for field of view 663
  |===========================================================                                                                |  48%Warning in FUN(...) : Cannot find HDF5 file for field of view 664
Warning in FUN(...) : Cannot find HDF5 file for field of view 665
Warning in FUN(...) : Cannot find HDF5 file for field of view 666
Warning in FUN(...) : Cannot find HDF5 file for field of view 667
Warning in FUN(...) : Cannot find HDF5 file for field of view 668
Warning in FUN(...) : Cannot find HDF5 file for field of view 669
Warning in FUN(...) : Cannot find HDF5 file for field of view 670
Warning in FUN(...) : Cannot find HDF5 file for field of view 671
Warning in FUN(...) : Cannot find HDF5 file for field of view 672
Warning in FUN(...) : Cannot find HDF5 file for field of view 673
  |============================================================                                                               |  48%Warning in FUN(...) : Cannot find HDF5 file for field of view 674
Warning in FUN(...) : Cannot find HDF5 file for field of view 675
  |============================================================                                                               |  49%Warning in FUN(...) : Cannot find HDF5 file for field of view 676
Warning in FUN(...) : Cannot find HDF5 file for field of view 677
Warning in FUN(...) : Cannot find HDF5 file for field of view 678
Warning in FUN(...) : Cannot find HDF5 file for field of view 679
Warning in FUN(...) : Cannot find HDF5 file for field of view 680
Warning in FUN(...) : Cannot find HDF5 file for field of view 681
Warning in FUN(...) : Cannot find HDF5 file for field of view 682
Warning in FUN(...) : Cannot find HDF5 file for field of view 683
  |=============================================================                                                              |  49%Warning in FUN(...) : Cannot find HDF5 file for field of view 684
Warning in FUN(...) : Cannot find HDF5 file for field of view 685
Warning in FUN(...) : Cannot find HDF5 file for field of view 686
Warning in FUN(...) : Cannot find HDF5 file for field of view 687
  |=============================================================                                                              |  50%Warning in FUN(...) : Cannot find HDF5 file for field of view 688
Warning in FUN(...) : Cannot find HDF5 file for field of view 689
Warning in FUN(...) : Cannot find HDF5 file for field of view 690
Warning in FUN(...) : Cannot find HDF5 file for field of view 691
Warning in FUN(...) : Cannot find HDF5 file for field of view 692
Warning in FUN(...) : Cannot find HDF5 file for field of view 693
  |==============================================================                                                             |  50%Warning in FUN(...) : Cannot find HDF5 file for field of view 694
Warning in FUN(...) : Cannot find HDF5 file for field of view 695
Warning in FUN(...) : Cannot find HDF5 file for field of view 696
Warning in FUN(...) : Cannot find HDF5 file for field of view 697
Warning in FUN(...) : Cannot find HDF5 file for field of view 698
Warning in FUN(...) : Cannot find HDF5 file for field of view 699
Warning in FUN(...) : Cannot find HDF5 file for field of view 700
  |==============================================================                                                             |  51%Warning in FUN(...) : Cannot find HDF5 file for field of view 701
Warning in FUN(...) : Cannot find HDF5 file for field of view 702
Warning in FUN(...) : Cannot find HDF5 file for field of view 703
Warning in FUN(...) : Cannot find HDF5 file for field of view 704
  |===============================================================                                                            |  51%Warning in FUN(...) : Cannot find HDF5 file for field of view 705
Warning in FUN(...) : Cannot find HDF5 file for field of view 706
Warning in FUN(...) : Cannot find HDF5 file for field of view 707
Warning in FUN(...) : Cannot find HDF5 file for field of view 708
Warning in FUN(...) : Cannot find HDF5 file for field of view 709
Warning in FUN(...) : Cannot find HDF5 file for field of view 710
Warning in FUN(...) : Cannot find HDF5 file for field of view 711
Warning in FUN(...) : Cannot find HDF5 file for field of view 712
  |===============================================================                                                            |  52%Warning in FUN(...) : Cannot find HDF5 file for field of view 716
Warning in FUN(...) : Cannot find HDF5 file for field of view 717
  |================================================================                                                           |  52%Warning in FUN(...) : Cannot find HDF5 file for field of view 718
Warning in FUN(...) : Cannot find HDF5 file for field of view 719
Warning in FUN(...) : Cannot find HDF5 file for field of view 720
Warning in FUN(...) : Cannot find HDF5 file for field of view 721
Warning in FUN(...) : Cannot find HDF5 file for field of view 722
Warning in FUN(...) : Cannot find HDF5 file for field of view 723
Warning in FUN(...) : Cannot find HDF5 file for field of view 724
Warning in FUN(...) : Cannot find HDF5 file for field of view 725
Warning in FUN(...) : Cannot find HDF5 file for field of view 726
Warning in FUN(...) : Cannot find HDF5 file for field of view 727
  |=================================================================                                                          |  52%Warning in FUN(...) : Cannot find HDF5 file for field of view 728
  |=================================================================                                                          |  53%Warning in FUN(...) : Cannot find HDF5 file for field of view 729
Warning in FUN(...) : Cannot find HDF5 file for field of view 730
Warning in FUN(...) : Cannot find HDF5 file for field of view 731
Warning in FUN(...) : Cannot find HDF5 file for field of view 732
Warning in FUN(...) : Cannot find HDF5 file for field of view 733
Warning in FUN(...) : Cannot find HDF5 file for field of view 734
Warning in FUN(...) : Cannot find HDF5 file for field of view 735
Warning in FUN(...) : Cannot find HDF5 file for field of view 736
Warning in FUN(...) : Cannot find HDF5 file for field of view 737
  |==================================================================                                                         |  53%Warning in FUN(...) : Cannot find HDF5 file for field of view 738
Warning in FUN(...) : Cannot find HDF5 file for field of view 739
Warning in FUN(...) : Cannot find HDF5 file for field of view 740
  |==================================================================                                                         |  54%Warning in FUN(...) : Cannot find HDF5 file for field of view 741
Warning in FUN(...) : Cannot find HDF5 file for field of view 742
Warning in FUN(...) : Cannot find HDF5 file for field of view 743
Warning in FUN(...) : Cannot find HDF5 file for field of view 744
Warning in FUN(...) : Cannot find HDF5 file for field of view 745
Warning in FUN(...) : Cannot find HDF5 file for field of view 746
Warning in FUN(...) : Cannot find HDF5 file for field of view 747
  |===================================================================                                                        |  54%Warning in FUN(...) : Cannot find HDF5 file for field of view 748
Warning in FUN(...) : Cannot find HDF5 file for field of view 749
Warning in FUN(...) : Cannot find HDF5 file for field of view 750
Warning in FUN(...) : Cannot find HDF5 file for field of view 751
Warning in FUN(...) : Cannot find HDF5 file for field of view 752
  |===================================================================                                                        |  55%Warning in FUN(...) : Cannot find HDF5 file for field of view 753
Warning in FUN(...) : Cannot find HDF5 file for field of view 754
Warning in FUN(...) : Cannot find HDF5 file for field of view 755
Warning in FUN(...) : Cannot find HDF5 file for field of view 756
Warning in FUN(...) : Cannot find HDF5 file for field of view 757
  |====================================================================                                                       |  55%Warning in FUN(...) : Cannot find HDF5 file for field of view 758
Warning in FUN(...) : Cannot find HDF5 file for field of view 759
Warning in FUN(...) : Cannot find HDF5 file for field of view 760
Warning in FUN(...) : Cannot find HDF5 file for field of view 761
Warning in FUN(...) : Cannot find HDF5 file for field of view 762
Warning in FUN(...) : Cannot find HDF5 file for field of view 763
Warning in FUN(...) : Cannot find HDF5 file for field of view 764
Warning in FUN(...) : Cannot find HDF5 file for field of view 765
  |====================================================================                                                       |  56%Warning in FUN(...) : Cannot find HDF5 file for field of view 766
Warning in FUN(...) : Cannot find HDF5 file for field of view 767
  |=====================================================================                                                      |  56%Warning in FUN(...) : Cannot find HDF5 file for field of view 768
Warning in FUN(...) : Cannot find HDF5 file for field of view 769
Warning in FUN(...) : Cannot find HDF5 file for field of view 770
Warning in FUN(...) : Cannot find HDF5 file for field of view 771
Warning in FUN(...) : Cannot find HDF5 file for field of view 772
Warning in FUN(...) : Cannot find HDF5 file for field of view 773
Warning in FUN(...) : Cannot find HDF5 file for field of view 774
Warning in FUN(...) : Cannot find HDF5 file for field of view 775
Warning in FUN(...) : Cannot find HDF5 file for field of view 776
Warning in FUN(...) : Cannot find HDF5 file for field of view 777
  |======================================================================                                                     |  57%Warning in FUN(...) : Cannot find HDF5 file for field of view 778
Warning in FUN(...) : Cannot find HDF5 file for field of view 779
Warning in FUN(...) : Cannot find HDF5 file for field of view 780
Warning in FUN(...) : Cannot find HDF5 file for field of view 781
Warning in FUN(...) : Cannot find HDF5 file for field of view 785
Warning in FUN(...) : Cannot find HDF5 file for field of view 786
Warning in FUN(...) : Cannot find HDF5 file for field of view 787
Warning in FUN(...) : Cannot find HDF5 file for field of view 788
Warning in FUN(...) : Cannot find HDF5 file for field of view 789
Warning in FUN(...) : Cannot find HDF5 file for field of view 790
  |=======================================================================                                                    |  57%Warning in FUN(...) : Cannot find HDF5 file for field of view 791
Warning in FUN(...) : Cannot find HDF5 file for field of view 792
Warning in FUN(...) : Cannot find HDF5 file for field of view 793
  |=======================================================================                                                    |  58%Warning in FUN(...) : Cannot find HDF5 file for field of view 794
Warning in FUN(...) : Cannot find HDF5 file for field of view 795
Warning in FUN(...) : Cannot find HDF5 file for field of view 796
Warning in FUN(...) : Cannot find HDF5 file for field of view 797
Warning in FUN(...) : Cannot find HDF5 file for field of view 798
Warning in FUN(...) : Cannot find HDF5 file for field of view 799
Warning in FUN(...) : Cannot find HDF5 file for field of view 800
  |========================================================================                                                   |  58%Warning in FUN(...) : Cannot find HDF5 file for field of view 801
Warning in FUN(...) : Cannot find HDF5 file for field of view 802
Warning in FUN(...) : Cannot find HDF5 file for field of view 803
Warning in FUN(...) : Cannot find HDF5 file for field of view 804
Warning in FUN(...) : Cannot find HDF5 file for field of view 805
  |========================================================================                                                   |  59%Warning in FUN(...) : Cannot find HDF5 file for field of view 806
Warning in FUN(...) : Cannot find HDF5 file for field of view 807
Warning in FUN(...) : Cannot find HDF5 file for field of view 808
Warning in FUN(...) : Cannot find HDF5 file for field of view 809
Warning in FUN(...) : Cannot find HDF5 file for field of view 810
Warning in FUN(...) : Cannot find HDF5 file for field of view 811
  |=========================================================================                                                  |  59%Warning in FUN(...) : Cannot find HDF5 file for field of view 812
Warning in FUN(...) : Cannot find HDF5 file for field of view 813
Warning in FUN(...) : Cannot find HDF5 file for field of view 814
Warning in FUN(...) : Cannot find HDF5 file for field of view 815
Warning in FUN(...) : Cannot find HDF5 file for field of view 816
Warning in FUN(...) : Cannot find HDF5 file for field of view 819
  |=========================================================================                                                  |  60%Warning in FUN(...) : Cannot find HDF5 file for field of view 820
Warning in FUN(...) : Cannot find HDF5 file for field of view 821
Warning in FUN(...) : Cannot find HDF5 file for field of view 822
Warning in FUN(...) : Cannot find HDF5 file for field of view 823
  |==========================================================================                                                 |  60%Warning in FUN(...) : Cannot find HDF5 file for field of view 824
Warning in FUN(...) : Cannot find HDF5 file for field of view 825
Warning in FUN(...) : Cannot find HDF5 file for field of view 826
Warning in FUN(...) : Cannot find HDF5 file for field of view 827
Warning in FUN(...) : Cannot find HDF5 file for field of view 828
Warning in FUN(...) : Cannot find HDF5 file for field of view 829
Warning in FUN(...) : Cannot find HDF5 file for field of view 830
Warning in FUN(...) : Cannot find HDF5 file for field of view 831
Warning in FUN(...) : Cannot find HDF5 file for field of view 832
  |==========================================================================                                                 |  61%Warning in FUN(...) : Cannot find HDF5 file for field of view 833
  |===========================================================================                                                |  61%Warning in FUN(...) : Cannot find HDF5 file for field of view 834
Warning in FUN(...) : Cannot find HDF5 file for field of view 835
Warning in FUN(...) : Cannot find HDF5 file for field of view 836
Warning in FUN(...) : Cannot find HDF5 file for field of view 837
Warning in FUN(...) : Cannot find HDF5 file for field of view 838
Warning in FUN(...) : Cannot find HDF5 file for field of view 839
Warning in FUN(...) : Cannot find HDF5 file for field of view 840
Warning in FUN(...) : Cannot find HDF5 file for field of view 841
Warning in FUN(...) : Cannot find HDF5 file for field of view 842
Warning in FUN(...) : Cannot find HDF5 file for field of view 843
  |============================================================================                                               |  61%Warning in FUN(...) : Cannot find HDF5 file for field of view 844
  |============================================================================                                               |  62%Warning in FUN(...) : Cannot find HDF5 file for field of view 845
Warning in FUN(...) : Cannot find HDF5 file for field of view 846
Warning in FUN(...) : Cannot find HDF5 file for field of view 847
Warning in FUN(...) : Cannot find HDF5 file for field of view 848
Warning in FUN(...) : Cannot find HDF5 file for field of view 849
Warning in FUN(...) : Cannot find HDF5 file for field of view 850
Warning in FUN(...) : Cannot find HDF5 file for field of view 855
Warning in FUN(...) : Cannot find HDF5 file for field of view 856
Warning in FUN(...) : Cannot find HDF5 file for field of view 857
  |=============================================================================                                              |  62%Warning in FUN(...) : Cannot find HDF5 file for field of view 858
Warning in FUN(...) : Cannot find HDF5 file for field of view 859
Warning in FUN(...) : Cannot find HDF5 file for field of view 860
Warning in FUN(...) : Cannot find HDF5 file for field of view 861
  |=============================================================================                                              |  63%Warning in FUN(...) : Cannot find HDF5 file for field of view 862
Warning in FUN(...) : Cannot find HDF5 file for field of view 863
Warning in FUN(...) : Cannot find HDF5 file for field of view 864
Warning in FUN(...) : Cannot find HDF5 file for field of view 865
Warning in FUN(...) : Cannot find HDF5 file for field of view 866
Warning in FUN(...) : Cannot find HDF5 file for field of view 867
  |==============================================================================                                             |  63%Warning in FUN(...) : Cannot find HDF5 file for field of view 868
Warning in FUN(...) : Cannot find HDF5 file for field of view 869
Warning in FUN(...) : Cannot find HDF5 file for field of view 870
Warning in FUN(...) : Cannot find HDF5 file for field of view 871
Warning in FUN(...) : Cannot find HDF5 file for field of view 872
Warning in FUN(...) : Cannot find HDF5 file for field of view 873
  |==============================================================================                                             |  64%Warning in FUN(...) : Cannot find HDF5 file for field of view 874
Warning in FUN(...) : Cannot find HDF5 file for field of view 875
Warning in FUN(...) : Cannot find HDF5 file for field of view 876
Warning in FUN(...) : Cannot find HDF5 file for field of view 877
  |===============================================================================                                            |  64%Warning in FUN(...) : Cannot find HDF5 file for field of view 878
Warning in FUN(...) : Cannot find HDF5 file for field of view 879
Warning in FUN(...) : Cannot find HDF5 file for field of view 880
Warning in FUN(...) : Cannot find HDF5 file for field of view 881
Warning in FUN(...) : Cannot find HDF5 file for field of view 882
Warning in FUN(...) : Cannot find HDF5 file for field of view 883
Warning in FUN(...) : Cannot find HDF5 file for field of view 884
Warning in FUN(...) : Cannot find HDF5 file for field of view 885
Warning in FUN(...) : Cannot find HDF5 file for field of view 886
  |===============================================================================                                            |  65%Warning in FUN(...) : Cannot find HDF5 file for field of view 887
  |================================================================================                                           |  65%Warning in FUN(...) : Cannot find HDF5 file for field of view 888
Warning in FUN(...) : Cannot find HDF5 file for field of view 889
Warning in FUN(...) : Cannot find HDF5 file for field of view 890
Warning in FUN(...) : Cannot find HDF5 file for field of view 891
Warning in FUN(...) : Cannot find HDF5 file for field of view 892
Warning in FUN(...) : Cannot find HDF5 file for field of view 893
Warning in FUN(...) : Cannot find HDF5 file for field of view 894
Warning in FUN(...) : Cannot find HDF5 file for field of view 895
Warning in FUN(...) : Cannot find HDF5 file for field of view 896
Warning in FUN(...) : Cannot find HDF5 file for field of view 897
  |=================================================================================                                          |  65%Warning in FUN(...) : Cannot find HDF5 file for field of view 898
  |=================================================================================                                          |  66%Warning in FUN(...) : Cannot find HDF5 file for field of view 899
Warning in FUN(...) : Cannot find HDF5 file for field of view 900
Warning in FUN(...) : Cannot find HDF5 file for field of view 901
Warning in FUN(...) : Cannot find HDF5 file for field of view 902
Warning in FUN(...) : Cannot find HDF5 file for field of view 903
Warning in FUN(...) : Cannot find HDF5 file for field of view 904
Warning in FUN(...) : Cannot find HDF5 file for field of view 905
Warning in FUN(...) : Cannot find HDF5 file for field of view 906
Warning in FUN(...) : Cannot find HDF5 file for field of view 907
  |==================================================================================                                         |  66%Warning in FUN(...) : Cannot find HDF5 file for field of view 908
Warning in FUN(...) : Cannot find HDF5 file for field of view 909
Warning in FUN(...) : Cannot find HDF5 file for field of view 910
  |==================================================================================                                         |  67%Warning in FUN(...) : Cannot find HDF5 file for field of view 911
Warning in FUN(...) : Cannot find HDF5 file for field of view 912
Warning in FUN(...) : Cannot find HDF5 file for field of view 913
Warning in FUN(...) : Cannot find HDF5 file for field of view 914
Warning in FUN(...) : Cannot find HDF5 file for field of view 915
Warning in FUN(...) : Cannot find HDF5 file for field of view 916
Warning in FUN(...) : Cannot find HDF5 file for field of view 917
Warning in FUN(...) : Cannot find HDF5 file for field of view 918
  |===================================================================================                                        |  67%Warning in FUN(...) : Cannot find HDF5 file for field of view 919
Warning in FUN(...) : Cannot find HDF5 file for field of view 920
Warning in FUN(...) : Cannot find HDF5 file for field of view 921
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  |===================================================================================                                        |  68%Warning in FUN(...) : Cannot find HDF5 file for field of view 927
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  |====================================================================================                                       |  68%Warning in FUN(...) : Cannot find HDF5 file for field of view 932
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  |====================================================================================                                       |  69%Warning in FUN(...) : Cannot find HDF5 file for field of view 939
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  |=====================================================================================                                      |  69%Warning in FUN(...) : Cannot find HDF5 file for field of view 942
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  |======================================================================================                                     |  70%Warning in FUN(...) : Cannot find HDF5 file for field of view 952
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  |=======================================================================================                                    |  70%Warning in FUN(...) : Cannot find HDF5 file for field of view 962
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  |=======================================================================================                                    |  71%Warning in FUN(...) : Cannot find HDF5 file for field of view 964
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  |========================================================================================                                   |  71%Warning in FUN(...) : Cannot find HDF5 file for field of view 972
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  |========================================================================================                                   |  72%Warning in FUN(...) : Cannot find HDF5 file for field of view 977
Warning in FUN(...) : Cannot find HDF5 file for field of view 978
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  |=========================================================================================                                  |  72%Warning in FUN(...) : Cannot find HDF5 file for field of view 982
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  |=========================================================================================                                  |  73%Warning in FUN(...) : Cannot find HDF5 file for field of view 989
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  |==========================================================================================                                 |  73%Warning in FUN(...) : Cannot find HDF5 file for field of view 994
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  |===============================================================================================                            |  77%Warning in FUN(...) : Cannot find HDF5 file for field of view 1045
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  |===============================================================================================                            |  78%Warning in FUN(...) : Cannot find HDF5 file for field of view 1053
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  |================================================================================================                           |  78%Warning in FUN(...) : Cannot find HDF5 file for field of view 1055
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  |=================================================================================================                          |  79%Warning in FUN(...) : Cannot find HDF5 file for field of view 1069
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  |==================================================================================================                         |  79%Warning in FUN(...) : Cannot find HDF5 file for field of view 1079
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  |==================================================================================================                         |  80%Warning in FUN(...) : Cannot find HDF5 file for field of view 1082
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  |===================================================================================================                        |  80%Warning in FUN(...) : Cannot find HDF5 file for field of view 1089
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  |===================================================================================================                        |  81%Warning in FUN(...) : Cannot find HDF5 file for field of view 1094
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  |====================================================================================================                       |  82%Warning in FUN(...) : Cannot find HDF5 file for field of view 1107
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  |======================================================================================================                     |  83%Warning in FUN(...) : Cannot find HDF5 file for field of view 1119
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  |=======================================================================================================                    |  83%Warning in FUN(...) : Cannot find HDF5 file for field of view 1133
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  |========================================================================================================                   |  84%Warning in FUN(...) : Cannot find HDF5 file for field of view 1143
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  |========================================================================================================                   |  85%Warning in FUN(...) : Cannot find HDF5 file for field of view 1147
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  |=========================================================================================================                  |  85%Warning in FUN(...) : Cannot find HDF5 file for field of view 1153
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  |=========================================================================================================                  |  86%Warning in FUN(...) : Cannot find HDF5 file for field of view 1159
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  |==========================================================================================================                 |  86%Warning in FUN(...) : Cannot find HDF5 file for field of view 1163
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  |==========================================================================================================                 |  87%Warning in FUN(...) : Cannot find HDF5 file for field of view 1172
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  |============================================================================================================               |  87%Warning in FUN(...) : Cannot find HDF5 file for field of view 1183
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  |=============================================================================================================              |  88%Warning in FUN(...) : Cannot find HDF5 file for field of view 1196
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  |=============================================================================================================              |  89%Warning in FUN(...) : Cannot find HDF5 file for field of view 1200
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  |==============================================================================================================             |  89%Warning in FUN(...) : Cannot find HDF5 file for field of view 1206
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  |==============================================================================================================             |  90%Warning in FUN(...) : Cannot find HDF5 file for field of view 1212
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  |===============================================================================================================            |  90%Warning in FUN(...) : Cannot find HDF5 file for field of view 1216
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  |===============================================================================================================            |  91%Warning in FUN(...) : Cannot find HDF5 file for field of view 1226
  |================================================================================================================           |  91%Warning in FUN(...) : Cannot find HDF5 file for field of view 1227
Warning in FUN(...) : Cannot find HDF5 file for field of view 1228
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Warning in FUN(...) : Cannot find HDF5 file for field of view 1236
  |=================================================================================================================          |  91%Warning in FUN(...) : Cannot find HDF5 file for field of view 1237
  |=================================================================================================================          |  92%Warning in FUN(...) : Cannot find HDF5 file for field of view 1238
Warning in FUN(...) : Cannot find HDF5 file for field of view 1239
Warning in FUN(...) : Cannot find HDF5 file for field of view 1240
Warning in FUN(...) : Cannot find HDF5 file for field of view 1241
Warning in FUN(...) : Cannot find HDF5 file for field of view 1242
Warning in FUN(...) : Cannot find HDF5 file for field of view 1243
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Warning in FUN(...) : Cannot find HDF5 file for field of view 1245
Warning in FUN(...) : Cannot find HDF5 file for field of view 1250
Warning in FUN(...) : Cannot find HDF5 file for field of view 1251
  |==================================================================================================================         |  92%Warning in FUN(...) : Cannot find HDF5 file for field of view 1252
Warning in FUN(...) : Cannot find HDF5 file for field of view 1253
  |==================================================================================================================         |  93%Warning in FUN(...) : Cannot find HDF5 file for field of view 1254
Warning in FUN(...) : Cannot find HDF5 file for field of view 1255
Warning in FUN(...) : Cannot find HDF5 file for field of view 1256
Warning in FUN(...) : Cannot find HDF5 file for field of view 1257
Warning in FUN(...) : Cannot find HDF5 file for field of view 1258
Warning in FUN(...) : Cannot find HDF5 file for field of view 1259
Warning in FUN(...) : Cannot find HDF5 file for field of view 1260
Warning in FUN(...) : Cannot find HDF5 file for field of view 1261
  |===================================================================================================================        |  93%Warning in FUN(...) : Cannot find HDF5 file for field of view 1262
Warning in FUN(...) : Cannot find HDF5 file for field of view 1263
Warning in FUN(...) : Cannot find HDF5 file for field of view 1264
Warning in FUN(...) : Cannot find HDF5 file for field of view 1265
Warning in FUN(...) : Cannot find HDF5 file for field of view 1266
  |===================================================================================================================        |  94%Warning in FUN(...) : Cannot find HDF5 file for field of view 1267
Warning in FUN(...) : Cannot find HDF5 file for field of view 1268
Warning in FUN(...) : Cannot find HDF5 file for field of view 1269
Warning in FUN(...) : Cannot find HDF5 file for field of view 1270
Warning in FUN(...) : Cannot find HDF5 file for field of view 1271
  |====================================================================================================================       |  94%Warning in FUN(...) : Cannot find HDF5 file for field of view 1272
Warning in FUN(...) : Cannot find HDF5 file for field of view 1275
Warning in FUN(...) : Cannot find HDF5 file for field of view 1276
Warning in FUN(...) : Cannot find HDF5 file for field of view 1277
Warning in FUN(...) : Cannot find HDF5 file for field of view 1278
Warning in FUN(...) : Cannot find HDF5 file for field of view 1279
Warning in FUN(...) : Cannot find HDF5 file for field of view 1280
  |====================================================================================================================       |  95%Warning in FUN(...) : Cannot find HDF5 file for field of view 1281
Warning in FUN(...) : Cannot find HDF5 file for field of view 1282
Warning in FUN(...) : Cannot find HDF5 file for field of view 1283
  |=====================================================================================================================      |  95%Warning in FUN(...) : Cannot find HDF5 file for field of view 1284
Warning in FUN(...) : Cannot find HDF5 file for field of view 1285
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Warning in FUN(...) : Cannot find HDF5 file for field of view 1287
Warning in FUN(...) : Cannot find HDF5 file for field of view 1288
Warning in FUN(...) : Cannot find HDF5 file for field of view 1289
Warning in FUN(...) : Cannot find HDF5 file for field of view 1290
Warning in FUN(...) : Cannot find HDF5 file for field of view 1291
Warning in FUN(...) : Cannot find HDF5 file for field of view 1292
Warning in FUN(...) : Cannot find HDF5 file for field of view 1293
  |======================================================================================================================     |  96%Warning in FUN(...) : Cannot find HDF5 file for field of view 1294
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Warning in FUN(...) : Cannot find HDF5 file for field of view 1302
Warning in FUN(...) : Cannot find HDF5 file for field of view 1303
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  |=======================================================================================================================    |  96%Warning in FUN(...) : Cannot find HDF5 file for field of view 1309
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  |=======================================================================================================================    |  97%Warning in FUN(...) : Cannot find HDF5 file for field of view 1311
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  |========================================================================================================================   |  97%Warning in FUN(...) : Cannot find HDF5 file for field of view 1319
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  |========================================================================================================================   |  98%Warning in FUN(...) : Cannot find HDF5 file for field of view 1324
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  |=========================================================================================================================  |  98%Warning in FUN(...) : Cannot find HDF5 file for field of view 1330
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  |=========================================================================================================================  |  99%Warning in FUN(...) : Cannot find HDF5 file for field of view 1337
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  |========================================================================================================================== |  99%Warning in FUN(...) : Cannot find HDF5 file for field of view 1346
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Warning in FUN(...) : Cannot find HDF5 file for field of view 1354
  |========================================================================================================================== | 100%Warning in FUN(...) : Cannot find HDF5 file for field of view 1355
  |===========================================================================================================================| 100%Warning in FUN(...) : Cannot find HDF5 file for field of view 1359
Warning in FUN(...) : Cannot find HDF5 file for field of view 1360
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Warning in ReadVizgen(data.dir = data.dir, mol.type = mol.type, filter = filter,  :
  80223 cells missing polygon information
Creating Seurat object..
Creating FOVs..

>>> using segmentations
>>> adding metadata infos
>>> adding FOV
>>> all counts are > 0
Updating object:
Validating object structure
Updating object slots
Ensuring keys are in the proper structure
Ensuring keys are in the proper structure
Ensuring feature names don't have underscores or pipes
Updating slots in Vizgen
Updating slots in s2r1
Validating object structure for Assay ‘Vizgen’
Validating object structure for FOV ‘s2r1’
Object representation is consistent with the most current Seurat version
Object is ready!

This is the output after checking FOVs:

> vizgen.obj[[Images(vizgen.obj)[1]]]
Spatial coordinates for 3323 cells and 483 molecules
 First 10 molecules: Abcc9, Ackr1, Ackr2, Ackr3, Ackr4, Acte1
Default segmentation boundary: centroids 
 1 other segmentation boundaries present: segmentation 
Associated assay: Vizgen 
Key: Vizgen_ 
> vizgen.obj[[Images(vizgen.obj)[1]]][["segmentation"]]
A spatial segmentation for 3323 cells
> vizgen.obj[[Images(vizgen.obj)[1]]][["centroids"]] %>% str
Formal class 'Centroids' [package "SeuratObject"] with 7 slots
  ..@ cells      : chr [1:3323] "102585870299473950909222347372414346703" "102639946463719293231619392056336745817" "105321502672874017012721239064937326417" "106343835140223509995966804116631126801" ...
  ..@ nsides     : int 0
  ..@ radius     : num 80
  ..@ theta      : num 0
  ..@ coords     : num [1:3323, 1:2] 1076 1055 1081 1051 1046 ...
  .. ..- attr(*, "dimnames")=List of 2
  .. .. ..$ : NULL
  .. .. ..$ : chr [1:2] "x" "y"
  ..@ bbox       : num [1:2, 1:2] 1010 1985 7096 7442
  .. ..- attr(*, "dimnames")=List of 2
  .. .. ..$ : chr [1:2] "x" "y"
  .. .. ..$ : chr [1:2] "min" "max"
  ..@ proj4string:Formal class 'CRS' [package "sp"] with 1 slot
  .. .. ..@ projargs: chr NA

and this is the object and plot:

> vizgen.obj
An object of class Seurat 
966 features across 83546 samples within 2 assays 
Active assay: SCT (483 features, 483 variable features)
 3 layers present: counts, data, scale.data
 1 other assay present: Vizgen
 2 dimensional reductions calculated: pca, umap
 1 spatial field of view present: s2r1
> ImageFeaturePlot(vizgen.obj, features = "Slc17a7")
Warning: No FOV associated with assay 'SCT', using global default FOV

image

alikhuseynov commented 6 months ago

vizgen.obj[[Images(vizgen.obj)[1]]][["segmentation"]] A spatial segmentation for 3323 cells

ok, it does read some segmentations

It seems to be an issue here -> https://github.com/alikhuseynov/seurat/blob/vizgen_seurat5/R/preprocessing.R#L3100-L3101 or here -> https://github.com/satijalab/seurat/blob/master/R/preprocessing.R#L2778-L2780 basically sp$fov is kinda not recongnized. I don't have a bandwidth to troubleshoot right now. If you only want to try out stuff with Vizgen data, I would suggest use another public dataset with .parquet files.

alikhuseynov commented 5 months ago

@angie-john, did it finally work?