Hi,

I’d like some input on whether I’m using various geometric morphometric 
programs correctly or not. I’m a research technician working on a geometric 
morphometric project on seed beetles, and I’ve mainly taught myself how to 
use all these programs by reading various handbooks and such. Since there 
are a lot of steps, I want to make sure I’m not missing anything or doing 
something incorrectly. Here’s how I’ve been doing it, starting from placing 
landmarks all the way to running statistical analyses:

1. I use tpsUtil32 to create a tps file of all the photos I want to 
landmark using the “Build tps file from images” operation. I hit “Input” 
and choose an image file from the folder where all my images are, then I 
hit “Output” and create a name for the tps file. I then select “Setup” and 
include all the images I need, then hit “ok”
2. Then I upload this file to tpsDig264 and place the landmarks first, then 
the semilandmarks, for each specimen. I use the “Digitize landmarks” tool 
for landmarks, and the “Draw background curves” tool for semilandmarks. I 
make sure every specimen has the same number of landmarks and 
semilandmarks, using the “Resample curve” option to keep the number of 
semilandmarks consistent for each curve
3. Once I’m done landmarking, I open the tps file using Notepad and change 
the number after “ID=” to the unique specimen IDs I’ve given to each 
specimen
4. Then I use CoordGen8 and upload the tps file using the “load tps, 
convert curves, create SLM proto” option. I save the protocol file, then do 
a “SemiLandMark Alignment” and upload the protocol file to align all the 
landmarks and semilandmarks. Then I hit “save all”
5. The resulting file then has to be formatted for MorphoJ, which involves 
adding the specimen IDs to the beginning of each block of coordinates that 
corresponds to that specimen, deleting the centroid size at the end of each 
block of coordinates, making sure a paragraph mark separates each 
specimen’s block of coordinates, and adding a comma followed by a space 
between every number
6. Once the formatting is done, I upload this file to MorphoJ 1.07a by 
selecting “create new dataset” and selecting the data as 2 dimensions, 
selecting “yes” for object symmetry if applicable, and choosing the file 
type as “text.” Then I click “new procrustes fit” and line up the landmarks 
if there is object symmetry. I look for any outliers, generate a covariance 
matrix, extract classifiers from my specimen ID, and run whatever analyses 
I need to (PCA, CVA, ANOVA, etc.)

Are there any steps I’m missing or things I’m doing incorrectly?

Thank you in advance,
Lydia Irons

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