Posted by
George McNamara on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Tracking-with-Metamorph-tp590031p590032.html
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalApps menu - Track Objects. As pointed out by another poster, you may
need to run MM Admin and add the dropin. See the MetaMorph help file
for information about the command. It was developed for Single
Particle Tracking (i.e. small blobs in DIC).
Another Apps menu item is Track Points, which makes it easy to do
manual tracks, analysis and graphs (always good to verify what is
going on). You can also do this with "Measure Pixels". Just go to the
first plane, click on the object to log the data (if you have the
data log open and MP dialog open), then last plane, click, Publish.
If you decide you want entire tracks, and have only one or a few
cells in a stack, and they are easily thresholded (for example, 0.1
ug/mL Hoechst or DAPI fluorescent DNA counterstained nuclei), you can
use Integrated Morphometry Analysis / Measure Objects with "loop for
all planes". I prefer Measure Objects (legacy dropin measobj), set up
threshold, classifier(s), measurements (at least X, Y, plane!), and
set up the inner loop journal to add the measured result image into
the bottom of a stack. If you have "show centroid" on (Preferences),
you will get a set of images that have the object(s) with their
centroids highlighted.
Hint: I recommend NOT doing "calibrate distances" (Measure menu).
Easier to log the raw image data - pixels - to Excel, understand the
data, then convert from pixels to um at the end. If you are not
rigorous about applying calibrate distances, some images may be in um
(or lightyears, etc) and others may be pixels.
If your cells have high contrast (i.e. fluorescent nuclei) you can
use Process menu: Stack Arithmetic: Max, so see all pixels covered
("temporal area maps" see
http://home.earthlink.net/~geomcnamara/temporal_area_maps.htm I
typically make a binary stack first, then do TAM). You can likewise
use Stack menu: Topographic Surface Maps, to see first time point
covered. If you make the TAM map, you may be able to put a line ROI
on the beginning and end of the track, and use the Measure Regions
table to get the distance you want.
Your friendly neighborhood MetaMorph sales rep can show you how to do
all these items. you can also call UIC/MDS/MDS for help.
best wishes,
George
At 07:41 AM 8/16/2007, you wrote:
>Search the CONFOCAL archive at
>
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal>
>Dear All,
>
>We are trying to analyze cell motility using Metamorph
>software. We could not find the way to measure the net
>displacement of the cell, i.e., the straight line
>distance between the first point to the last point of
>the cell track traced. I will be thankful if anybody
>could help.
>
>Regards,
>
>Nishigandha Naik
>
>
>
>
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http://mobile.yahoo.com/go?refer=1GNXICGeorge McNamara, Ph.D.
University of Miami, Miller School of Medicine
Image Core
Miami, FL 33010
[hidden email]
[hidden email]
305-243-8436 office