[Histonet] Cortical bone Image analysis fluorescent calcine
Jamie E Erickson
jamie.erickson <@t> abbott.com
Fri Mar 27 08:43:48 CDT 2009
Question for anyone working with image analysis on Bone..
My 1st goal: Evaluate and purchase image analysis software to measure bone
formation rate (BFR) and mineral apposition rate (MAR).
I am trying to measure the area between two fluorescent (calcine)
labels of cortical bone by image analysis.
This is standard for bone histomorphometrics in all the literature I see
but as always there is a few details that are missing.
Papers don't say what object they use, but for my mouse and rat
tibia samples, 10x won't get the whole sample in one frame? I have notes
from someone who has done this and they trace the perimeters to get the
data (with osteometric software). If I do tracing at 4x I wonder about
the accuracy. Osteometrics is the most common bone morphometric system
out there but for my cortical bone it seems a little overkill. I would
like to use Zeiss axiovision software anyone using this? I think people
are using 10-20x and trace the perimeter of the bone but I am wondering if
with certain software you can move along the perimeter of the bone without
stopping the data collection or do you have to collect multiple data sets
and sum the total to get the perimeter.
My 2nd goal: Evaluate confocal microscopy imaging of bone for reducing in
time spent grind samples.
This method was based on a short communication in Journal of
Histochemical "rapid method for the assessment of bone architecture by
Right now I am trying to determine what imaging modality I am going to use
for these measurements. My pathologists wants me to do large rough cuts
(625-1000um) from my isomet saw and see if I can measure these labels
under confocal (ie eliminating grinding) , has anyone tried this?
I can tell you that I can image these samples without a coverslip
but the samples is uneven due no grinding or placement of sample on the
slide. The theory is that I can image these samples at 10 or 20x and get
an image some microns into the sample thus eliminating the uneven (rough)
In practice I can get some areas to look great but I can't focus all the
way around the bone I lose the image focus possible due to a thicker side
of bone,, so I think this idea is not practical. Has anyone tried this?
I think going forward I will grinding samples to about 20um and mount the
samples in water based media and coverslip.
If you do this type of work please contact me I'd love to talk about
this... As always the powers to be are demanding validation studies of
this method be done yesterday and I haven't even decided on software. Not
to mention that I am new to bone work and I haven't even ground my first
Anyway I'll get there, thanks for any help "Happy Friday".
Sr. Research Associate II M.S. HTL (ASCP)
Abbott Bioresearch Center
100 Research Drive
Worcester, MA 01605-4341
e-mail: jamie.erickson <@t> abbott.com
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