[Histonet] Cortical bone Image analysis fluorescent calcine measurement

[Jack Ratliff]

Try contacting Nathanael Reveal @ BIOQUANT Image Analysis Corp. They  
are also having an image analysis workshop in Nashville, TN on April  
21-23. It will cover all aspects of typical bone image analysis and  
even feature a one day session on specimen preparation using MMA  
techniques.

Jack Ratliff



On Mar 27, 2009, at 8:43 AM, Jamie E Erickson  
<jamie.erickson <@t> abbott.com> wrote:

> Hi All,
>            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
> confocal microscopy."
> 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)
> upper surface.
> 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
> bone sample.
>
> Anyway I'll get there, thanks for any help "Happy Friday".
>
> Jamie
>
> _______________________________
> Jamie Erickson
> Sr. Research Associate II M.S. HTL (ASCP)
> Department: DSMP
> Abbott Bioresearch Center
> 100 Research Drive
> Worcester, MA 01605-4341
> 508-688-3134
> FAX: 508-793-4895
> e-mail: jamie.erickson <@t> abbott.com
>
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