[Histonet] Histozymmography

From:"Michelle McDonald"

I would like to hear from anyone who has any experience with histozymmography on bone tissue sections.

We are in the process of optimising a technique to analyse MMP activity in adult rat growth plates in bone samples.

We have been able to cut frozen sections of decalcified samples but as yet have not perfected frozen sections of undecalcified samples even using a tungsten carbide blade. 

We are also currently coating slides with fluorescently labelled gelatin or casein dissolved in a 1% agarose in 50mM Tris Ph 7.2 solution at 37 degrees.
It is difficult to get a consistent thick coating on the slide and under the microscope these coatings look grainy. 

If anyone has any suggestions on how we can optimise these techniques we would be greatly appreciative

Thank you

Kind Regards


Dr Michelle McDonald B.MedSci, PhD
Research Officer
Orthopaedic Research Dept.
The Children's Hospital Westmead.
Westmead NSW 2145
Ph. +612 9845 1451
Fax. +612 9845 3078

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