Studying cell metabolism from frozen tissue samples - Is this even possible? - (Dec/20/2006 )
My lab frozen human cornea explants with tissue tech, alcohol and dry ice. This procedure were done two years ago. Now, we wanna thaw the samples to study their metabolism, putting riboflavin and UV light and analyse collagen behavior. Is that possible?
-Suy-
QUOTE (Suy @ Dec 21 2006, 02:32 AM)
My lab frozen human cornea explants with tissue tech, alcohol and dry ice. This procedure were done two years ago. Now, we wanna thaw the samples to study their metabolism, putting riboflavin and UV light and analyse collagen behavior. Is that possible?
you can analyze some aspects of status of collagen when it was frozen, let´s say posttranslational modifications, structure of fibres, interaction with protein complexes a.s.o. by staining methods; "metabolism" in situ in the way to measure dynamic physiological reactions won´t work;
what riboflavin and UV for?
-The Bearer-
QUOTE (kosmodrom @ Dec 21 2006, 05:27 AM)
QUOTE (Suy @ Dec 21 2006, 02:32 AM)
My lab frozen human cornea explants with tissue tech, alcohol and dry ice. This procedure were done two years ago. Now, we wanna thaw the samples to study their metabolism, putting riboflavin and UV light and analyse collagen behavior. Is that possible?
you can analyze some aspects of status of collagen when it was frozen, let´s say posttranslational modifications, structure of fibres, interaction with protein complexes a.s.o. by staining methods; "metabolism" in situ in the way to measure dynamic physiological reactions won´t work;
what riboflavin and UV for?
Thirst of all, thank you so much for the answer. I really appreciat that.
The corneas have a pathology called keratoconus, that is a cornea ectasia (the cornea starts to get thinner, and usually the patient has to do a corneal tranplantation to get their vision back. With riboflavin and UV light, we try to do cross link the corneal collagen fibers, to see if the cornea get stronger and the pathology doens't progress anymore.
-Suy-