Re: Osteocyte fixation

From:tylee <>

I would be interested in a reference to staining for LDH. I'm not sure I
understand how membrane integrity could be detected in frozen sections. I am
use to seeing measurement of LDH release from cultured cells as an
indication of membrane integrity (i.e. cytotoxicity); but not in IHC
Ty Lee

-----Original Message-----
From: Hazel Stevens <>
To: john baker <>
Cc: <>
Date: Tuesday, February 13, 2001 11:00 PM
Subject: Re: Osteocyte fixation

>Hi John,
>My first thought was that you may not have many viable osteocytes after
>several days in culture due to difficulties in keeping the cells
>nourished in an organ culture type model. However this may not be the case
>if it is trabecular. The best method i can think of is to cut frozen
>sections of the biopsy and do an LDH (lactate dehydrogenase) stain. This
>picks up leaky membrane cells (dying or dead) from intact membrane
>(viable). I have never tried this technique with paraffin sections so I
>don't know if it works with those but maybe someone else has.
>Hazel Stevens,
>Tissue Engineering Science Laboratory
>6405 EBU1
>Department of Bioengineering
>University of California, San Diego,
>9500 Gilman Drive,
>La Jolla,
>CA 92093-0412,USA
>Tel: (858) 534 1765
>Fax: (858) 822 0240
>On Mon, 12 Feb 2001, john baker wrote:
>> This is for one of our graduate students here in the Orthopaedic Research
>> What is a good fixative to preserve osteocyte morphology in bone
>> The biopsy is from regenerated bone filling in a  defect in a rat femoral
>> cortex.  The biopsy is maintained in tissue culture for several days then
>> embedded in paraffin, sectioned, and stained with Alcian blue
>> The purpose of the study is to acertain the viability of the osteocytes.
>> If you know of a good way to demostrate cell viability within tissue
>> include that too.  If you require more information to help answer this
>> please let me know.
>> Thank you,  John  for Ed Hoffler
>> John A. Baker
>> The University of Michigan
>> Orthopaedic Research Laboratories
>> Histology Unit
>> 400 North Ingalls, G161
>> Ann Arbor, MI 48109-0486
>> my office 734-936-1635
>> lab office 734-763-9674

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