Bone and Cartilage Cryosectioning and Staining

From:Lora Huang

Dear HistoNet,
    I usually do Affymetrix GeneChip and spotted array work, but I have an
investigator with a question that I could not answer, so I thought I would
try everyone here.  This investigator would like to induce a defect in the
tibia of rabbits, wait a specific time to allow for healing, and then
section the bone for use on a Laser Capture Microscope.  The goal would be
to isolate the RNA of the different cell populations present during the time
line and then use that RNA on Afftmetrix GeneChips to check for differences
in expression.
    I tried doing some background research and found a protocol from DeCal
using their ImmunoCal solution to decalcify the bone and they say that this
will preserve RNA.  Does anyone know if this will work?
    My other question is how to best stain the bone and cartilage once it
has been sectioned?  I just need enough stain to be able to differentiate
between the different cells without killing the RNA.  Any help anyone might
be able to lend would be GREAYLY appreciated!!  I have only ever sectioned
organs and tumor tissue, so I'm pretty much helpless.

Thank you!!!
Lora Huang
Univ of Iowa DNA Facility
----- Original Message -----
From: "HistoNet Server" 
To: "Lora Huang" 
Sent: Thursday, January 16, 2003 9:59 AM
Subject: re: subscribe

> Your address has been added to the addresses that comprise this  Listserv
> List.
> Welcome to HISTONET. This is an electronic mailing list for the exchange
> information pertaining to histotechnology and related fields.
> It contains useful information about how to use the list and what to do if
> experience problems. It also includes some basic rules for email etiquette
> (Netiquette) which will be helpful to those who are new to this form of
> communication.
> A list server is a computer that runs software which will receive incoming
> electronic mail (email) messages and reroute them automatically to
everyone on
> the subscriber list. Email uses the vast expanse of the Internet to allow
> almost instantaneous communication between networked computers around the
> world. Our system uses the LISTSTAR software from Quarterdeck Corporation
> (California) and can currently send about 30 messages a minute.  With the
> present number of subscribers, we are processing about 10,000 outbound
> messages a day.
> Anyone interested in research or clinical applications of histology,
> immunohistochemistry, in-situ hybridization pathology, and electron
> may find Histonet informative and useful. Currently, there are more than
> subscribers from all over the world. Subscribers include hospital
> from major urban centers and obscure remote locales, university
> botanists and the employees of commercial laboratories, government
> veterinary facilities and a wide variety of commercial industrial
> The list is run by Linda R. Margraf, M.D. and Herb K. Hagler, Ph.D. using
> hardware and software owned by the University of Texas Southwestern
> School, Department of Pathology in Dallas, Texas. If you have any
questions or
> problems with Histonet please contact Linda Margraf at
> This server, unlike many systems, uses ONLY ONE ADDRESS to send commands
> the computer and to post messages. The server will recognize commands sent
> the SUBJECT line of the message and only when they are spelled exactly as
> listed below. Anything not identified as a command will be circulated to
> EVERYONE on the list.
> The following is a list of commands the server recognizes:
> subscribe
>  Your address will be added to the list of subscribers. You will then be
> to send messages to this list that will be forwarded to all other list
> subscribers. You will begin to receive all messages sent to the list by
> subscribers.
> subscribe digest
>  Your address will be added to the list of subscribers who receive a
> instead of each forwarded message. A digest is a compilation of all the
> messages received in a 24 hour period. It is sent to the digest
> every night after midnight. Digest subscribers can post and respond to
> messages the same as "real-time" subscribers.
> digests
>  A list of available digests will be returned to you. Histonet stores old
> messages as daily digests for approximately three  months. To read
> messages, copy the list of available  digests, mark the dates of interest
> return it to the server.
> unsubscribe
>  Your address will be removed from the list of subscribers.
>  You will no longer be able to send messages to the members
>  of the list.
> help
>  A list of the commands recognized by the server will be   returned to
> You may post any questions you wish pertaining to histology, pathology,
> in-situ hybridization, immunohistochemistry etc. Equipment and reagent
> evaluations, laboratory management issues, government regulations, and job
> opportunities are all appropriate topics. The University asks that we
> the use of its hardware and software to business purposes only (occasional
> jokes do slip through but PLEASE use restraint). Vendors and those with
> commercial interests in histology products are welcome contributors
> we ask that blatant advertisements be avoided at all times. It is fine to
> refer to product that your company produces if it is pertinent to a topic
> being discussed on the list. Unsolicited advertisements are poorly
> by the members and you will likely receive a number of negative comments
> you overstep the boundaries. Please contact Linda Margraf at
> if you are not sure about the
> if a message you wish to post.
> It is most helpful to the list members if you post your responses to
> to everyone on the list and not just as a personal reply to the person
> the question. That way duplicate messages are minimized and we all learn
> each other's comments.
> Likewise, if you post a question and get a number of responses back
> to you, it is helpful to everyone if you could send out a summary of the
> replies you got to Histonet.
> Please avoid abbreviations unless they are explained in your message. For
> example: immunohistochemistry (IHC). This list circulates to a wide
variety of
> individuals and what seems obvious to you may have no meaning on the other
> side of the world.
> Please sign your letter and include your institution or affiliation and
> location. Not all email systems have headers which identify the sender.
> Do use the subject line to indicate the topic of your message.
> DON'T USE ONLY CAPITAL LETTERS -it is considered shouting.
> Please send questions and problems about the list directly to Linda
Margraf at
> and don't circulate them to the >850
> subscribers on the list. Be careful when sending commands to the server to
> the command in the SUBJECT LINE and spell it correctly.
> Please do not send images as attachments with your message. We can now
> images at our web site ( To have an image
> send it to Herb Hagler at
> es

<< Previous Message | Next Message >>