RE: [Histonet] Formalin vs. Saline Specimen Submission

From:"Tony Henwood"


Beware of placing specimens in saline. It is notorious for inducing
frozen section artefact in cryotomy (esp Brain and muscle biopsies).
Normal Saline, despite its name is not isotonic with cells.

Inoshita and Youngberg (Am J Clin Pathol 80(2):206-9, 1983) described
the adverse effect on skin biopsies of storage in saline solution. An
artefact, consisting of hydropic degeneration of the basal cells and
subepidermal bulla formation occurred in skin-punch biopsy specimens
immersed in normal saline. They noted similarities between the
histologic and ultrastructural findings of this artefact and of
epidermolysis bullosa simplex and pathologic hydropic degeneration, as
is seen in lupus erythematosus.

Now Susan,

My responses are below in red


Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318

-----Original Message-----
[] On Behalf Of Vicki
Sent: Saturday, 13 May 2006 2:46 AM
Subject: Re: [Histonet] Formalin vs. Saline Specimen Submission

Our frozen section room is located in the OR so we receive all of our
specimens fresh.  The Histotech assigned to the FS room is the one
responsible for placing specimens in formalin (or other appropriate
fixative for special testing) except after hours.  The OR staff then
places the specimens in formalin if FS room staff has gone for the day.
Any question on tissue for special studies the OR staff pages the tech
on call or pathologist on call for direction. We do have a grossing
station that has ventilation in case we need it . For specimens
submitted in saline....if during the day, the resident assigned to the
service for which that specimen was intended is paged immediately for
triaging of the case.  If after hours, the specimen would most likely be
placed in the refrigerator overnight and picked up first thing in the
morning.  Generally, if the specimen is to be handled right away they
generally put enough saline on the tissue to keep it wet.  If it is
going to sit longer than that, more saline would be used. Specimens that
need to be refrigerated for any reason are stored in the grossing room
refrigerator or Morgue cooler (if too large for the
fridge) and after hours,in the fridge in our lab receiving area.  After
hours, I mentioned how the OR handles their specimens for us...if there
are frozen sections involved, the pathologist on call would be paged to
come in and handle the case.  If the specimen in question comes from an
outpatient source, our lab client service center would call the tech on
call to see how to handle fresh or saline soaked specimens.  We do have
them refrigerate saline soaked specimens overnight depending on the
testing to be performed.  Phew...that was a long one !!!  Hope it
helps.. Vicki  Gauch AMCH

>>> "O'Brien, Sue"  5/11/2006 12:51:33 PM >>>
I was wondering how other hospitals handled specimens submitted from the
OR (Operating Room). Currently, our routine specimens (ones that do not
require special testing and submission) are submitted in 10% NBF
(Neutral Buffered Formalin).

Thank-you for taking the time to respond!

How are routine specimens submitted to your lab from the OR (chose one)?

In 10% neutral buffered formalin

     a. in formalin (e.g. 10% NBF)

     b. in saline

     c. fresh

     d. other (specify):

A. For specimens are received by OR in formalin:

    Is the specimen placed in formalin in the OR procedure room? Yes

If yes, how is it done?

a.       under a fume hood

b.       placed in container which has formalin - yes

c.       other (specify):

           If no, how is it done?

a.       specimen placed in container and taken fresh to another room
where formalin is added

b.       other (specify): A mix of both above (big specimens have
formalin added, small biopsies placed in formalin in theatres)

           Does the OR have a fume hood available for use (to put
specimens into formalin under it)?

B. For specimens submitted by OR in saline: NEVER

    a. How long (generally) is the specimen in saline until it is placed
into formalin?

    b. What volume of saline is used? (e.g. enough to keep moist, or
enough to submerge


     c. Who transfers the specimen into formalin? (e.g. histo or lab
tech at time of receipt, or does it

         wait until the Pathologist grosses it? If it waits for the
Pathologist, then for how long, and how

          is it refrigerated?)

    d. Are specimens refrigerated? If yes, where (e.g. in OR, in lab, in

C.  For specimens submitted fresh:

     a. how long (generally) is the specimen fresh before placement into
formalin? Depends on tests requested, some delivered directly to
Histopath on saline dampened gauge 

     b. How do you keep the small specimens from drying out? In a good
quality sealed container, biopsies should not dry out in the short time
it takes for the specimen to reach the lab.

     c. Who transfers the specimen into formalin? (e.g. histo or tech at
time of receipt, or does it

         wait until the Pathologist grosses it? If it waits for the
Pathologist, then for how long, and

         how is it refrigerated?) Depends on test requested.

     d. Are specimens refrigerated? If yes, where (e.g. in OR, in lab,
in histology).

If you receive specimens either in saline or fresh (and they are not
refrigerated immediately):

a.      How do you feel they compare to specimens that were immediately
placed into formalin? (e.g. better morphology,

      worse morphology, etc). Comparable morphology as long as saline
not used (see above)

b.      Who handles specimens submitted after hours? (e.g.
weekends/holidays; are they left at room temp, refrigerated,

      or does someone (who?) transfer them into formalin (how?). Always
placed in formalin, otherwise scientist on call attends to the specimen.

Sorry for the length, but I would really appreciate knowing how others
handle this.


Susan O'Brien, Histology Supervisor

Burdette Tomlin Memorial Hospital

Cape May Court House, NJ  08210

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