Hi
I suspect that all your advisor wants to do is to get the relative
amounts of each of the leukocytes.
To do this select an area of the blood smear where the red blood cells
are separated a little. Avoid clumps where the smear will be thick or
the tail ends of the smear where the white cells may be clumped and
distorted.

Ideally the red blood cell should be salmon pink. If they are red then
all the colors that you have in a description of the white cells will be
shifted towards the red, if the red blood cells are slate blue then
similarly all the colors will be shifted towards the blue.

Start by counting the white cells in the first field. 
Move the slide one field over say to the right. Count these cell types. 
Then move one field down and count. 
Then one field to the right and count.
Then up one field and count.
Repeat this procedure until you have counted at least 200 preferably
more leukocytes.
If you get to the end of the slide or in an undesirable area during
moving then move down one field and repeat this but moving to the left
etc.
This is the battlement method of counting and will give you a
differential count of leukocytes.
The more leukocytes you count the more accurate your percentages. 
You may have trouble in finding basophils as they are only present in
the order of 0.5 to 1%. 
Hope that this helps
Barry

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Poteete,
Jacquie A.
Sent: Tuesday, September 26, 2006 2:39 PM
To: Ford Royer; Missy; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] WBC counts

Good luck finding a hemacytometer and/or any calibrated pipettes.  Ours
ended up in the display of "antique instruments and equipment".  That's
the best place for them (speaking as an old dinosaur who actually
learned how to use them).

Jacquie Poteete MT(ASCP)QIHC
Lead Medical Technologist, IHC Laboratory
Saint Francis Hospital, Tulsa, OK

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ford
Royer
Sent: Tuesday, September 26, 2006 2:24 PM
To: 'Missy'; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] WBC counts


Melissa,

It sounds like your advisor wants a "differential" WBC if you are
attempting to do it on a stained peripheral blood smear.  To do this you
would need some type of multi-key manual differential counter (example:
http://stores.implex.net/minnesotamedical/product_info.php?cPath=205_174
_176
&products_id=1726)

To do this you assign a key on the counter to a specific white cell
(i.e.
nuetrophils) and do the same for the rest of the keys with the different
types of white cells.  You then scan the slide and count the different
white cells (punching the proper key for each) until you reach a total
count of 100 cells (pay no attention to the red cells).  Your
differential would then be the number of specific cells counted express
as a percentage.  Example, you counted 62 nuetrophils and 38 lymphocytes
= 62% neturo & 38% Lymphs respectively.

To do a total WBC, you would have to have a hemocytometer (what Tim was
talking about), calibrated hemo pipettes, a lysing reagent, normal
saline diluent, and WHOLE blood.  You would dilute the whole blood to
the proper dilution with normal saline, add the lysing reagent to
destroy the red cells, load the hemocytometer, and count all the white
cells you see.

Or, as Tom suggested... give the assignment to the Hematology
Department.
;-)

Ford M. Royer, MT(ASCP)
Histology Product Manager
Minnesota Medical, Inc.
7177 Madison Ave. W.
Golden Valley, MN 55427-3601
CELL:  612-839-1046
Phone:  763-542-8725
Fax:  763-546-4830
eMail:  froyer@bitstream.net

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken,
Tim
Sent: Tuesday, September 26, 2006 11:21 AM
To: Missy; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] WBC counts

Is this some kind of project for a class? If not, hand it over to
hematology. If so, get a medical technology text book and look up blood
cell counting. To do it right requires special counting slides that are
etched with grids. 


Tim Morken

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Missy
Sent: Tuesday, September 26, 2006 7:25 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] WBC counts

Hi all...
We have never done a manual WBC count before and my advisor has asked me
to get a count of WBCs (primarily nuetrophils and lymphocytes) in
wright's stained peripheral blood smears.  What is the general protocol
for a manual count? Do you count WBC's inb relation to RBCs or just
WBCs, the whole slide, or just representative sections?

Melissa

P.S - this is for rat sample, not human
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