[Cytometry] about absolute WBC count from ISHAGE strategy and from the hematology analyzer

KANCHAN JESWANI kanchan.jeswani at hotmail.co.uk
Fri Mar 2 01:09:28 EST 2012


Hi Rakesh!
Thanks !
I am doing  cord blood CD34 ,stem kit on FC500 since5 yrs we do preprocessing and post processing CD34 enumeration.Most of our samples show a higher WBC count on the hem analyzer which I presume is due to counting of viable and nonviable cells and also NRBC.I have nt derived the percentage difference but will do it ASAP to see our findings.
Our post processing CD34 yield is normally lower or same as preprocessing CD34.but sometimes it is more than preprocessing since the time  the cord blood started processing with SEPAX?
Wht is your experience?
Thks
Kanchan jeswani
Clinical scientist
Dubai cord blood and research centre,DHA
Dubai
The  
 

Subject: Re: [Cytometry] about absolute WBC count from ISHAGE strategy and from the hematology analyzer
From: rnayar at uhnres.utoronto.ca
Date: Thu, 1 Mar 2012 10:07:13 -0500
CC: jmendoza.rmt at gmail.com; rob.sutherland at utoronto.ca; cytometry at lists.purdue.edu
To: kanchan.jeswani at hotmail.co.uk



Hi Kancham,
You're right, the majority of cases you'll find a higher WBC from a hematology analyzer.  We've been analyzing chords for greater than 15 years and found a very good correlation between our hem analyzer and CD45 derived WBC on our flow instruments.  I went through the last two months of data (600+ samples) and found that there was on average a 6% difference in counts between the two instruments, with 87% of those counts were higher on the hematology analyzer.  Seven samples fell out of exceptable range.  When we fall out of range, at greater than 15% difference, it tends to be samples that have either poor viabilities or a lot of platelet adhesion.  It should be noted that we use our hematology analyzer as an external control for single platform counts (CD34, CD4 etc).  Our ISHAGE kit recommends we run duplicates and it's very easy to make the same mistake twice or six times when your batch analyzing.
Cheers,Rakesh
Rakesh (Casey) NayyarCytoquest Corporation (ISO 15189 Certified)Flow Cytometry ServicesToronto General HospitalThe Norman Urquart Wing, Room 11NU-1115200 Elizabeth Street, Toronto, Ontario, M5G 2C4Telephone:416-946-4501 ext. 5237www.cytoquestcorp.comrnayar at uhnres.utoronto.ca
On Feb 29, 2012, at 12:29 PM, KANCHAN JESWANI wrote:Hi
Is it not true that Leucocyte count by single platform ISHAGE protocol is lower than WBC count from the hematology analyser?
Our experience from cord samples using stem kit
Rgds
Kancham 

Date: Wed, 29 Feb 2012 09:09:42 +0300
From: jmendoza.rmt at gmail.com
To: rob.sutherland at utoronto.ca
CC: Cytometry at lists.purdue.edu
Subject: Re: [Cytometry] about absolute WBC count from ISHAGE strategy and from the hematology analyzer

Hi Rob,

Thanks for the input .

In my previous job, less than 5000 WBC/uL count difference between WBC
counted on the Hematology analyser and CD45 on the flow  is acceptable
because they have their validation paper to support it.

Our analyser does NRBC count and corrects the WBC if it needs to. I totally
agree with you that CD45+ is more accurate and we are using single ISHAGE
protocol.

Regards,
Jen

On Wed, Feb 29, 2012 at 5:28 AM, D. Robert Sutherland <
rob.sutherland at utoronto.ca> wrote:

Hi Adamari,
re bone marrow samples, I would expect the CD45+ cell count from a single
platform ISHAGE protocol, provided the assay is performed properly, to be
more accurate than the WBC count from a hematology analyser due in the main
to the variable presence of nRBCs in the sample. NRBCs are not the only
source of discrepancy here, but for BM samples, the most likely one.
One question; why are you counting CD34+ cells in BM samples? BM samples
are still occasionally used for transplant, but the CD34+ cell count is
irrelevant here. Traditionally for transplants using BM samples, a WBC
count is performed and the bag of marrow is weighed. There are no published
studies to my knowledge that correlate CD34+ cels with engraftment in the
BM setting, and if there were they would be meaningless given (among a host
of other reasons) the highly heterogeneous characteristics of BM CD34+
cells compared with say CD34+ cells in mobilized PBSC samples.

Re Jen's response, nRBCs can account for up to 20%  of the nucleated cell
count in cord blood samples apparently. Again, the CD45+ cell count will
give you a more accurate WBC count than a hem analyser on some CB samples.
What does "5.0x10e3" refer to?
best
rob

D. Robert Sutherland
University Health Network/Toronto General Hospital

On 2012-02-28, at 3:54 PM, Jen Mendoza wrote:

Hello everyone,

I was just about to ask the same question for the same reason but for
cord blood as sample.

I know below 5.0x10e3 is acceptable but what's the basis, I don't know,
 I can't find anything on the net to support it. I'm sure I'm not looking
on the right path, so if anyone has info. please share.

Thanks ahead,
Jen

Sent from my iPhone, please excuse brevity and spelling mistakes.


On Feb 24, 2012, at 8:34 PM, Adamari Lopez <ALopez3 at cbcsf.org> wrote:

Good morning Flowers:

I've seen differences between WBC from the hematology analyzer and
FACSCalibur , depending of the type of samples. The bone marrow is the one
that give me the biggest difference because of the nRBCs, how much is it
accepted as difference for this type of samples ?
We don't get that many bone marrows here  to make a research, but I
would like to set up an standard of acceptability for this parameter.
We use the hematology analyzer just for checking if  a dilution it's
needed before staining the samples for CD34 enumeration.
Any comment will be helpful.
Thanks,

Adamari Lopez

QC-Flow  Lab
Community Blood Center of South Florida


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