instructions for old Coulter Counter model CBC5?

Hello all, could use someone's help here. We've managed to pick up a
used countertop Coulter Counter, model CBC5, which, while old
("obsolete" per Beckman-Coulter) looks like it would suit our lab's
needs just fine. We need to do an approximate WBC on blood and marrow
samples before setting them up for culture at a particular WBC/volume
ratio.

Unfortunately, the machine did not come with a manual, so we have no
idea how to calibrate it and run samples appropriately. Beckman claims
they have no manual available, tho I'm sure they do have one in a file
somewhere, they just want us to buy a newer model!

Does anyone out there have a CBC5 *with* a manual, who might be
willing to copy or fax it to us?

It would be much appreciated!

Anita

Well this B/C instrument is so old I have never heard of it. There are a
lot of newer small used and referbished instruments out there that would be
cheap and a lot less trouble. But if you are the type that likes to flay
your back with chains or just don't have the money (in which case what you
doing in this biz?). ..... That said, most MTs with some experience could
probably create their own manual. More troublesome is what do you use for
calibrator and reagent? Calibrators, controls and reagent are not without
expense. The manufacture generally matches the calibrator targets agianst
the instrument. Here you have an unsupported instrument with no reference
values. You can't (or shouldn't be) running bone marrow solutions on a
CBC5. You need an instrument with a flourencent flow cytometer capability.
E.g. how you know your not counting NRBC? Just my opinion, don't claim to
know everything.
<ahawkins@cancergenetics . com > wrote in message
news:1186585582.601647.289720@g4g2000hsf.googlegroups . com ...
> Hello all, could use someone's help here. We've managed to pick up a
> used countertop Coulter Counter, model CBC5, which, while old
> ("obsolete" per Beckman-Coulter) looks like it would suit our lab's
> needs just fine. We need to do an approximate WBC on blood and marrow
> samples before setting them up for culture at a particular WBC/volume
> ratio.
>
> Unfortunately, the machine did not come with a manual, so we have no
> idea how to calibrate it and run samples appropriately. Beckman claims
> they have no manual available, tho I'm sure they do have one in a file
> somewhere, they just want us to buy a newer model!
>
> Does anyone out there have a CBC5 *with* a manual, who might be
> willing to copy or fax it to us?
>
> It would be much appreciated!
>
> Anita
>

Kuhnfucius makes a really good point regarding calibrators. How will you
know if you're in range if you can't buy commercial calibrators any
more?

* findarticles . com /p/articles/mi_m3230/is_n13_v21/ai_8263509/pg_57

* w w w .rndheme . com /index.php?page_id=id_showproduct&product_id=CBC7
- although this company seems to offer calibrators for this
instrument.....

* w w w .msb-inc . com /displayParts.cfm?pageId=41&productId=206 - this
place even services them and has parts....

This article on the CBC5 is from 1989. That makes this machine nearly 20
years old. That is the dark ages when it comes to instrumentation. Also,
the reagent question is a good point. All Coulter machines need diluent.
I even know that and haven't done Hematology since 1988 (our lab didn't
use a CBC5).

All in all, it seems as if you may be dead in the water. The NRBC
question is a good point also, as I seem to remember something from
student days about correcting for NRBC's if you see them. Isn't bone
marrow kind of chunky? Wouldn't it clog up your instrument? Since you
are talking about culturing, why do you need a count before culture?
I've never heard of that before. We rarely get bone marrow cultures.

Are you doing this on human specimens? For research? If these are animal
specimens then your calibrator problem gets even MORE complicated.

Judy Dilworth, M.T. (ASCP)
Microbiology

"kuhnfucius" <krk4567@comcast . net > wrote in message
news:Z_qdnR9gCY8H-CPbnZ2dnUVZ_sejnZ2d@comcast . com ...
> Well this B/C instrument is so old I have never heard of it.

"JEDilworth" <bactitech@nospamhortonsbay . com > wrote in message
news:8K6dnYPghtsvHiPbnZ2dnUVZ_hCdnZ2d@buckeye-
>
> All in all, it seems as if you may be dead in the water. The NRBC question
> is a good point also, as I seem to remember something from student days
> about correcting for NRBC's if you see them. Isn't bone marrow kind of
> chunky? Wouldn't it clog up your instrument? Since you are talking about
> culturing, why do you need a count before culture?

Since you have no manual and no way to calibrate, the number that you get
out of the analyser could be referring to any one or more of:

- The white cell count
- The red cell count
- The platelet count
- The nucleated cell count
- The total number of small things in the sample
- Room temperature
- Any randomly generated number.

Personally I wouldn't touch this set-up with a barge pole!

Yup, that's true!

Judy D.

"Manky Badger" <you.must@be.joking> wrote in message
news:FN-dnWrTwvhyU1zbRVnyjgA@giganews . com ...

> Since you have no manual and no way to calibrate, the number that you
> get out of the analyser could be referring to any one or more of:
>
> - The white cell count
> - The red cell count
> - The platelet count
> - The nucleated cell count
> - The total number of small things in the sample
> - Room temperature
> - Any randomly generated number.
>
> Personally I wouldn't touch this set-up with a barge pole!
>

Kuhn, Judy - thanks for your replies, much appreciated. I can see tho,
that I didn't explain myself very well, since you're both telling me
basically that I don't want to be doing what I DO want to be doing :)
So let me try to go back and say more clearly why we want to do this,
and we'll see if maybe I really am trying to beat a dead horse into
getting up and walking...

We are a cancer cytogenetics lab specifically, not a general path or
hem lab, and unfortunately don't have one close by to ask for advice.
Basically what we do is receive blood and bone marrow specimens from
physician's offices in order to perform G-band karyotype analysis and
FISH for cancer-specific (mostly leukemia and MDS) abnormalities. We
culture the specimens short-term in special medium to encourage the
cells to divide so that we can examine the chromosomes at the
microscope after harvest. This is tissue-culture, not microbiological
culture (at least we try really hard not to get any microbial
growth!). The cells grow best in culture if they're set up at a
moderate density, yet our patient specimens as recieved vary several
orders of magnitude in their nucleated cell density, from new-
diagnosis CLLs or acute leuks to post-treatment or rule-out-MDS
pancytopenias.

Surely all these bloods and marrows are having cell counts done on
them as well, but not here, and the results are not available to us
unless we chase them down, one dr's office at a time. Plus, we often
need to set up cultures right away, and can't delay wating for someone
else's results to be communicated to us.

So right now we dilute out a bit of sample in acetic acid to lyse,
then count manually on a hemacytometer. This is fine for a few
specimens a day, but our volume is going up quickly, and we're
spending ridiculous amounts of time daily counting these things.
Especially since the countable range is so narrow that most specimens
we need to either re-dilute, or concentrate, in order to be able to
count a decent number.

We want something like a CBC5 to do simple semi-automated WBCs, with
some reasonable dilution range, and +/-20% accuracy (remember we're
just trying to adjust our culture inoculation volume to within a rough
range, we're not reporting these numbers at all!). There may indeed be
other small more-recent machines that we could turn to, but for now, I
just want to find out if this one that we have is going to be usable
for us (or not).

So now on to your specific comments:
Diluents and calibrators are available, not from B-C but from second-
market sources; Judy, thanks for an additional source that I hadn't
found yet.

No doubt an experienced MT could figure out some reasonable
approximation of a manual, but again, no MTs here, all cytogenetic
tech specialists and not generalists, alas.

I'm not going to worry about a few percent of nucleated reds, since no
doubt we're counting them already with the hemacytometer :) and flow
capability would be a lot more information than we need! Most of these
spec are split for flow before they send part on to us anyway; in an
ideal world, they would send us their cell counts before we needed to
set up our cultures, but in practice it ain't gonna happen.

We do dilute the marrows and let the chunks settle out before putting
them in for counts; we almost never see spicules in the hemacytometer
chambers. I hope clogging won't be a problem with the counter, but
that's one of the things I'd hoped a manual would settle.

And yes, these are human specimens. I did grow up a blood from my
horse for chromosomes once, and have done a few mouse and rat samples
thru the years... but these are just ordinary 2-legged animals :)

So... to get back to the point... the orginal actual question...

Does anyone out there have a CBC5 *with* a manual, who might be
willing to copy or fax it to us?

Thanks
Anita

<ahawkins@cancergenetics . com > wrote in message
news:1187121607.040147.134770@g4g2000hsf.googlegroups . com ...

> Surely all these bloods and marrows are having cell counts done on
> them as well, but not here, and the results are not available to us

>Most of these
> spec are split for flow before they send part on to us anyway; in an
> ideal world, they would send us their cell counts before we needed to
> set up our cultures, but in practice it ain't gonna happen.

Why on earth not?
In our modern era of making results available electronically, you should
have the facility for results to be available pretty much instantly.


> So... to get back to the point... the orginal actual question...
>
> Does anyone out there have a CBC5 *with* a manual, who might be
> willing to copy or fax it to us?

Doesn't look like it.
You're probably asking the wrong people - try the charities that supply
unwanted medical equipment to third world countries. Without wishing to
sound rude or patronising, they are probably the only people who would deal
with a blood counter that went out of date over fifteen years ago.

Anita said: > We are a cancer cytogenetics lab specifically, not a
general path or
> hem lab, and unfortunately don't have one close by to ask for advice.
> Basically what we do is receive blood and bone marrow specimens from
> physician's offices in order to perform G-band karyotype analysis and
> FISH for cancer-specific (mostly leukemia and MDS) abnormalities.

Even assuming all of your givens, your calibration WILL change from day
to day. You absolutely have to have somewhere to start from. You can't
just pick numbers out of the air. You have to know the machine is
functioning.

Don't you have some sort of accreditation standards that must be met for
your type of lab? CAP is a very marketable accreditation, but I'm not
sure if they are the ones used for labs such as yours. I used to market
laboratory services. Do the physicians who use you realize that you're
charging them for specimens to be standardized on an ancient analyzer? I
know you're not technically reporting from this machine, but you're
still using it to standardize your inoculum. You owe your patients and
clients a reasonable effort to stay current with equipment.

Spend some money and do this with it:

1. Buy an analyzer that has been made within the past five years that
comes with a manual. I'm sure there are even used ones out there in this
age range, as many labs merely lease them. Contact the big companies for
information. Get reps in and find out what your competitors are using.

2. Hire a medical technologist who knows what they're doing in
hematology. If you can't do that, advertise for someone who has
hematology experience to do side work to at least set up your newish
analyzer and show you all how to calibrate it and use it AND do the QC
for it. Have this person show you how to tell when you need to call in
someone to fix it.

3. Set up a daily QC chart showing all the parameters and ranges of your
calibrators to show whether you are in range. Figure out your 2 SD +/-
ranges and do things like they should be done.

4. I hope you're using sterile pipettes to go into your bone marrow
samples. Otherwise, you are introducing bacteria into your tissue
cultures that could possibly mess up your FISH results.

5. Set up communication with your physician's offices to get results
that you need with the labs they use.

Could you set up something to use a spectrophotometer to get your
density, rather than a cell counter? Our lab does FISH and karyotyping,
but I don't do any of that stuff and am clueless. I do know that we
don't have any twenty year old equipment in our laboratory.

No offense, Anita, but the way you describe your operation it sounds
like a garage lab. If you're competing with the big boys (believe me,
every lab is competing with every other one - I've been there) you need
to use what your competitors are using. I really doubt that they are
using 20 year old equipment. Where are you going to find parts when your
old thing breaks down? It's just not worth the hassle.

Judy Dilworth, M.T. (ASCP)
Microbiology

On Aug 14, 9:43 pm, "JEDilworth" <bactit...@nospamhortonsbay . com >
wrote:

<snip>
> 5. Set up communication with your physician's offices to get results
> that you need with the labs they use.
>
> Sounds like this is what you need to do, it seems like you are bending over backwards to try and get cell counts that are already done by your refering centres. make a cell count a REQUIRED part of submitting a sample for analysis. If you don't get compliance from time to time, phone them and results should be almost immediately availavble. the time and trouble making a few phone calls seems like a no brainer rather than try to resurect an obsolete piece of equipment.
Matbe we're missing something, is there some special reason you can't
get the results from the specimen source?

"rickh" <harrison6723@rogers . com > wrote in message
news:1187196482.573417.23290@q3g2000prf.googlegroups . com ...
> On Aug 14, 9:43 pm, "JEDilworth" <bactit...@nospamhortonsbay . com >

Mind you, it's good to see a thread here more than two posts long that isn't
about iron :o)

Not a dead horse we can learn a lttle about karyotyping. If some of the
questions are blunt, it is just that I am probing to collect information. I
always wanted to get into cell cultures and bio-reactors (oh well...there is
still power ball).
I know what is charged ($) for FISH and karyotyping. It is expensive.
Which brings up question why your facility is cheaping out on a good cell
counter (note I do not state expensive). MDS is probably the most common
diagnosis on chromosome analysis request. Most are going to be greatly
diluted with blood (non-diagnostic). How do you know you have enough blast
cells (e.g. CD-34) for culture (hit or miss?). Do any proficiency testing
for your hemacytometer use?

ahawkins@cancergenetics . com > wrote in message
news:1187121607.040147.134770@g4g2000hsf.googlegroups . com ...
> Kuhn, Judy - thanks for your replies, much appreciated. I can see tho,
> that I didn't explain myself very well, since you're both telling me
> basically that I don't want to be doing what I DO want to be doing :)
> So let me try to go back and say more clearly why we want to do this,
> and we'll see if maybe I really am trying to beat a dead horse into
> getting up and walking...
>
> We are a cancer cytogenetics lab specifically, not a general path or
> hem lab, and unfortunately don't have one close by to ask for advice.
> Basically what we do is receive blood and bone marrow specimens from
> physician's offices in order to perform G-band karyotype analysis and
> FISH for cancer-specific (mostly leukemia and MDS) abnormalities. We
> culture the specimens short-term in special medium to encourage the
> cells to divide so that we can examine the chromosomes at the
> microscope after harvest. This is tissue-culture, not microbiological
> culture (at least we try really hard not to get any microbial
> growth!). The cells grow best in culture if they're set up at a
> moderate density, yet our patient specimens as recieved vary several
> orders of magnitude in their nucleated cell density, from new-
> diagnosis CLLs or acute leuks to post-treatment or rule-out-MDS
> pancytopenias.
>
> Surely all these bloods and marrows are having cell counts done on
> them as well, but not here, and the results are not available to us
> unless we chase them down, one dr's office at a time. Plus, we often
> need to set up cultures right away, and can't delay wating for someone
> else's results to be communicated to us.
>
> So right now we dilute out a bit of sample in acetic acid to lyse,
> then count manually on a hemacytometer. This is fine for a few
> specimens a day, but our volume is going up quickly, and we're
> spending ridiculous amounts of time daily counting these things.
> Especially since the countable range is so narrow that most specimens
> we need to either re-dilute, or concentrate, in order to be able to
> count a decent number.
>
> We want something like a CBC5 to do simple semi-automated WBCs, with
> some reasonable dilution range, and +/-20% accuracy (remember we're
> just trying to adjust our culture inoculation volume to within a rough
> range, we're not reporting these numbers at all!). There may indeed be
> other small more-recent machines that we could turn to, but for now, I
> just want to find out if this one that we have is going to be usable
> for us (or not).
>
> So now on to your specific comments:
> Diluents and calibrators are available, not from B-C but from second-
> market sources; Judy, thanks for an additional source that I hadn't
> found yet.
>
> No doubt an experienced MT could figure out some reasonable
> approximation of a manual, but again, no MTs here, all cytogenetic
> tech specialists and not generalists, alas.
>
> I'm not going to worry about a few percent of nucleated reds, since no
> doubt we're counting them already with the hemacytometer :) and flow
> capability would be a lot more information than we need! Most of these
> spec are split for flow before they send part on to us anyway; in an
> ideal world, they would send us their cell counts before we needed to
> set up our cultures, but in practice it ain't gonna happen.
>
> We do dilute the marrows and let the chunks settle out before putting
> them in for counts; we almost never see spicules in the hemacytometer
> chambers. I hope clogging won't be a problem with the counter, but
> that's one of the things I'd hoped a manual would settle.
>
> And yes, these are human specimens. I did grow up a blood from my
> horse for chromosomes once, and have done a few mouse and rat samples
> thru the years... but these are just ordinary 2-legged animals :)
>
> So... to get back to the point... the orginal actual question...
>
> Does anyone out there have a CBC5 *with* a manual, who might be
> willing to copy or fax it to us?
>
> Thanks
> Anita
>

"kuhnfucius" <krk4567@comcast . net > wrote in message
news:Bu6dnU1MhNjFwVPbnZ2dnUVZ_g-dnZ2d@comcast . com ...
> Not a dead horse we can learn a lttle about karyotyping. If some of the
> questions are blunt, it is just that I am probing to collect information.
> I always wanted to get into cell cultures and bio-reactors (oh
> well...there is still power ball).
> I know what is charged ($) for FISH and karyotyping. It is expensive.
> Which brings up question why your facility is cheaping out on a good cell
> counter (note I do not state expensive). MDS is probably the most common
> diagnosis on chromosome analysis request. Most are going to be greatly
> diluted with blood (non-diagnostic). How do you know you have enough
> blast cells (e.g. CD-34) for culture (hit or miss?). Do any proficiency
> testing for your hemacytometer use?

I suppose this whole thread begs the question of how many labs are still
using outdated equipment with no backup from suppliers, no calibrators,
etc......

I suspect there's quite a few.