Peters4_Protocol
Project
protocol
—
Contents
Workflow
and
sampling
Equipment
Reagents,
supplies,
and
solutions
Procedure
for
CBC
with
differentials
using
ADVIA
system
Data
References
Workflow
and
sampling
Workflow
Test |
Procedure
performed |
Volume
(µL)
|
Data
collected |
1 |
Body
weight
measurement
|
- |
bw |
2 |
Blood
collection;
samples
are
kept
at
24°C
(for
<3
h)
until
testing
|
270 |
- |
3 |
Hematology
samples
are
run
on
the
ADVIA
120
Hematology
Analyzer |
210 |
Complete
blood
cell
counts
(CBC)
with
differential
|
Equipment
Bayer
ADVIA
120
Hematology
Analyzer
(Bayer
Diagnostic
Division
(Siemens),
Tarrytown,
NY,
USA.
More
equipment
information
ADVIA®
120
Hematology
System
Product
Features
Panel
A
shows
the
Bayer
ADVIA
120
hematology
analyzer
system.
Panel
B
shows
the
internal
layout
of
the
ADVIA,
and
the
various
reagents
used
in
the
system.
Panel
C
shows
the
hematology
setup,
equipped
with
a
computer
and
a
printer.
Panel
D
shows
a
typical
computer
screen-output.
Reagents,
supplies,
solutions
- Microhematocrit
tubes:
heparinized,
75
mm,
75
µL
volume
size
(Fisherbrand®)
- Blood
collection
tubes:
Microtainer®
tube
containing
lyophilized
2EDTA
(BD
(Becton-Dickinson)
Diagnostics,
Franklin
Lakes,
NJ)
or
(Fisher
Catalogue
#
36-5974)
- Toothpicks
- Acclimation
to
test
conditions
- In
general
all
mice
are
brought
into
the
procedure
room
and
are
tested
within
1h.
Procedure:
CBC
with
differential
and
reticulocyte
count
using
Bayer
ADVIA
120
Hematology
Analyzer
I.
Standard
blood
collection
a.
Mice
at
6-7.5,
12-13.5,
and
18-19.5
months
of
age
are
fasted
for
4
hrs
(7:30
to
11:30
am)
before
they
are
bled.
b.
Body
weight
measurements
are
recorded
just
before
blood
samples
are
drawn.
c.
270
µL
of
blood
sample
(about
11
drops
plus
one
capillary
tube)
is
collected
via
a
retro-orbital
bleed
using
a
heparin-coated
capillary
tube
directly
into
BD
Microtainer
tube
coated
with
lyophilized
K2EDTA.
d.
The
freshly
drawn
blood
is
quickly
and
gently
mixed
with
the
anticoagulant
using
a
toothpick.
Vortexing
or
inverting
the
sample
is
not
done
in
order
to
preserve
the
proper
ratio
of
anticoagulant
to
blood
and
also
to
preserve
the
integrity
of
the
Red
Blood
Cells.
e.
Whole
blood
specimens
are
kept
at
room
temperature
until
analysis,
however,
anticipated
delay
of
greater
than
3
hrs
required
sample
refrigeration,
and
subsequent
return
to
room
temperature
before
testing
II.
Blood
cell
count
with
differential
using
Bayer
ADVIA
120
system
a.
Approximately
210
µL
of
un-clotted
whole
blood
is
transferred
to
a
2nd
microtainer
and
is
analyzed
using
the
ADVIA
120
hematology
system
according
to
manufacturer's
protocol.
b.
For
additional
information
see
The
Aging
Center
at
The
Jackson
Laboratory.
Data
collected
by
investigator
Submitted
measurements:
White
blood
cell
count
(WBC),
red
blood
cell
count
(RBC),
neutrophil
count
(NEUT),
lymphocyte
count
(LYMPH),
monocyte
count
(MONO),
eosinophil
count
(EOS),
basophil
count
(BASO),
large
unstained
cell
(LUC),
and
reticulocytes
(Retic)
count,
%
neutrophil,
%
lymphocytes,
%
monocytes,
%
eosinophils,
%
LUC,
%
basophils,
mean
platelet
volume
(MPV),
and
platelet
(PLT)
count;
red
cell
parameters
including
hematocrit
(HCT),
hemoglobin
(HGB),
mean
RBC
corpuscular
hemoglobin
content
(MCH)
and
concentration
(MCHC),
mean
RBC
corpuscular
volume
(MCV),
%
reticulocytes
(Retic),
and
reticulocyte
corpuscular
hemoglobin
content
(CHr)
for
6,
12,
18,
and
24
months
old
mice.
This
figure
illustrates
a
close-up
display
of
the
computer
screen
result
in
Panel
D
above.
|
|
Test age:
6, 12, 18, 24 mo