Blood Banking Applications Using the

Appli cat i on N ote

Thermo Scientific Sorvall BP 8 and 16 and
Heraeus Cryofuge 8 and 16 Centrifuges
Romana Hinz, Thermo Fisher Scientific, Osterode am Harz, Germany

Key Words Platelets from Whole Blood

Blood processing, Blood bag systems, Blood bank protocols, (Buffy-coat (BC) Method)
Large capacity blood banking centrifuges, HAEMAFlex rotors In European countries, platelets preparation is done by
buffy-coat (BC) method[7].

Introduction
Blood banks collect, process, store and distribute blood
and blood products[1]. After collection, whole blood is
separated into its main components: red blood cells
(RBCs), platelets and plasma, which are used effectively
for patient purpose, while white blood cells are depleted[2].
RBCs transport oxygen to body tissues, platelets help the
blood clot, and plasma has specific proteins that allow
proper regulation of coagulation and healing[3].

A key instrument in the blood banking workflow is a

centrifuge. Centrifuges separate whole blood into RBCs, Figure 1: Whole blood processing with the buffy-coat
platelets and plasma. The NEW Thermo Scientific™ (BC) method.
Sorvall™ BP 8 and 16 and Heraeus™ Cryofuge™ 8 and 16
blood banking centrifuges provide enhanced capacity of The first centrifugation step (hard spin) is used initially to
16 x 500 mL blood bags and user-friendly design, separate whole blood into three components: RBCs,
combined with the quick set-up of traceable, reproducible plasma and a BC layer. The components are extracted into
runs for simplified blood processing productivity[4], [5]. a so-called, “top and bottom” collection set, in which
RBCs and plasma are transferred to storage bags and the
This application note introduces the Sorvall BP 8 and 16 BC layer is left in the primary collection bag. This BC
and Heraeus Cryofuge 8 and 16 blood banking layer contains platelets, white blood cells (WBCs), plasma
centrifuges and presents general guidelines to the different and some RBCs. Components can also be extracted into a
protocols for blood component production. This paper top-top blood bag system.
gives an overview of possible methods for the preparation
of blood components and guidance for the correct use of Subsequently, pools of 4-6 ABO-matched BCs are made
centrifuge accessories. A troubleshooting guide for the and either a plasma unit or a platelet additive solution
improvement of blood product yields and instruction on is added.
how to convert protocols from one rotor to another is The second centrifugation step (soft step) is used to
also included. produce PCs which are then extracted with or without
Blood Processing leukofiltration.
Blood component preparation is performed to separate
blood components from whole blood. RBCs and plasma
are produced by a single-step hard spin centrifugation.
Platelet concentrates (PC), RBCs and plasma are prepared
by a two-step centrifugation. The two main procedures of
preparing PCs are either by the platelet-rich plasma (PRP)
method or by buffy-coat (BC) method[6].
2 Platelets from Whole Blood (Platelet-Rich Plasma Thermo Scientific large capacity centrifuges to customize
(PRP) Method) results for maximum yields[4], [5].
Mainly in the United States, platelets are prepared from
When used with the new Thermo Scientific™
whole blood by the PRP method[7].
HAEMAFlex™ 16 blood processing swinging bucket
rotor, the Sorvall BP 16 and Heraeus Cryofuge 16
centrifuges have the ability to run up to 16 blood bag
systems in a single run, each with maximum of 500 mL
collection volume[8].

Rotor Buckets
Rotor buckets are used to hold blood bag liners. Rotor
buckets are also available with
the Thermo Scientific™ Dura-
Coat™ nickel coating, providing
protection against corrosion due
to moisture, chemicals or alkaline
solutions that otherwise can
Figure 2: Whole blood processing with the PRP method.
weaken the structural integrity of
The first centrifugation step (soft spin) results in RBCs a metal rotor bucket.
and PRP. PRP is extracted with or without leukofiltration
Blood Bag Liner
into a so-called, “satellite blood bag” and the RBCs are
The Thermo Scientific blood bag
left in the primary bag.
liner enables easy transportation
The PRP contains platelets, plasma and WBCs. The and stabilization of blood bags
secondary hard spin centrifugation produces platelet-poor for processing.
plasma (PPP) and a platelet pellet. The PPP is extracted
Liner Stand
into a satellite bag and the platelet pellet is resuspended
The Thermo Scientific liner stand
in plasma.
simplifies the bucket loading and
Red Blood Cells/ Plasma Separation unloading process, and can help
After a hard spin leukoreduced, whole blood is separated improve the quality of the blood separation.
into its two main components: RBCs and plasma[7].
Plasma is extracted into a satellite bag while RBC is left in
the primary bag.

Figure 3: Blood processing with RBC/ plasma separation. Balancing Spacers, Weights and Plates
Thermo Scientific spacers are used to compensate low
volume blood bags. Thermo Scientific weights and plates
are used to counterbalance blood bags with
different weight.

Guidelines for Operating Centrifuge

Accessories
Blood bags should be packed following the instructions of
the blood bag manufacturer [9], [10], [11]. Always observe
blood bag manufacturers’ specified protocols and
recommendations. Before using a new or untested type of
blood bag system, it is recommended to perform on-site
Blood Bank Centrifuge and Accessories validation and optimization to predetermine suitability
The Sorvall BP 8 and 16 and Heraeus Cryofuge 8 and 16 with your particular applications and run parameters.
blood banking centrifuges include innovative features
such as energy-saving windshielded rotors with Thermo After packing, blood bags must be placed into a liner. The
Scientific™ Eco-spin™ technology, the Thermo Scientific™ tubing must be put between the bags with the bag tabs
Centri-Touch™ user interface and an automatic front-to- remaining upright to prevent them from becoming tangled
back centrifuge door opening and closing with the around the rotor body during centrifugation.
Thermo Scientific™ Auto-Door™ function and Thermo
Scientific™ Auto-Lid™ rotor lid opening. Moreover, not
only are 11 acceleration and 12 deceleration profiles
provided, but profiles can also be loaded from legacy
 The relative centrifugal force (RCF) is the ratio of the 3
centrifugal acceleration at a specified radius and speed to
the standard acceleration of gravity. The formula to
calculate relative centrifugal force in “g” is as follows:
(RPM)2
RCF = 1,118r
10002

where r is the radius in millimeters, and RPM stands for

Revolutions per Minute.
Figure 4: Incorrect loading where tubing was not properly secured.
At the beginning of the centrifugation process leukocytes
Blood bags that are not properly loaded can cause rotor and red blood cells (RBCs) sediment more rapidly than
failure which could possibly result in leakage or breakage platelets. Leukocytes and RBCs have a higher volume (see
of blood bag systems. Leakage and/or breakage can cause Table 1). In the middle of the centrifugation process,
contamination. (depending on centrifugation time and speed) leukocytes
Blood bags with a low volume must be compensated by and RBCs settle in the lower half of the bag and the upper
using spacers or balance bags. Otherwise, without half contains platelets surrounded by plasma. Then
compensation, low volume blood bags could result in red platelets sediment down, due to the centrifugal force, and
cell traps. As balancing bags could easily break after the leukocytes, situated in the RBCs mass (fluid of higher
several centrifugation runs, select spacers for use over a density), move up.
longer time period. Table 1: Density and volume of blood components[12].

Mean Density Mean Volume

(g/mL) (10-15 liter)
Plasma 1.026 n/a
Platelets 1.059 9
Monocytes 1.062 470
Lymphocytes 1.070 230
Neutrophils 1.082 450
Figure 5: Incorrect loading of bags without compensation. Red Blood Cells 1.100 87

At the end of the centrifugation process, (depending on

centrifugation time and speed) platelet-poor plasma (PPP)
is in the upper part of the bag and RBCs at the bottom.
Platelets are in the top part of the interface RBCs/ plasma
layer, and the majority of leukocytes are in the low part of
the interface.

Since there is a relationship between the physical proper-

ties of blood and the physical principles of centrifugation
that impact separation, the optimal centrifugation for
blood component production is achieved by determina-
tion of the appropriate centrifuge parameters such as
time and speed, and the optimal acceleration and
deceleration profile.

Centrifugation conditions for blood component

Figure 6: Correct loading. preparation are shown in Tables 2, 3 and 4. These
guidelines are based on technical manuals and were
Optimal Centrifugation Parameters validated in the Sorvall BP 8 and 16 and Heraeus
Blood separation is the partial separation of particles from Cryofuge 8 and 16 blood banking centrifuges[12], [13], [14].
a liquid by gravity through sedimentation. The rate of Table 5 shows a troubleshooting guide to improve blood
sedimentation is a function of liquid viscosity, particle component production. An adjustment in speed by
density and particle size, concentration of the solution 200 rpm increments or time by 30 seconds should be
and the force of gravity. To speed up sedimentation, a done. The protocol must be adjusted until the desired
centrifuge is used. As a rotor spins in a centrifuge a yield of products is obtained.
“centrifugal force“ is applied to each particle in the
sample; this can be many thousand times the force of
gravity (g) [12].
4 Table 2: Centrifuge conditions for whole blood processing with the buffy-coat method using the Sorvall BP 8 and 16 and Heraeus
Cryofuge 8 and 16 centrifuges.
Thermo Scientific Speed Time2 Temperature Acceleration Deceleration
Method1 Spin
Rotor (rpm) (min:sec) (°C) Profile Profile
1st spin: 3744 10:00 22 ± 2 9 4
HAEMAFlex 6
2nd spin: 1382 9:30 22 ± 2 3 2
Platelets 1st spin: 3393 10:00 22 ± 2 9 4
from WBC HAEMAFlex 8
2nd spin: 1294 9:30 22 ± 2 3 2
(Buffy-coat
Method) 1st spin: 3347 10:00 22 ± 2 9 4
HAEMAFlex 12
2nd spin: 1282 9:30 22 ± 2 3 2
1st spin: 3201 10:00 22 ± 2 9 4
HAEMAFlex 16
2nd spin: 1242 9:30 22 ± 2 3 2
Note: The given values are only a guideline; user should test different values to find optimized centrifuge conditions.
1
500 mL blood bag systems.

Table 3: Centrifuge conditions for whole blood processing with the PRP method using the Sorvall BP 8 and 16 and Heraeus Cryofuge 8
and 16 centrifuges.

Thermo Scientific Speed Temperature Acceleration Deceleration

Method1 Spin ACE
Rotor (rpm) (°C) Profile Profile
1st spin: 3025 1.70E+07 22 ± 2 9 7
HAEMAFlex 6
2nd spin: 3832 5.5 E+07 22 ± 2 9 7
1st spin: 2704 1.70E+07 22 ± 2 9 7
HAEMAFlex 8
Platelets 2nd spin: 3427 5.5 E+07 22 ± 2 9 7
from PRP 1st spin: 2742 1.70E+07 22 ± 2 9 7
HAEMAFlex 12
2nd spin: 3474 5.5 E+07 22 ± 2 9 7
1st spin: 2587 1.70E+07 22 ± 2 9 7
HAEMAFlex 16
2nd spin: 3278 5.5 E+07 22 ± 2 9 7
Note: The given values are only a guideline; user should test different values to find optimized centrifuge conditions.
1
500 mL blood bag systems.

Table 4: Centrifuge conditions for whole blood processing with the PRP method using the Sorvall BP 8 and 16 and Heraeus Cryofuge 8
and 16 centrifuges.
Thermo Scientific Recommended Time2 Temperature Acceleration Deceleration
Method1 Spin
Rotor Speed (rpm) (min:sec) (°C) Profile Profile
HAEMAFlex 6 1st spin: 3744 10:00 22 ± 2 9 4
Red Blood
Cell/ HAEMAFlex 8 1st spin: 3393 10:00 22 ± 2 9 4
Plasma HAEMAFlex 12 1st spin: 3347 10:00 22 ± 2 9 4
Separation
HAEMAFlex 16 1st spin: 3201 10:00 22 ± 2 9 4
Note: The given values are only a guideline; user should test different values to find optimized centrifuge conditions.
1
500 mL blood bag systems.

Table 5: Troubleshooting guide to improve blood product yields.

1st Spin 2nd Spin

Problem/ Observation 1st Spin Action 2nd Spin Action
Finding Finding
Platelet pellet appears firm, well packed OK Keep speed and time as is OK Keep speed and time as is
Platelet concentrate has aggregates present OK Keep speed and time as is Too hard Decrease time or speed
Platelet pellet appears soft, loosely packed OK Keep speed and time as is Too soft Increase time or speed
Plasma and red blood cell volume acceptable OK Keep speed and time as is OK Keep speed and time as is
Plasma volume high, red blood cell volume low Too hard Decrease time or speed OK Keep speed and time as is
Plasma volume low Too soft Increase time or speed OK Keep speed and time as is
Platelet yield and plasma volume acceptable OK Keep speed and time as is OK Keep speed and time as is
Platelet yield is low and pellet appears firm Too hard Decrease time or speed OK Keep speed and time as is
Platelet yield is low and pellet appears soft Too hard Decrease time or speed Too soft Increase time or speed
Platelet yield acceptable and plasma volume low Too soft Increase time or speed OK Keep speed and time as is
No distinct red cell and plasma line: ‘Bloody interface’ Too hard Decrease slow stop rate OK Keep slow stop rate same
Conversion to Sorvall BP8/16 and Heraeus Summary
SPANISH
Cryofuge 8/16 Centrifuges This application has not only introduced the Thermo
As an example,
thermoscientific.es centrifugation conditions of a sample at Scientific Sorvall BP 8 and 16 and Heraeus Cryofuge 8

Appli cat i on N ote

3300 rpm in a Thermo Scientific ™
Sorvall ™
RC 3BP and 16 blood banking centrifuges, but also presented
© 2015 Thermo Fisher Scientific Inc. Reservados todos los derechos. Las marcas comerciales son propiedad de Thermo Fisher Scientific Inc. y sus filiales.
centrifuge
Características, with yaprecios
condiciones Thermo Scientific
están sujetos
™
HBB-6
a posibles cambios. Algunosswinging different
productos no están disponibles protocols
en todos los países. Consulte losfor blood component production using
detalles
con su representante
bucket rotorcomercial
withlocal.
a radius of 258 mm must be adjusted these new centrifuges. It also includes guidance for the
when used
Alemania +49 6184in the Sorvall BP 8 (with
90 6000 HAEMAFlex
Francia +33 2 2803 2180 6 correctRusia use+7 of812
centrifuge
703 42 15 accessories, a troubleshooting
Australia
swinging +61 bucket
39757 4300 rotor, rotor radius Holanda
of 255 +31mm).
76 579 55 55 guide and Suiza an+41instruction
44 454 12 12 of how to convert protocols from
Austria +43 1 801 40 0 India +91 22 6716 2200 UK/Irlanda +44 870 609 9203
Bélgica +32 53 73 42 41 Italia +39 02 95059 552 one rotor to another.
USA/Canadá +1 866 984 3766
Table+800
China 6: Maximum
810 5118 orrotor radius with theJapón
new+81Thermo
3 5826Scientific
1616
+400
blood650banking
5118 centrifuges and rotors. Nueva Zelanda +64 9 980 6700 References
Otros países de Asia +852 2885 4613
España/Portugal +34 93 223 09 18 Paises Nórdicos/Bálticos/CIS Otros paises
[1] American red +49 6184
cross 90 6000 (2016).
[online]
+358 10 329 2200 o +33 2 2803 2180
Thermo Scientific http://www.redcross.org/blood.
Rotor Radius
Centrifuge and Bucket
Max (mm) [2] Higgins, Vl, Leukocyte-reduced blood components:
Rotor
ITALIAN patient benefits and practical applications, May; 23 (4);
Sorvall BP 8 or Single blood bag bucket 255 659 – 67 (1996).
thermoscientific.com
Heraeus Cryofuge 8 [3] Rodney A. Rhoades, David R. Bell Medical Physiology,
© 2015centrifuge
Thermo Fisher with
Scientific Inc. Tutti i diritti riservati. Tutti gli marchi sono di proprietà di Thermo Fisher Scientific Inc. e dellefor
Principles sue filiali.
ClinicalSpecifiche,
Medicine, 4th edition, (2013).
condizioni e tariffe possono
HAEMAFlex 6 and SingleNonblood
subire variazioni. bag bucket
tutti i prodotti sono disponibili in tutti i258
paesi. Per maggiori dettagli consultare il rivenditore locale.
with filter pack [4] Thermo Scientific Sorvall BP 8/16 manual, 16.02.2016.
HAEMAFlex 8 rotor
Australia +61 39757 4300 India +91 22 6716 2200 Svizzera +41 44 454 12 12
Austria +43 1 801 40 0 Italia +39 02 95059 552 [5] Thermo Scientific
UK/Irlanda +44 870Heraeus
609 9203 Cryofuge 8/16 manual,
BelgioSorvall
+32 53BP
73 8
42or
41 Single blood bagOlanda
bucket 297
+31 76 579 55 55 USA/Canada +1 866 984 3766
16.02.2016.
Heraeus
Cina Cryofuge
+800 810 5118 or 8 Nuova Zelanda +64 9 980 6700
+400centrifuge
650 5118 with Europa del Nord/CIS [6] HansAltre Nazioni Asiatiche
Gulliksson, +852 2885
Platelets from4613
platelet-rich-plasma versus
Francia +33 2 2803 2180 Single blood bag+358bucket
10 329 2200 299 Nazioni non in elenco +49 6184 90 6000
buffy-coat-derived platelets: what is the difference?; Rev Bras
HAEMAFlex
Germania 8 rotor
+49 6184 90 6000 Russia +7 812 703 42 15 o +33 2 2803 2180
Giappone +81 3 5826 1616 with filter pack
Spagna/Portogallo +34 93 223 09 18 Hematol Hemoter, 34 (2); 76 – 77 (2012).
Sorvall BP 16 or Double blood bag bucket 291 [7] Christopher D. Hillyer, Blood Banking and Transfusion
GERMAN
Heraeus Cryofuge 16 Medicine: Basic Principles & Practice, 2nd edition, (2007).
centrifuge with
thermoscientific.de [8] Thermo Scientific Sorvall BP 8/16 and Heraeus Cryofuge
HAEMAFlex 12 rotor blood banking centrifuges, brochure, 16.02.2016.
© 2015 Thermo Fisher Scientific Inc. Alle Rechte vorbehalten. Alle Warenzeichen sind Eigentum von Thermo Fisher Scientific Inc. bzw. nationalen
Sorvall BP
Niederlassungen. Änderungen Double blood
16 or von Spezifikationen, bag bucket
Bedingungen 316Nicht alle Produkte sind in allen Ländern erhältlich. Ausführliche
und Preisen vorbehalten.
[9] Fresenius. [Online] (21.01.2016).
Heraeus erhalten
Informationen Cryofuge 16Ihrem Thermo Fisher Scientific Ansprechpartner.
Sie von
https://www.fresenius.de/689.htm.
centrifuge with
Australien +61 39757 4300 Indien +91 22 6716 2200 Russland +7 812
[10] Haemonetics. 703 42 15(21.01.2016).
[Online]
HAEMAFlex 16 rotor
Belgien +32 53 73 42 41 Italien +39 02 95059 552 Spanien/Portugal +34 93 223 09 18
http://wbt.haemonetics.com/en/Products.aspx.
China +800 810 5118 or Japan +81 3 5826 1616 Schweiz +41 44 454 12 12
+400 650 5118 Niederlande +31 76 579 55 55 UK/Irland +44 870 609 9203
[11] Macopharma. [Online] (21.01.2016).
Deutschland nationale kostenlose Hotline Neuseeland +64 9 980 6700 USA/Kanada +1 866 984 3766
The conversion
0800 1 536 376 is done as follows: Nordeuropa/Baltikum/CIS http://www.macopharma.com/category/transfusion/blood-
Deutschland
1) Calculateinternational
and determine RCF for +358 10 329 2200
HBB-6 rotor: processing/blood-bags/.
Andere asiatische Länder +852 2885 4613
+49 6184 90 6000 Österreich +43 1 801 40 0 Andere Länder +49 6184 90 6000
Frankreich +33 2 2803 2180 [12] Guide
oder to
+33the preparation,
2 2803 2180 use and quality assurance of
2) Radius in mm
(RPM)2 blood components, European committee (partial agreement)
RCF = 1,118r = 3146 on blood transfusion, 16th Edition (2010).
FRENCH 10002
[13] AABB. [Online] (21.01.2016).
So, centrifugation of a sample at 3300 rpm in a Sorvall
thermoscientific.fr https://www.aabb.org/tm/coi/Documents/coi1113.pdf.
RC 3BP centrifuge with the HBB-6 swinging bucket rotor
[14] Validation
© 2015 Thermo Fisher Scientific Inc. Tous droits réservés. Les marques déposées sont des marques commerciales
will deliver a centrifugal force of 3146 x g. was Fisher
ou déposées de Thermo doneScientific
at Bonfils
Inc. Blood Center,
et de ses filiales. Les caractéristiques, conditions et tarifs sont susceptibles d’être modifiés. Tous les produits ne717
sont pas disponibles dans
Yosemite tous lesDenver,
Street, pays. Pour tout
CO 80230.
renseignement, veuillez vous adresser à votre distributeur local.
3) Calculate RPM for HAEMAFlex 6 rotor:
Allemagne +49 6184 90 6000 RCF France +33 2 2803 2180 Russie +7 812 703 42 15
RPM =
Australie +61 39757 4300 1000 2
Inde +91 3320
= 22 6716 2200 Suisse +41 44 454 12 12
Autriche +43 1 801 40 0
1,118r Italie +39 02 95059 552 UK/Irlande +44 870 609 9203
Belgique +32 53 73 42 41
Centrifugation of the sample in theJapon
Chine +800 810 5118 or
+81 3 5826 1616
Sorvall BP 8 with the
Nouvelle-Zélande +64 9 980 6700
USA/Canada +1 866 984 3766

HAEMAFlex
+400 650 5118 6 swinging bucket rotor needs
Pays Bas to579
+31 76 be 55
done
55 at Autres pays d’Asie +852 2885 4613
Espagne/Portugal +34 93 223 09 18 Pays Nordiques/Baltes/CEI Autres Pays +49 6184 90 6000
3320 rpm. +358 10 329 2200 ou +33 2 2803 2180

thermofisher.com/bloodbankcentrifuges
thermoscientific.com
© 2016 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries unless otherwise
© 2015 Thermo
indicated. Heraeus Fisher
is aScientific
registered Inc.trademark
All rights reserved. All trademarks
of Heraeus are thelicensed
Holding GmbH propertytoofThermo
Thermo Fisher
Fisher Scientific
Scientific.Inc.Specifications,
and its subsidiaries.
termsSpecifications,
and pricing are subject to
terms and
change. Notpricing are subject
all products are toavailable
change. in Notallallcountries.
products are available
Please in allyour
consult countries. Pleaserepresentative
local sales consult your local
for sales representative for details.
details.

Australia +61 39757 4300 India toll free 1800 22 8374 Russia +7 812 703 42 15
Austria +43 1 801 40 0 India +91 22 6716 2200 Spain/Portugal +34 93 223 09 18
Belgium +32 53 73 42 41 Italy +39 02 95059 552 Switzerland +41 44 454 12 12
China +800 810 5118 or Japan +81 3 5826 1616 UK/Ireland +44 870 609 9203
+400 650 5118 Netherlands +31 76 579 55 55 USA/Canada +1 866 984 3766
France +33 2 2803 2180 New Zealand +64 9 980 6700
Germany national toll free 0800 1 536 376 Nordic/Baltic/CIS countries Other Asian countries +852 2885 4613
Germany international +49 6184 90 6000 +358 10 329 2200 Countries not listed +49 6184 90 6000
ANCFGBLOODBANK 0216