Immune Responsiveness and Outcome After Aortic Valve Surgery.

Barts Heart Centre is a 255-bedded specialist cardiac centre where approximately 200 primary isolated open aortic valve replacement procedures are performed per year. It is closely associated both geographically and academically with the William Harvey Research Institute (WHRI), which forms part of Queen Mary University, London (QMUL). This will allow laboratory analysis of all collected blood samples within 15 minutes if necessary. Barts Heart Centre is one of the largest cardiac units in Europe and a key research objective of QMUL/WHRI is to support Centrifuge tube cardiac research.

Sample collection and storage.

Following written informed consent all patients will be pre-operatively risk-scored by means of ASA, Euroscore 2 and conventionally risk-scored for the development of surgical site infection.

• Blood (44ml total) will be taken from 150 aortic valve replacement patients immediately prior to surgery (T0).
• Blood (7ml) will be collected into a PAXgene tube and stored at -80° for later genetic analysis. A clotted sample (7ml) will be collected in a standard gold top SST tube, left to clot then centrifuged for 10 mins at 1000g, the serum separated and stored at -80° for later analysis of antibody levels.
• In addition, 30 ml blood will be taken for study of peripheral blood mononuclear cells (PBMCs), drawn into standard purple top EDTA tubes (anticoagulant). These will be separated within 3 hours of collection by Ficoll-Paque density gradient centrifugation, whereby PBS-diluted blood is carefully layered over Ficoll and spun brake off in a centrifuge at 400g for 30 minutes.
• The PBMC layer is then extracted and washed twice in PBS and stored in 10% dimethyl sulfoxide freezing solution in liquid nitrogen so that they can be thawed, and batch analysed at a later date.

Sample analysis

PBMCs will be counted using a haemocytometer, then cultured in RPMI and 10% serum with either LPS, α-toxin or unstimulated for 24 hours, after which they will be analysed immediately as detailed below.

• Soluble Mediators (Plasma)

– Storage: Ethylenediaminetetraacetic acid (EDTA, BD Biosciences Vacutainer, 9mL) anticoagulated blood will be centrifuged (1200g, 10mins, 20°C) and plasma stored at -80°C within 2hrs of collection

– Analysis: Cytokines: Meso Scale Discovery (MSD) V-PLEX Proinflammatory Panel 1 will be employed to quantify IFN-γ, IL-10, IL-12p70, IL-13, IL-1β, IL-2, IL-4, IL-6, IL-8, and TNF-α. Plates will be read using a MSD QuickPlex SQ 120 imager (Wiliam Harvey Institute, QMUL).

Antibodies: These will be measured by means of ELISA assays against staphylococcal antigens (α-toxin, teichoic acid) and core moieties of the endotoxin molecule (EndoCAb).

– Whole blood LPS/α-toxin Stimulated Cytokine Release

– Technique: As previously described and validated in our laboratory, heparinized blood will be stimulated for 4hrs (37°C, 250rpm) with 1ng/ml LPS within 2hrs of draw. After incubation, samples will be centrifuged (1200g, 10mins, 20°C) and supernatant stored at -80°C.

– Cytokine quantification: As per ‘soluble mediators’ MSD V-PLEX Proinflammatory Panel 1 will be employed to quantify TNF-α (primary outcome as metric of immune competence) and other cytokines. Cytokines will be expressed as a factor of the number of circulating monocytes.

Flow Cytometric Immunophenotyping and Functional Assessment of Leukocytes

Flow Cytometry: All samples will be analysed on the same machine (Becton Dickinson (BD) Fortessa, Rayne Building, UCL). Standardisation and comparison of output will be achieved via employment of constant voltages and compensation matrix throughout with daily matching of values to a single batch of Cytometry Setup and Tracking (CS&T) beads (BD). Leukocyte cell surface staining will be conducted using antibodies principally supplied by BD. Staining, data capture and storage will be conducted in accordance with a single study standard operating procedure. 5 panels (up to 14-colour) and two functional assays will be performed at each time-point. Data will be analysed in FlowJo version 10.5.

Surface Marker Staining:

Quantification and calibration panel: CD45, CD56, CD3, CD4, CD8, CD19, CD14, CD16, performed in a BD TruCount tube to enable enumeration of cell subsets Cell specific panels (incorporating markers of sub-categorisation and markers of immune competence)

– Monocyte/Myeloid: CD3, CD19, CD56, CD163, CD16, CD33, CD141, CD206, CD274, CD14, CD1c, CD11c, HLA-DR, CD80, CD123, CD83

– Neutrophil: CD3, CD19, CD56, HLA-DR, CD64, CD62L, CD11b, CD88, CD66b, CD14, CD11c, CD16, CD184, CD182

– Lymphocyte: CD19, CCR6, CD45RA, CD3, CD4, CD8, CXCR3, CD127, CD62L, CD25, CD56, CD27, CD279 HLA-DR Expression: Will be quantified on CD14+ monocytes using BD QuantiBrite beads.

Functional Measures Phagocytosis: The ability of neutrophils and monocytes to phagocytose opsonised FITC-labelled E.coli bacteria will be quantified via the PhagoTest assay (BD) as per manufacturer’s instructions Reactive oxygen ‘burst’: Quantitative determination of leukocyte oxidative burst will be undertaken using the PhagoBurst assay (BD) in response to un-labelled opsonized E.coli, phorbol 12-myristate 13-acetate (PMA) and the chemotactic peptide N-formyl-Met-Leu-Phe (fMLP).

The pre-surgery sample PBMCs that will be stimulated with LPS (type and concentration to be optimized via dose response curve on healthy volunteers) and α-toxin will undergo flow cytometry to measure TLR4 and TLR5 surface expression, HLA-DR surface expression and quantification of NF-kB using appropriate antibodies validated in the literature.

At the end of stimulation, cells will be harvested, and RNA extracted for quantitative (q)PCR to evaluate AID mRNA expression. Although B cells in the PBMC cultures have been stimulated in the presence of other cell types, primarily T cells and monocytes-macrophages, our endpoint is to measure a B-cell response, as AID is exclusively expressed in B cells.

The Impact of Regional Anaesthesia on Hormone Levels in Thoracic Surgery.

• Basic aspects of thoracic anaesthesia are general anesthesia often combined with regional anesthesia, intubation with double lumen tube and separation of lung ventilation. Proper assessment of pain and adequate analgesia in intraoperative and postoperative period is a challenging issue for medical practitioners.
• Intraoperative trauma may lead to many metabolic implications and disturbance of haemostasis, what can be reflected in change of blood and saliva hormone and other substance levels, such as alpha-amylase, cortisol, testosterone, secretory IgA, β-endorphin, nerve growth factor, calcitonin gene-related protein and P substance.
• The aim of this study is to assess the impact of regional anesthesia on hormone levels in postoperative period. Saliva was collected from participants in order to perform laboratory tests, using a special disposable Salivette tube (Sarstedt AG & Co, Germany). Saliva was collected by placing a sterile tampon under the tongue or chewing for 30-45 seconds.
• The soaked saliva pad was then placed in a suspended insert with a perforated bottom. The insert with a tampon was placed in a centrifuge tube and closed with a stopper. Next the tube was centrifuged (1000 x g for 10 min.) to obtain a ready to test saliva supernatant. Approximately 0,7 ml of the supernatant from every sample collected was used for further testing. Samples were frozen after centrifugation at – 85°C until performing laboratory tests. Blood was collected for laboratory tests from the ulnar vein. Blood for testing was collected using disposable equipment in a volume of 5ml into a tube containing ethylenediaminetetraacetic acid (EDTA) and aprotinin.

15 ml & 50 ml Centrifuge Tubes
TCF001-00-15mICentrifugeTubes450mlCentrifugeTubes4	Vazyme	15 mI Centrifuge Tubes*450 ml Centrifuge Tubes*4
Centrifuge Tubes, 50 ml ,PP,Sterile,Plug-Seal Cap, in Bag
LC8005-500	GenDepot	500/CS
Centrifuge Tubes, 15 ml ,PP,Sterile,Plug-Seal Cap, in Bag
LC8003-500	GenDepot	500/CS
Centrifuge Tubes, 50 ml , Black ,PP,Sterile
LC8008-025	GenDepot	25/Bag
Centrifuge Tubes, 50 ml , Black ,PP,Sterile
LC8008-500	GenDepot	500/CS
Centrifuge Tubes, 50 ml , Amber ,PP,Sterile
LC8050-500	GenDepot	500/CS
Centrifuge Tubes, 15 ml , Black ,PP,Sterile
LC8007-050	GenDepot	50/Bag
Centrifuge Tubes, 15 ml , Black ,PP,Sterile
LC8007-500	GenDepot	500/CS
Centrifuge Tubes, 15 ml , Amber ,PP,Sterile
LC8015-500	GenDepot	500/CS
Centrifuge Tubes, 50 ml ,PP,Sterile,Plug-Seal Cap,Self Standing, in Bag
LC8006-500	GenDepot	500/CS
Centrifuge Tubes 0.65 ml Blue 10 bags of 1000
MU11060	Westburg	each
Centrifuge Tubes 0.65 ml Pink 10 bags of 1000
MU11120	Westburg	each
Centrifuge Tubes 1.7 ml Blue 10 bags of 500
MU11430	Westburg	each
Centrifuge Tubes 1.7 ml Pink 10 bags of 500
MU11490	Westburg	each
Centrifuge Tubes 2.0 ml Blue 10 bags of 400
MU11920	Westburg	each
Centrifuge Tubes 2.0 ml Pink 10 bags of 400
MU11980	Westburg	each
Rack S for centrifuge tubes 18 x 15 ml / 10 x 50 ml (45)
TP99017	Westburg	each
Rack L for centrifuge tubes 30 x 15 ml / 20 x 50 ml (30)
TP99019	Westburg	each
Centrifuge Tubes 0.65 ml Green 10 bags of 1000
MU11080	Westburg	each
Centrifuge Tubes 1.7 ml Green 10 bags of 500
MU11450	Westburg	each
Centrifuge Tubes 2.0 ml Green 10 bags of 400
MU11940	Westburg	each
Centrifuge Tubes 0.65 ml Purple 10 bags of 1000
MU11170	Westburg	each
Centrifuge Tubes 0.65 ml Yellow 10 bags of 1000
MU11190	Westburg	each
Centrifuge Tubes 1.7 ml Yellow 10 bags of 500
MU11560	Westburg	each
Centrifuge Tubes 2.0 ml Purple 10 bags of 400
MU12020	Westburg	each
Centrifuge Tubes 2.0 ml Yellow 10 bags of 400
MU12040	Westburg	each
50 mI Centrifuge Tubes
TCF00150-500Case	Vazyme	500 / Case
Centrifuge Tubes 0.65 ml Natural 10 bags of 1000
MU11150	Westburg	each
Centrifuge Tubes 0.65 ml Rainbow 10 bags of 1000
MU11210	Westburg	each
Centrifuge Tubes 1.7 ml Natural 10 bags of 500
MU11510	Westburg	each
Centrifuge Tubes 1.7 ml Rainbow 10 bags of 500
MU11590	Westburg	each
Centrifuge Tubes 2.0 ml Natural 10 bags of 400
MU12000	Westburg	each
Centrifuge Tubes 2.0 ml Rainbow 10 bags of 400
MU12060	Westburg	each
Centrifuge Tubes 0.65 ml Natural 20 bags of 1000
MU17030	Westburg	each
Centrifuge Tubes 1.7 ml Natural 10 bags of 1000
MU17040	Westburg	each
Centrifuge Tubes 2.0 ml Natural 10 bags of 1000
MU17050	Westburg	each
15 mI Centrifuge Tubes
TCF00115-500Case	Vazyme	500 / Case
Centrifuge tubes, glas, with round bottom, 50 ml
CD04-R50G	Phoenix instrument	each
15 mL Centrifuge Tube, Tubes and Caps Packed Separately, Non-Sterile, 50/pk, 500
601151	NEST Biotechnology	500 pcs/cs
Centrifuge tubes, plastic, with round bottom, 50 ml
CD04-R50P	Phoenix instrument	each
50ml Conical Centrifuge Tubes, Green Cap, 25/Bag
CT788-G	Bio Basic	1PK, 25UNIT
50ml Conical Centrifuge Tubes, Yellow Cap, 25/Bag
CT788-Y	Bio Basic	1PK, 25UNIT
Block, 15 x1.5ml centrifuge tubes
BSH100-15	Benchmark Scientific	1 PC
Block, 15 x1.5ml centrifuge tubes
BCM1405-B15	Bio Basic	1 pcs, 1 UNIT
Small centrifuge, 6 x 50 ml, 4000 rpm, incl. rotor for 6 x 50 ml or 6 x 15 ml Tubes
CD-0412-50	Phoenix instrument	each
Rack filled with 20 centrifuge tubes 50 ml, conical (10)
TP91550	Westburg	each
Stand For Centrifuge Tubes For 1.5 ml,
PW102-2NO	EWC Diagnostics	1 unit
Rack filled with 30 centrifuge tubes 15 ml, conical (10)
TP91515	Westburg	each
Centrifuge Tubes 0.65 ml, low binding 10x500
MU11300	Westburg	each
Centrifuge Tubes, 50 ml ,PP,Sterile,Plug-Seal Cap, in Rack
LC8004-500	GenDepot	500/CS
Centrifuge Tubes 2.0 ml Natural 12 x 800
MU1220	Westburg	each
Corning LSE Single Block 5 x 50 mL Centrifuge Tubes - EACH
480129	Scientific Laboratory Supplies	EACH
Dolphin Centrifuge Tubes 2.0 ml Natural 5x1000
MU16140	Westburg	each
29 mm Test Tube Foam Rack (for 50 mL centrifuge tubes) - EACH
MIX1118	Scientific Laboratory Supplies	EACH
Centrifuge Tubes, 15 ml ,PP,Sterile,Plug-Seal Cap, in Rack
LC8002-500	GenDepot	500/CS
16 mm Test Tube Foam Rack (for 15 mL centrifuge tubes) - EACH
MIX1098	Scientific Laboratory Supplies	EACH
Centrifuge Tubes 0.65 ml Natural Low binding 2x500
MU11330	Westburg	each
Centrifuge Tubes 2.0 ml Natural, low binding 4x500
MU12160	Westburg	each
Centrifuge Tubes 2.0 ml Natural Low binding 2x500
MU12180	Westburg	each
Centrifuge Tubes 1.7 ml Natural, Low binding 5x500
MU39640T	Westburg	each
15 mL Centrifuge Tube Rack, 1/pk, 50/cs
610001	NEST Biotechnology	50 pcs/cs
Centrifuge tube 50 ml, conical / triple bagged (50)
TP391050	Westburg	each
Centrifuge Tubes 1.7 ml Natural Lubricated 2 x 500
MU11720	Westburg	each
4-Ways Microtube and Centrifuge Tubes Racks, Red
R023-R	Bio Basic	1 UNIT
Greiner centrifuge tubes - PK300
T2193-300EA	Scientific Laboratory Supplies	PK300
4-Ways Microtube and Centrifuge Tubes Racks, Blue
R023-B	Bio Basic	1 UNIT
Greiner centrifuge tubes - PK1000
T1943-1000EA	Scientific Laboratory Supplies	PK1000
4-Ways Microtube and Centrifuge Tubes Racks, Yellow
R023-Y	Bio Basic	1 UNIT
Centrifuge tube 50 ml, round (180)
TP91056	Westburg	each
4-Ways Microtube and Centrifuge Tubes Racks, Natural
R023-N	Bio Basic	1 UNIT
Block, 15 x 1.5ml or 2.0ml centrifuge tubes (conical)
BSH100-1520	Benchmark Scientific	1 PC
Block, 15 x 1.5ml or 2.0ml centrifuge tubes (conical)
BCM1405-B1520	Bio Basic	1 pcs, 1 UNIT
Centrifuge Tubes, Autoclavable BLUE
CG264-1X100NO	EWC Diagnostics	1 unit
Centrifuge tube 50 ml, conical (360)
TP91050	Westburg	each
Centrifuge tube 50 ml, conical (160)
TP91054	Westburg	each
Centrifuge Tubes, Autoclavable Round
CG276-1X100NO	EWC Diagnostics	1 unit
Centrifuge Tubes, Autoclavable Round
CG277-1X100NO	EWC Diagnostics	1 unit
Centrifuge Tubes, Autoclavable Round
CG278-1X100NO	EWC Diagnostics	1 unit
Centrifuge Tubes, Autoclavable Round
CG279-1X100NO	EWC Diagnostics	1 unit
Block for 5ml centrifuge tubes
H5000-5MT	Benchmark Scientific	1 PC
15 mL Centrifuge Tube, Bulk, Sterile, 50/pk, 500/cs
601052	NEST Biotechnology	500 pcs/cs
Centrifuge tube 15 ml, conical (360)
TP91014	Westburg	each
Centrifuge tube 15 ml, conical (800)
TP91015	Westburg	each
Centrifuge tube 15 ml, conical (800)
TP91115	Westburg	each
Centrifuge Tubes, Autoclavable Conical
CG274-100X1NO	EWC Diagnostics	1 unit
15mL Sterile Centrifuge Tubes PP
ARG1818	Scientific Laboratory Supplies	PK500
50mL Sterile Centrifuge Tubes PP
ARG1824	Scientific Laboratory Supplies	PK500
Block, 20x10mm test tubes or 20x2.0ml centrifuge tubes
BSW10	Benchmark Scientific	1 PC
Block, 20x10mm test tubes or 20x2.0ml centrifuge tubes
BCM1412-BSW10	Bio Basic	1 pcs, 1 UNIT
Eppendorf Adapter for 0.5 mL micro- centrifuge tubes + 0.6 ml Microtainer set of 6 - PK6
E5425716001	Scientific Laboratory Supplies	PK6
Centrifuge tube 15 ml, conical / triple bagged (100)
TP391015	Westburg	each
CapLock 0.5mL Centrifuge Tubes - PK500
ARG1782	Scientific Laboratory Supplies	PK500
Oakridge Centrifuge Tubes 30ml - PK10
31390030	Scientific Laboratory Supplies	PK10
CapLock 1.5mL Centrifuge Tubes - PK1000
ARG1808	Scientific Laboratory Supplies	PK1000
50 mL Centrifuge Tube Rack, 1/pk, 50/cs
610101	NEST Biotechnology	50 pcs/cs
Block, 24 x 0.5ml centrifuge tubes
BSH100-05	Benchmark Scientific	1 PC
Block, 24 x 0.5ml centrifuge tubes
BSW05	Benchmark Scientific	1 PC
Block, 24 x 0.5ml centrifuge tubes
BCM1405-B05	Bio Basic	1 pcs, 1 UNIT
Block, 24 x 0.5ml centrifuge tubes
BCM1412-BSW05	Bio Basic	1 pcs, 1 UNIT
Centrifuge Tubes S 2420 30ml Qfit - PK10
QBC24C24R0	Scientific Laboratory Supplies	PK10
Centrifuge Tubes, Autoclavable RED Cap,
CG272-1X100NO	EWC Diagnostics	1 unit
Block, 4 x 15ml centrifuge tubes
BSH100-150	Benchmark Scientific	1 PC
Block, 2 x 50ml centrifuge tubes
BSH100-500	Benchmark Scientific	1 PC
Block, 12 x 15ml centrifuge tubes
BSW15	Benchmark Scientific	1 PC
Block, 5 x 50ml centrifuge tubes
BSW50	Benchmark Scientific	1 PC
Block, 4 x 15ml centrifuge tubes
BCM1405-B150	Bio Basic	1 pcs, 1 UNIT
Block, 2 x 50ml centrifuge tubes
BCM1405-B500	Bio Basic	1 pcs, 1 UNIT
Block, 12 x 15ml centrifuge tubes
BCM1412-BSW15	Bio Basic	1 pcs, 1 UNIT
Block, 5 x 50ml centrifuge tubes
BCM1412-BSW50	Bio Basic	1 pcs, 1 UNIT
Centrifuge Tubes, with ring (leak proof)
PW1207-1X100NO	EWC Diagnostics	1 unit
Centrifuge Tubes, Autoclavable With RED
CG271-1X100NO	EWC Diagnostics	1 unit
Block for 5mL centrifuge tubes - EACH
MIX7218	Scientific Laboratory Supplies	EACH
Block, 24 x1.5ml centrifuge tubes (conical)
BSW1500	Benchmark Scientific	1 PC
Block, 24 x1.5ml centrifuge tubes (conical)
BCM1412-BSW1500	Bio Basic	1 pcs, 1 UNIT
15mL Sterile Centrifuge Tubes PP - PK500
ARG1814	Scientific Laboratory Supplies	PK500
50mL Sterile Centrifuge Tubes PP - PK500
ARG1820	Scientific Laboratory Supplies	PK500
Falcon Conical 50ml PP Centrifuge Tubes Bagged - PK500
352070	Scientific Laboratory Supplies	PK500
Falcon Conical 15ml PP Centrifuge Tubes Bagged - PK500
352096	Scientific Laboratory Supplies	PK500
15ml Conical Centrifuge Tubes, Polypropylene
28-101	Genesee Scientific	20 Racks of 25 Tubes, 500/Unit
15ml Conical Centrifuge Tubes, Polypropylene
28-103	Genesee Scientific	20 Bags of 25 Tubes, 500/Unit
50ml Conical Centrifuge Tubes, Polypropylene
28-106	Genesee Scientific	20 Racks of 25 Tubes, 500/Unit
50ml Conical Centrifuge Tubes, Polypropylene
28-108	Genesee Scientific	20 Bags of 25 Tubes, 500/Unit
50ml Conical Centrifuge Tubes, Polypropylene
21-106	Genesee Scientific	20 Racks of 25 Tubes, 500/Unit
50ml Conical Centrifuge Tubes, Polypropylene
21-108	Genesee Scientific	20 Bags of 25 Tubes, 500/Unit
50ml Plug-Seal Centrifuge Tubes with Rack, 25 tubes/rack
10-0501	IHC World	-
Kimas high strength centrifuge tubes - PK6
Z514861-6EA	Scientific Laboratory Supplies	PK6
5 mL Centrifuge Tubes, Clear, Round, Lock Cap, Sterile, 200/pk, 2000/cs
603111	NEST Biotechnology	2000 pcs/cs
Centrifuge 5804 + Rotor S-4-72 + adapters 15/50ml tubes - EACH
E5804000565	Scientific Laboratory Supplies	EACH
Tubes Centrifuge 15ml Conical - PK500
CEN9202	Scientific Laboratory Supplies	PK500
CENTRIFUGE TUBE 50 ML 30 X 115 MM - PK500
T2318-500EA	Scientific Laboratory Supplies	PK500
15 mL Centrifuge Tube, Rack(PS Rack), Sterile, 50/pk, 500/cs
601051	NEST Biotechnology	500 pcs/cs
Centrifuge 5804R + Rotor S-4-72 + adapters 15/50ml tubes - EACH
E5805000665	Scientific Laboratory Supplies	EACH
Centrifuge 5810 + Rotor S-4-104 + adapters 15/50ml tubes - EACH
E5810000560	Scientific Laboratory Supplies	EACH
250 mL PP Centrifuge Tubes with Plug Seal Cap, Sterile, 6/pk, 102/cs
622001	NEST Biotechnology	102 pcs/cs
500 mL PP Centrifuge Tubes with Plug Seal Cap, Sterile, 6/pk, 36/cs
623001	NEST Biotechnology	36 pcs/cs
Centrifuge 5810R + Rotor S-4-104 + adapters 15/50ml tubes - EACH
E5811000660	Scientific Laboratory Supplies	EACH
SINGLE BLOCK, 12 X 15ML CENTRIFUGE TUBES
480124	CORNING	1/pk
Centrifuge tube 50 ml, conical, with rim (320)
TP91051	Westburg	each
Block, 24 x 1.5ml or 2.0ml centrifuge tubes
BSW1520	Benchmark Scientific	1 PC
Block, 24 x 1.5ml or 2.0ml centrifuge tubes
BCM1412-BSW1520	Bio Basic	1 pcs, 1 UNIT
250mL centrifuge tube PP (59.8 X 160mm), with screw-cap, 14 bags of 5 tubes
C1250	MTC Bio	70
Block, 15 x 1.5ml Centrifuge Tubes for Dry Bath with Heated Lid
BSH100-15HL	Benchmark Scientific	1 PC
Block, 15 x 1.5ml Centrifuge Tubes for Dry Bath with Heated Lid
BCM1405-B15HL	Bio Basic	1 pcs, 1 UNIT
Falcon Conical 225ml PP Centrifuge Tubes - PK48
352075	Scientific Laboratory Supplies	PK48
5 mL Centrifuge Tubes, Clear, Round, Lock Cap, Non-Sterile, 200/pk, 2000/cs
603011	NEST Biotechnology	2000 pcs/cs
15mL and 50ml Universal Rack for Conical Bottom Centrifuge Tubes - PK20
4366	Scientific Laboratory Supplies	PK20
Falcon Conical 15ml Ps Centrifuge Tubes - PK500
352099	Scientific Laboratory Supplies	PK500
5-Well Centrifuge Tube Box (1.5ml To 2ml Tubes)
R005-G	Bio Basic	1 UNIT
High Speed Centrifuge, Adapt for 1.5/2.0 mL Microcentrifuge Tubes, 500-1200 RPM
102101	NEST Biotechnology	1 pcs/cs
250 mL PP Centrifuge Tubes with Plug Seal Cap, Racked, Sterile, 6/pk, 24/cs
622002	NEST Biotechnology	24 pcs/cs
500 mL PP Centrifuge Tubes with Plug Seal Cap, Racked, Sterile, 6/pk, 24/cs
623002	NEST Biotechnology	24 pcs/cs
Block, 6 x 5ml centrifuge tubes (17mm diameter)
BSH100-5MT	Benchmark Scientific	1 PC
Block, 12 x 5.0ml centrifuge tubes (17mm diameter)
BSW5MT	Benchmark Scientific	1 PC
Block, 6 x 5ml centrifuge tubes (17mm diameter)
BCM1405-B5MT	Bio Basic	1 pcs, 1 UNIT
Block, 12 x 5.0ml centrifuge tubes (17mm diameter)
BCM1412-BSW5MT	Bio Basic	1 pcs, 1 UNIT
Block, 8x5ml centrifuge tubes, for MultiTherm Touch
H5100-5MT	Benchmark Scientific	1 each
Block 15 x 1.5mL Centrifuge Tubes for Dry Bath with Heated Lid - EACH
BLO1254	Scientific Laboratory Supplies	EACH
Centrifuge Tube, 5ml, 200 per. pk. (4 bags of 50)
C1005-T5-Clear	Benchmark Scientific	1 each
Centrifuge Tube, 5ml, 200 per. pk. (4 bags of 50)
C1005-T5_Clear	Benchmark Scientific	each
Centrifuge 5804 + Rotor S-4-72 + adapters 15/50ml tubes- IVD Only - EACH
E5804000560	Scientific Laboratory Supplies	EACH
Centrifuge 5804R + Rotor S-4-72 + adapters 15/50ml tubes- IVD Only - EACH
E5805000661	Scientific Laboratory Supplies	EACH
Centrifuge 5810 + Rotor S-4-104 + adapters 15/50ml tubes- IVD Only - EACH
E5810000564	Scientific Laboratory Supplies	EACH
15ml Centrifuge Tube, Screw Cap, Natural, 100/Bag
BT888-N	Bio Basic	1PK, 100UNIT
Centrifuge 5810R + Rotor S-4-104 + adapters 15/50ml tubes- IVD Only - EACH
E5811000665	Scientific Laboratory Supplies	EACH
50 mL Centrifuge Tube, Bulk, Sterile, 25/pk, 500/cs
602052	NEST Biotechnology	500 pcs/cs
50mL Rack for Conical Bottom Centrifuge Tubes - PK20
4365	Scientific Laboratory Supplies	PK20
15mL Rack for Conical Bottom Centrifuge Tubes - PK20
431355	Scientific Laboratory Supplies	PK20
Falcon Conical 175ml PP Centrifuge Tubes Racked - PK48
352076	Scientific Laboratory Supplies	PK48
Cool rack 9 wells for 15ml centrifuge tubes - EACH
HOM2740	Scientific Laboratory Supplies	EACH
Cool rack 4 wells for 50ml centrifuge tubes - EACH
HOM2742	Scientific Laboratory Supplies	EACH
Falcon Conical 15ml PP Centrifuge Tubes Racked - PK500
352097	Scientific Laboratory Supplies	PK500
Falcon Conical 50ml PP Centrifuge Tubes Racked - PK500
352098	Scientific Laboratory Supplies	PK500
Kimax Reuseable Glass Centrifuge tubes oil 203mm - PK12
TUB2716	Scientific Laboratory Supplies	PK12
15 mL Centrifuge Tube, Amber Color, Rack(PS Rack), Sterile, 50/pk, 500/cs
601201	NEST Biotechnology	500 pcs/cs
50ml Skirted Centrifuge Tube, Green Cap, 25/Bag
TK943-G	Bio Basic	1PK, 25UNIT
Block 6 x 5mL centrifuge tubes (17mm diameter) - EACH
BLO1256	Scientific Laboratory Supplies	EACH
Block 12 x 5.0mL centrifuge tubes (17mm diameter) - EACH
BLO1276	Scientific Laboratory Supplies	EACH
50ml Skirted Centrifuge Tube, Yellow Cap, 25/Bag
TK943-Y	Bio Basic	1PK, 25UNIT
Falcon Conical 15ml Ps Multipack Centrifuge Tubes - PK500
352095	Scientific Laboratory Supplies	PK500
Centrifuge tube 13 ml, flat (800)
TP91019	Westburg	each
15ml Centrifuge Tube, Screw Cap, Red, Sterile, 100/Bag
BT888-SR	Bio Basic	1PK, 100UNIT
15ml Centrifuge Tube, Screw Cap, Blue, Sterile, 100/Bag
BT888-SB	Bio Basic	1PK, 100UNIT
CoolRack XT 5ml holds 12 x 5ml centrifuge tubes - EACH
432060	Scientific Laboratory Supplies	EACH
50 ml Centrifuge Tube with Flat Cap, Bulk, Sterile (20x25)
OLOCT-50B	Westburg	each
15mL Flip Top Centrifuge Tubes Sterile DNase Free - PK500
ARG1154	Scientific Laboratory Supplies	PK500
50mL Flip Top Centrifuge Tubes Sterile DNase Free - PK500
ARG1156	Scientific Laboratory Supplies	PK500
Benchmark Centrifuge Tube Red 5mL (4 bags of 50) - PK200
CEN2048	Scientific Laboratory Supplies	PK200
15ml Centrifuge Tube, Screw Cap, Yellow, Sterile, 100/Bag
BT888-SY	Bio Basic	1PK, 100UNIT
Snap Cap Polyethylene For Centrifuge Tubes Size 2 - PK500
TUB2728	Scientific Laboratory Supplies	PK500
15 ml Centrifuge Tube with Flat Cap, Bulk, Sterile (20x25)
OLOCT-15B	Westburg	each
50 ml Centrifuge Tube with Flat Cap, Racked, Sterile (20x25)
OLOCT-50R	Westburg	each
15ml Centrifuge Tube, Screw Cap, Natural, Sterile, 100/Bag
BT888-SN	Bio Basic	1PK, 100UNIT
Benchmark Centrifuge Tube Blue 5mL (4 bags of 50) - PK200
CEN2046	Scientific Laboratory Supplies	PK200
Benchmark Centrifuge Tube Clear 5mL (4 bags of 50) - PK200
CEN2045	Scientific Laboratory Supplies	PK200
Benchmark Centrifuge Tube Green 5mL (4 bags of 50) - PK200
CEN2047	Scientific Laboratory Supplies	PK200
Benchmark Centrifuge Tube Black 5mL (4 bags of 50) - PK200
CEN2051	Scientific Laboratory Supplies	PK200
15 ml Centrifuge Tube with Flat Cap, Racked, Sterile (20x25)
OLOCT-15R	Westburg	each
50ml High Speed Centrifuge Tube, Natural cap, 25/Bag
CT790-N	Bio Basic	1PK, 25UNIT
Benchmark Centrifuge Tube Yellow 5mL (4 bags of 50) - PK200
CEN2049	Scientific Laboratory Supplies	PK200
50 mL Centrifuge Tube, Rack (PS Rack), Sterile, 25/pk, 500/cs
602051	NEST Biotechnology	500 pcs/cs
50ml Skirted Centrifuge Tube, Green Cap, Sterile, 25/Bag
TK943-GS	Bio Basic	1PK, 25UNIT
Centrifuge tube 16 ml, round, long (800)
TP91017	Westburg	each
50ml Skirted Centrifuge Tube, Yellow Cap, Sterile, 25/Bag
TK943-YS	Bio Basic	1PK, 25UNIT
Centrifuge tube 13 ml, round, short (800)
TP91016	Westburg	each
Corning 15mL PP Centrifuge Tubes Rack Packed with CentriStar Cap Sterile 50/Rack 500/Case - PK500
430790	Scientific Laboratory Supplies	PK500
15ml centrifuge tubes, rack pack, sterile, flat top caps, graduation
SGB011-R	SEEN GROUP LIMITED	50pcs/rack, 500pcs/carton
50ml centrifuge tubes, rack pack, sterile, flat top caps, graduation
SGB012-R	SEEN GROUP LIMITED	25pcs/rack, 500pcs/carton
Kimax Centrifuge tubes graduated beaded top white scale 10ml - PK12
TUB2708	Scientific Laboratory Supplies	PK12
DB 2.1 Single block for 12 x 15ml conical centrifuge tubes - EACH
BLO6032	Scientific Laboratory Supplies	EACH
50 mL Centrifuge Tube, Self-Standing, Bulk, Sterile, 25/pk, 500/cs
602072	NEST Biotechnology	500 pcs/cs
Kimax Reuseable Glass Centrifuge tubes plain high strength 15ml - PK6
TUB2718	Scientific Laboratory Supplies	PK6
50ml High Speed Centrifuge Tube, Natural cap, Sterile, 25/Bag
CT790-SN	Bio Basic	1PK, 25UNIT
MICRO CENTRIFUGE TUBE, AMBER, 5 ML, 250/1250
MCT-500-X	CORNING	250/pk
50ml Conical Centrifuge Tubes, Green Cap, Sterile, 25unit (cardboard rack)
CT788-GS	Bio Basic	1PK, 25UNIT
50ml Conical Centrifuge Tubes, Yellow Cap, Sterile, 25unit (cardboard rack)
CT788-YS	Bio Basic	1PK, 25UNIT
Holder for up to eight 15ml centrifuge tubes, for Bench-Top Shaker
099ASR815	Glascol	each
Holder for up to eight 50ml centrifuge tubes, for Bench-Top Shaker
099ASR850	Glascol	each
Corning 15mL PP Centrifuge Tubes Rack Packed with Plug Seal Cap Sterile 50/Rack 500/Case - PK500
430052	Scientific Laboratory Supplies	PK500
DB 3.1 Single block centrifuge tube combination 4 x 1.5; 3 x 15 ml and 2 x 50ml - EACH
BLO6036	Scientific Laboratory Supplies	EACH
MICRO CENTRIFUGE TUBE, MAXYCLEAR, 5 ML, 250/1250
MCT-500-C	CORNING	250/pk
Centrifuge Tube, Conical, 500mL, PP, sterile, with screw-cap, 8 per bag, 5 bags per case, 40/cs
C1500	MTC Bio	40
Corning 15mL PET Centrifuge Tubes Rack Packed with Plug Seal Cap Sterile 50/Rack 500/Case - PK500
430055	Scientific Laboratory Supplies	PK500
50 mL Centrifuge Tube, Amber Color, Rack (PS Rack), Sterile, 25/pk, 500/cs
602201	NEST Biotechnology	500 pcs/cs
Rack for 4 x 500mL centrifuge tubes, plastic coated wire, autoclavable, 1/ea
C1500-RK	MTC Bio	1
Sterile Centrifuge Tube 5mL (10 bags of 20) - PK200
CEN2053	Scientific Laboratory Supplies	PK200
50ml PP centrifuge tube, free standing, flat screw cap, 25/sterile bag, 500/cs
C2605	MTC Bio	500/pack
30ml PP centrifuge tube, free standing, flat screw cap, 25/sterile bag, 500/cs
C2630	MTC Bio	500/pack
Small centrifuge, 6 x 15 ml, 5000 rpm, incl. Rotor for 6 x 15 ml
CD-0506	Phoenix instrument	each
MICRO CENTRIFUGE TUBE, MAXYCLEAR, 5 ML, STERILE, 250/1250
MCT-500-C-S	CORNING	250/pk
Centrifuge Tube
C1005-T5*-Clear	Benchmark Scientific	1 PC

Next the tube was centrifuged (1000 x g for 5 min.). After centrifugation and separation of  morphotic elements, the obtained plasma was divided into two tubes and frozen at – 85°C until performing laboratory tests.