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--- blood_samples [2023/01/27 13:00] – kevin
+++ blood_samples [2025/02/05 13:49] (current) – external edit 127.0.0.1
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 ====== Blood samples ======
-Fasting blood was collected from all participants aged 8 years or older that participated in [[1A Visit 2]] and [[2A Visit 2]] (and currently in [[3A visit 2]]).\\
+Blood samples are collected in a variety of different blood tubes such as: heparin, EDTA, sodiumfluoride, and gel separation tubes. Blood collected in heparin- and sodiumfluoride tubes are used to perform [[blood_analyses|analyses]] of clinical and haematological parameters (including LDL/HDL cholesterol, HbA1c, and triglycerides) and are therefore no longer available for release. The results of these [[blood_analyses|analyses]] can be obtained upon request.\\
-Blood was collected in various blood tubes (see below), transported from the Lifelines [[location|locations]] to the central lab in Groningen, centrifuged, aliquotted, and stored at -80°C on the day of collection.\\
+Plasma/buffycoat samples derived from EDTA collection tubes and serum samples derived from gel separated tubes are stored in the Lifelines Biobank and can be requested for research purposes.\\
-A subset of plasma samples was used for standard [[blood analyses]]. The remaining samples were placed in long term [[blood samples storage|storage]] to be used by scientists for future analyses.\\
+DNA samples are extracted from the buffycoat fractions and can likewise be requested for research purposes.\\
-Note that [[urine samples]] were collected on the same day.
+\\
+From collection to storage, Lifelines adheres to [[sample quality|stringent quality control]] protocols to ensure that the samples are fit for use in all standard blood analysis protocols. Standard operating procedures for all the steps described below can be provided by Lifelines on request.\\
-From collection to storage, Lifelines adheres to **stringent quality control protocols** to ensure that the samples are fit for use in all standard blood analysis protocols. Detailed SOPs for all the steps described below can be requested from Lifelines if needed.\\
-===== Blood collection =====
+===== Current Study Protocol =====
-All blood samples were collected before 10:00 AM.\\
+Please note that the text below describes the procedures as they are employed during the ongoing [[3A|third]] assessment. The [[1A|baseline]]- and [[2A|second]] assessments differ in various ways, and Lifelines recommends consulting their respective wiki pages to learn more about the procedures employed at those times.
-Blood from fasting adult participants was typically collected using vacuum tubes instead of butterfly needles to avoid the activation of blood enzymes. Blood from fasting children (and in some exceptions from adults) was collected using less invasive butterfly needles.\\
-Blood samples were taken from a blood vessel in the inside of the right elbow, where the veins are relatively close to the surface. In some exceptions the left elbow or the left/right wrist were used.\\
-===== Types of blood tubes =====
+===== Sample Collection =====
-Multiple blood samples were collected in various types of plain glass [[https://en.wikipedia.org/wiki/Vacutainer|Vacutainers®]] to enable all standard blood sample analyses.\\
-  * **Serum samples** are collected in vacutainers without any clot activators, anticoagulants, preservatives or separator material.
+Blood was typically collected between the hours of 07:00 and 10:00, and participants were asked to refrain from consuming any food or liquids (with the exception of water) after 22:00 on the evening preceding visit 2.\\
-  * **Serum gel samples** are collected in vacutainers with a gel composed of inert components, which are part of a polyester-based proprietary formulation. The gel forms a physical barrier between serum or plasma and blood cells during centrifugation.
+Blood was primarily collected through venopuncture of the median cubical vein of the right arm using Venoject® needles in combination with [[https://www.bd.com/en-us/products-and-solutions/products/product-families/bd-vacutainer-blood-collection-tubes#productsaccessories|Vacutainer®]] blood collection tubes, although butterfly needles and venopuncture of other locations (such as the left arm or the wrist) have also been employed if the previous was not an option.
-  * **EDTA samples** are collected in vacutainers with [[https://en.wikipedia.org/wiki/Ethylenediaminetetraacetic_acid|EDTA]] anticoagulants used to prevent clot formation both in vitro and in vivo, in order to maintain blood in the fluid state for hematological testing or to obtain suitable plasma for coagulation and clinical chemistry analyses.
-  * **Heparin samples** are collected in vacutainers with [[https://en.wikipedia.org/wiki/Heparin|sodium heparin]] as an anticoagulant in order to prepare heparinized whole blood.
-  * **Citrate samples** are collected in vacutainers with citric acid, a reversible anticoagulant to enable coagulation assays (only available from [[1A visit 2|1A]]).
-  * **[[https://en.wikipedia.org/wiki/Buffy_coat|Buffy coat]] samples** are collected from the fraction of an anticoagulated blood sample that contains most of the white blood cells and platelets, using density gradient centrifugation.
-{{:20181116_140050.jpg?200|}}
+==== Types of Blood Collection Tubes ====
-===== Centrifuging =====
+The following blood samples are placed into [[blood_samples_storage|cryostorage]] after processing by the Lifelines laboratory.
-| EDTA blood   | 15 min  | 4° C  | 2500RCF  |
+  * Blood (plasma) samples are collected from Lifelines participants (ages 8+) using BD Vacutainer® 10.0 mL K2E (EDTA) 18.0mg Plus blood collection tubes.
-| Serum blood  | 10 min  | 18°C  | 2500RCF  |
+  * Blood (buffycoat) are collected from Lifelines participants (ages 8+) using BD Vacutainer® 10.0 mL K2E (EDTA) 18.0mg Plus blood collection tubes.
-| Citrate      | 10 min  | 4° C  | 2500RCF  |
+  * Blood (serum) samples are collected from Lifelines participants (ages 8+) using BD Vacutainer® 10.0 mL SST II Advance blood collection tubes.
+The following blood samples are used to perform a variety of [[blood_analyses|diagnostic blood tests]].
-{{:dsc_0050.jpg?200|}}
+  * Blood (plasma) samples are collected from Lifelines participants (ages 8+) using BD Vacutainer® 4.5 mL LH (Lithium Heparin) PST™ II Plus blood collection tubes.
+  * Blood (plasma) samples are collected from Lifelines participants (ages 8+) using BD Vacutainer® 4.0 mL K2E (EDTA) 7.2mg Plus blood collection tubes.
-===== Aliquotting =====
+  * Blood (plasma) samples are collected from Lifelines participants (ages 8+) using BD Vacutainer® 4.0 mL NaF 3.0mg Na2EDTA 6.0mg Plus blood collection tubes.
-All blood samples are aliquotted into 900ųl fractions prior to [[Blood samples storage|storage]].\\
+===== Sample Transport =====
-Each aliquot is identified by a unique [[https://en.wikipedia.org/wiki/Data_Matrix| 2D code]].\\
-{{:20181123_135707.jpg?200|}}\\
-Aliquots are followed from creation to storage using the [[https://lifesciences.tecan.com/software-freedom-evoware|Tecan Freedom EVOware® Robot]], a device that optimally combines pipetting, robotics and scheduling of multiple devices in a single package.\\
-{{:tecan.png?200|}}\\
+After collection the blood tubes are placed in a refrigerated (using ice packs), insulated transport box. The temperatures in these boxes are continuously monitored ([[logtag|LogTag TRIX-8]]) as part of our [[sample_quality|quality control]] procedures. These transport boxes are then shipped to the Lifelines laboratory, typically arriving between the hours of 10:00–12:00.
+===== Sample Reception =====
-===== Quality Control =====
+Upon arrival at the Lifelines laboratory the blood collection tubes are divided into two distinct categories:
+  * Blood collection tubes meant for diagnostic testing are refrigerated (at 4 degrees Celsius) and sent onwards to the UMCG in a refrigerated transport box.
+  *  Blood samples to be stored by Lifelines are either immediately centrifuged (according to the protocols detailed below) or refrigerated (at 4 degrees Celsius) until this is possible.
+The temperature measurements taken during transport are reviewed and saved to our labotatory information management system (LIMS).
-Lifelines adheres to stringent protocols and is able to provide for each individual sample detailed information about:
+==== Centrifuging ====
-  * the type of additive in the blood collection tubes
-  * sample processing times and temperatures
-  * hemolysis of the sample
-  * sample storage parameters
-  * the number of freeze-thaw cycles
-During transport, blood samples must remain at a temperature between 0-8°C (cool) or 15-25°C (room temperature). This is checked using a [[LogTag]] temperature reader.
+  * 10.0 mL K2E (EDTA) 18.0mg Plus blood collection samples | 15 minutes @ 2500RCF and 4 degrees Celsius, no brake
+  * 10.0 mL SST II Advance blood collection samples | 10 minutes @ 2500RCF and 18 degrees Celsius, no brake
-{{:20190116_094823.jpg?200|}}\\
+===== Sample Processing =====
-{{:logtag.png?800|}}
+Blood (plasma) and blood (serum) samples are divided into 900 μl fractions by the [[https://lifesciences.tecan.com/freedom-evo-packages|Tecan Freedom EVOware® robot]], a semi-automated system that combines pipetting, robotics, and scheduling of multiple devices.\\
+These aliquots can be identified by a unique 2D barcode printed on the bottom of the tubes, and are held within a 1D barcoded rack ([[https://www.lvl-technologies.com/en/sample-storage/safe-2d-tubes-engl/safe-96-it-2D|LVL technologies Safe® 96 IT]] or equivalent). The exact location of each individual sample is logged and continuously tracked in our LIMS.\\
+During sample processing special care is given to note deviations, such as hemolysis and lipemia, and to register these in our LIMS.\\
+===== Sample Storage =====
+Aliquots are kept refrigerated (at 4 degrees Celsius) until they can be moved into a [[sample_storage|cryofreezer]] (-80 degrees Celsius), which is typically within 10 hours after sample collection.\\