Multiple Sclerosis International / 2011 / Article / Tab 2

Review Article

Consensus Guidelines for CSF and Blood Biobanking for CNS Biomarker Studies

Table 2

Guidelines for procedure of CSF withdrawal.

Item no.ProcedureIdeal situation

(A) Collection procedures
(1)Preferred volumeAt least 12 mL. First 1-2 mL for basic CSF assessment (item 26). Last 10 mL for biobanking. Record volume taken and fraction used for biobanking, if applicable.
(2)LocationVertebral body L3–L5
(3)If bloodyDo not process further.
Criteria for bloody: more than 500 red blood cells/μL.
Record number of blood cells in diagnostic samples.
(4)Type of needleAtraumatic
(5)Type of collection tubePolypropylene tubes, screw cap, volume >10 mL.
(6)Time of day of withdrawal and storagePreferably standardized within each center allowing for intercenter differences in local logistics.
Record date and time of collection.
(7)Other body fluids that should be collected simultaneouslySerum
(8)Other body fluids that should be collected simultaneouslyPlasma: EDTA (preferred over citrate).

(B) Processing for storage
(9)Storage temperature until freezingRoom temperature before, during, and after spinning.
(10)Spinning conditionsSerum: 2,000 g, 10 min at room temperature.
CSF: 400 g, 10 min at room temperature/2,000 g if no cells are to be preserved.
(11)Time delay between withdrawal and spinning and freezingOptimal for CSF: 1-2 hours
Optimal for serum: 30–60 min.
Thus doing “both body fluids simultaneously”: ideally within one hour.
After spinning, samples must be aliquoted and frozen immediately for storage at −80ºC.
(12)Type of tube for aliquotingSmall polypropylene tubes (1 to 2 mL) with screw caps. Record manufacturer.
(13)AliquotingA minimum of two aliquots is recommended. The advised research sample volume of 10 mL should be enough for >10 aliquots.
(14)Volume of aliquotsMinimum 0.1 mL. Depending on total volume of tube: 0.2, 0.5, and 1 mL. Preferably, the tubes are filled up to 75%.
(15)CodingUnique codes. Freezing-proof labels. Ideally barcodes to facilitate searching, to aid in blinding the analysis and to protect the privacy of patients.

(C) Storage conditions and administration
(16)Freezing temperature−80ºC
(17)Additional items on sample collection protocols that must be recordedLocation of samples
(18)Additional items on sample collection protocols that must be recordedSurveillance of freezers
(19)Additional items on sample collection protocols that must be recordedSplitting of samples over two or more freezers
(20)Transport conditionsAlways on dry-ice, sufficient volume of dry-ice for minimal 3 days of transport.
Initiated on Mondays.
Avoid high temperatures for thawing and mix thoroughly.

Article of the Year Award: Outstanding research contributions of 2020, as selected by our Chief Editors. Read the winning articles.