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    Lumbar Puncture

    Clemens Scherzer, Bradley Hyman, Charles Jennings

    Published: 2022-11-22 DOI: 10.17504/protocols.io.q26g78x11lwz/v1
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    Abstract

    This protocol explains the Standard Operating Protocol for performing a Lumbar Puncture.

    Before start

    *Optimum time delay between withdrawal and freezing should be within 1-2 hours per European consensus.

    Ref:

    Citation
    Teunissen CE, Petzold A, Bennett JL, Berven FS, Brundin L, Comabella M, Franciotta D, Frederiksen JL, Fleming JO, Furlan R, Hintzen RQ, Hughes SG, Johnson MH, Krasulova E, Kuhle J, Magnone MC, Rajda C, Rejdak K, Schmidt HK, van Pesch V, Waubant E, Wolf C, Giovannoni G, Hemmer B, Tumani H, Deisenhammer F 2009 A consensus protocol for the standardization of cerebrospinal fluid collection and biobanking. Neurology http://doi.org/10.1212/WNL.0b013e3181c47cc2

    Steps

    Lumbar Puncture

    1.

    Label the collection tubes with the sample ID as appropriate.

    2.

    Place aliquot tubes on dry ice prior to procedure so they are pre-cooled.

    3.

    Perform lumbar puncture using the atraumatic technique, inserting the needle with the bevel in parallel to the dura fibers. If bloody, discard the first 0.5 mL to clear any blood then collect. Record time of lumbar puncture.

    4.

    Collect 1mL into one 14 mL polypropylene tube at Room temperature for routine diagnostic labs.

    5.

    Collect 20mL into two 10 mL syringes at Room temperature. Then, transfer all 20 ml into one 50 ml Falcon tube, mix gently by inverting 3-4 times.

    6.

    Place 0.5mL in one transfer tube for routine cell count, and 0.5mL in another transfer tube for glucose and total protein. Send at Room temperature to core lab with completed requisition form.

    Note
    Send to local clinical lab for basic CSF analyses - must be analyzed within 4 hours of collection . Cell count (500 µl) Total protein and Glucose (500 µl)

    7.

    Spin at 400x g right after CSF is collected. Record time of spinning.

    8.

    Pipet 0.5mL into the remainder (34-36 aliquots) 1.5 mL pre-cooled Fisherbrand silconized polypropylene transfer tubes.

    9.

    Freeze aliquots immediately on dry ice and transfer to Scherzer lab.

    10.

    Identify samples with obvious blood contamination.

    11.

    Scan specimens and place in -80°C freezer within 2 hours. Record time of freezing.

    12.

    Time of LP collection, spinning and freezer time will be recorded and entered into freezerworks.

    13.

    Specimens will be scanned, frozen, and stored in two separate freezers at the Scherzer lab.

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