cardiac puncture

2013-12-01

azim58 - cardiac puncture

Cardiac Puncture

also located here
"S:\Research\Cancer_Eradication\Discovering tumor specific
antigens\Protocols\Cardiac Puncture.docx"

http://www.youtube.com/watch?v=cRx0E-bu95Q
http://waweisslab.ucsf.edu/display_protocol.php?PTCID=o2uir1ozi1

Protocol from Current Protocols in Immunology
http://onlinelibrary.wiley.com/doi/10.1002/0471142735.im0107s73/pdf

see also a protocol from Deborah Hansen
"F:\kurt\storage\CIM Research Folder\DR\2013\5-8-13\spleen and blood
collection protocols\20130507 final bleeds + spleen.xlsx"

CARDIAC PUNCTURE OF MOUSE AND RAT
Materials

1- to 3-ml syringe (mouse) or 3- to 10-ml syringe (rat), with 20- to

22-G needle

Serum separator BD Microtainer tube for collecting blood

Additional reagents and equipment for anesthesia (UNIT 1.4)

Anesthetize the mouse or rat (most likely with avertin). Place the

animal in dorsal recumbency.

Before inserting the needle, pull the plunger back and forth to

loosen everything up.

Insert needle just below and slightly to the left of the xiphoid

cartilage at the base of the sternum, at a 15◦ to 20◦ angle (mouse)
or 20◦ to 30◦ angle (rat) (Fig. 1.7.4). In my experience, the
needle often does not need to be inserted very far.

Advance the needle slowly, applying very slight negative pressure on

the barrel of the syringe (blood will flow into the hub of the needle
when the tip has entered one of the chambers of the heart). Aspirate
gently until blood flow ceases. Advancing or retracting the needle tip
may be necessary to obtain a maximal volume. Also slightly rotating the
needle slightly seems to help greatly in some situations.

Remove the needle from the syringe and transfer the blood from the

syringe to a BD microtainer serum separator tube (Blood Separation
Tubes). Note that the blood should not be passed through a needle
because this can damage some components in the blood.

Note that once obtaining blood from the heart becomes too difficult,

a fair amount of blood from the punctured heart can be collected from
the bottom of the body cavity.

Ensure death by cervical dislocation (UNIT 1.4) when collection is

complete.

If it is absolutely necessary, more blood can be collected by cutting

the mouse open and collecting blood from the bottom of the cavity (it
may help to snip the heart)

Note:
I think Andrey used a 25 guage needle one time
(smaller numbers indicate larger holes)