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Dear all,
I am very much thankful if any one tell me how to collect 1min.,
2min., 3min., 4min., and 5 min., blood samples from rat without
necessity of cannulation.
thank you
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The following message was posted to: PharmPK
Dear Krishna,
You can use anesthetics agents i.e. pentobarbitone, where you can get
the animal unconscious for at least 40-50 min but first you should
check the interference of the anesthetics agents to drug. The route
you can use is either retro orbital or sub lingual vein.
Regards,
Chintan
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The following message was posted to: PharmPK
Hi,
You can do this using saphaneous vein puncture or
alternatively by tail vein bleed (provided you are not
administering drug through tail). For saphaneous you
need to restrain rat in a jig, take out and hold the
leg between your fingers, shave of the hair using
suitable hair clipper, identify the saphaneous vein,
apply silicone grease and puncture using 21 or 23G
needle. This technique will not give you lot of blood
like cannulation or retro orbital method. We routinely
use this technique to do PK. Our sample volume at each
time is 25ul (6-8 time points by alternating legs)
collected using heparinised glass capillary, out which
we easily get 10ul of good quality plasma. Our
analytical methods (QQQ) just require 10ul of plasma.
This method is fast, convenient, does not require
anaesthetics and surgical manipulation however do
require some practice initally.
Hope this helps!
Jagdish
--
Dr Jagdish Kumar Jaiswal
Research Fellow
Auckland Cancer Society Research Centre
FMHS, The University of Auckland
Private Bag 92019
Auckland 1142, New Zealand
Phone +64 9 3737599 ext 86090
Fax +64 9 3737571
email j.jaiswal.-at-.auckland.ac.nz
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Tail vein or orbital bleeding. Tail vein, leave needle in place and
after sample use heparinizes saline to prevent clotting. Orbital
bleeding can get either the 2 and 4 min sample from one rat and 3 and
5 min from another.
Stanley Cotler
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The following message was posted to: PharmPK
Greetings,
Yes, back to back early blood collections can impose a bit of a
technical challenge.
Perhaps you can use a cohort study design (group A = 1 and 4 min, group
B = 2 and 5 min, etc...)
-or-
Develop the skill of bleeding rats from the jugular vein using a manual
restraint. With practice (and patience!) you will be able to draw blood
collections less than one minute apart (even between different animals).
The procedure requires no anesthesia, no restraining devices and I feel,
because of it's quickness, puts less stress on the animals. Once a
technician has accomplished the skill, the procedure is as simple as
picking up the rat in one hand (in a very specific hold) and collecting
blood with the other (recommended: 1cc syringe w/ 3/4" 23g needle).
Also, the procedure won't interfere with IV tail dosing, and doesn't use
any anticoagulant coated capillary tubes (in case that might interfere
with your assay).
Typical draw volumes range from 0.25mL to 1mL with this technique.
Good luck!
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Hi,
Maybe you just need to expose the blood vessel you wanted and clamp it
with a hemostat, and then collect the blood samples at your designated
time points.
Hope this be useful!
One more thing\0xFF0CPlease take care of the welfare of animals!
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