Hi, I'm Dr. Evan Matthews
I'm here at Montclair State University in the exercise science lab to show you how to take a blood sample, and either turn it into
a serum or plasma sample that can be either used or stored. So anytime you're handling a blood sample or other similar
biohazard make sure that you're wearing the appropriate clothes. So when you are working with a biohazard like blood
you need to make sure that you're protecting yourself from that biohazard.
So you need to be wearing shoes that are closed.
So that there is no exposure of your feet. You need to be wearing pants that
prevent an exposure of your legs. You should be wearing some sort of lab coat that protects most of the rest of your body.
Then you need to be wearing gloves that protect you from anything
that's going to get spilled on your hands, and you need to be wearing some sort of protective eyewear.
That's going to have some side shields as well as a lot of coverage to your eyes to prevent any
splashes from getting to your eyes. So the type of blood samples we're talking about today are going to be collected either through a
typical venipuncture, just using a needle or an IV catheter.
Which is also type of venipuncture either way we're going to be collecting
tubes that are typically going to be either a peripheral top or red top the purple top are the
EDTA tubes that are going to prevent the blood from coagulating
The red tops are gonna be the silicone coda tubes that are going to promote coagulation of that blood
So whether you prevent coagulation or promote coagulation is going to determine? Whether you get a?
Plasma, or serum all right plasma is what you get from the purple top tubes
Where you prevent the coagulation?
So it still has platelets in it serum is what you're going to get from the red top tubes
Where you promote a coagulation?
So essentially that coagulation is the clotting of those platelets?
And so when you spin this down you're going to end up getting just liquid and no
Platelets inside that liquid so either way you're going to take the blood tube
When you first get it and you're going to invert it very gently and very slowly typically between four and six times
and that's going to make sure that the blood has been thoroughly mixed with whatever type of treatment is inside that tube again either a
coagulant or an anticoagulant
And then you're going to take the blood and you're going to set it up right in a tube holder and allow the blood to
If it is a coagulating tube, or you can spin it right away if it's a non-coagulating tube. Alright so one mistake
I made that you're gonna see in this video is that
I forgot so you put them in a tube holder
It doesn't really make a big difference
But it makes it hard to tell whether or not the blood has been coagulated because how you sell blood has been
Coagulated if you take it you very gently turn it upside down
And if the blood doesn't move if it doesn't flow downward
Then that means it's coagulated, and you can still see that with it being on its side
It's just not quite as clear alright
So once you have confirmed that your coagulated blood or the blood that you want to coagulate has coagulated
Then you can take the blood samples, and you can put it in a centrifuge alright centrifuges can be
Set to several different parameters
You can set the temperature of the centrifuge you can make it room temperature
Which is approximately?
what I'm going to be setting it at or you can make it cold which often times needs to be done if you're running a
sample for hormones or something that's going to
Degrade quickly at room temperature the other things you can set are of how fast it's going to spin
I'm going to spin
It just beyond 3000 RPM's
Or you can set also the time of the spin, which is something that typically is between 10-15 minutes
And you can decide whether or not to have a brake applied
Or how heavily to apply that brake at the end of the centrifuge time period.
Set those parameters to whatever type of analysis you're going to be doing and
Then you're going to put the blood tubes inside the centrifuge now make sure that you're putting it inside
The proper tubes within the centrifuge so if you put it in two or two holders that are too large for the tubes
It's not going to hold it very well
And you don't want that so put it in the right size tube holders for the tube you're using
All right
So the other thing you need to do when you're putting the tubes inside the centrifuge is try to
Disperse them in such a way
Or so you place them in there in such a way that they are balancing each other and their weights
So if you have one over here have one over there alright, so that when it's spinning
It's going to have the same weight on both sides of the inside of that Center huge
And it's gonna promote an even spinning so once the tubes are in the centrifuge appropriately
Press the lid down until it clicks move over the locking mechanism on the lid
and then you're simply gonna turn the centrifuge all if you haven't already and
Hit the button if there is a button to start the spin
so this particular
centrifuge does not have a button to spin you just turn the
The timer to whatever you want to be set at and that's going to initiate the spinning process so you're gonna see a very slow
In increase in the RPM's of the device because
It doesn't want to start super super quickly it's gonna sort of ramp up and that's gonna prevent it so much
jostling of the blood inside this the tubes which is something you don't want you don't want the
Tubes being sort of jostle around because it's gonna increase the likelihood of damaging the sample, okay?
so once the centrifuge is done its going to
Start to break itself if you have the brake applied you can see that the RPM's that are going to slow down
Fairly quickly if you don't have the brake apply it takes a very long time for the centrifuge to stop
All right, so once the centrifuge has stopped
You can open it, so you're simply going to flip the switch back on the locking mechanism on the lid
And then there's a button on the front of the centrifuge that you have to push to actually open the lid
So there's two different locks
So once both those locks are undone you can lift the lid take the samples out and you can use them
However you plan on using them
so when you're taking the
Tubes out of the centrifuge and to wherever your workstation is or when you're working with them at the workstation
Make sure that you never invert those tubes again
So if you invert the tubes you're going to see that the blood remix is partially and essentially you're ruining your sample by doing that
So if you look at
tubes actually they've been centrifuge there's going to be a clear separation between the blood cells at the bottom and
The clear liquid on top so what that clear liquid is it depends on the type of tube whether it is a coagulant or an?
anticoagulant tube so again the
anticoagulant tubes are going to have plasma in them so that liquids gonna still contain platelets the
coagulating tubes are gonna have serum in them because all the platelets have coagulated and been spun to the
Closer to the cells in the bottom the red blood cells at the bottom alright, so either way you're going to take those tubes
You're going to very gently remove the top to just kind of wiggle it back and forth until it's able to
pop off and
Then you're going to take is some sort of pipette, so we're just going to be using a simple bulb transfer pipette today
And you're going to suck up that
Serum or plasma and transfer it into
Some sort of container to be used in a machine or to be stored in a freezer all right, so
When you're sucking up that?
Serum or plasma though start the very top so start at the top the plasma and slowly work your way down
As the plasma is sucked up into the bulb pipette so the reason you're doing that is if you get too close to the cells
About midway down the tube you're going to disturb those cells and in doing so it's going to remix
The tube of blood and it's going to essentially destroy your sample so start at the top very conservatively
stuck a little bit of the of the serum or plasma and continue to lower the pipette as you're sucking up serum or plasma and
Stop when either you fill the pipette, or if you get too close to the cells where you start thinking that maybe you might?
Suck a little bit of the cells up and mix your sample so stop before that happens then
then take that sample put it wherever you're going to use a
Store or use it
And then if you need more you can go back and try to get a little bit more serum or plasma
But you want to make sure that you get the easy serum and plasma first before you start getting a little more gutsy and getting
The serum or plasma that's closer to the cells
So once you have
Transferred all of your serum or plasma to another container that you're going to use for analysis or for storage
You will have officially aliquoted the serum plasma
So if you're going to store it make sure that goes into a refrigerator or freezer more likely and that
Happens quickly because anytime that the this that the sample is at room temperature
It is degrading the sample so make sure that you get it into its storage location as quickly as you can
Alright so that was a really quick overview on how to
Spin blood and pull off out of the serum or plasma for storage or use you have any questions
Please put those in the comments below, and I'll try to get back to you otherwise please come back and watch another video. Thanks