In this Video, we aim to show a technique of blood collection in zebrafish. This technique needs a container for anesthesia, a Petri dish, and a scalpel, low rotation tips and automatic micropipet, P 20 and micro tubes, and the centrifuge. Before I Start the blood collection, it's necessary to anesthetize the fish with a fishing net.
The fish is moved from the aquarium to a container that has a aquarium water and ice chips. The time of Anesthesia depends on each fish. It usually takes from three to six Seconds.
The fish is considered anesthetized. When it does not respond to a sternal stimulus, the fish has to be gently transferred from anesthesia container to a dry gauze to remove the excess of water. When the fish Is anesthetized and the excess of water is removed, the fish is gently involved in a dry gauze that covers body and head, leaving free access to the tail.
With a scalpel, a transverse incision is done just before the tail and the blood starts to Flow. The Collected Blood is then replaced into a micro tube. The tube should be gently handled to avoid hemolysis.
Once the blood is collected, it's necessary to centrifuge to obtain the serum. It is possible to use a pool of blood samples since the time between the first blood collection and the centrifuge do not exceed 10 minutes. Moreover, the blood should be centrifuged for 10 minutes.
These steps should be rigorously followed in order to avoid demo moly of the samples I.