What is a Ficoll gradient?

What is a Ficoll gradient?

Ficoll 400 is completely non-ionic. The most common application for Ficoll 400 is as a density gradient medium for the separation and isolation of eukaryotic cells, organelles and bacterial cells. Density ranges up to 1.2 g/ml can be attained. It has also been utilized in a variety of other applications.

How do you make a Ficoll gradient?

This is my method:

1. Gently homogenize the blood sample inside heparin blood collection tube.
2. Add the whole blood to conical tube that contain 4 ml of PBS (equal volume to the sample; 1:1)
3. Homogenize or mix the solution.
4. Add the entire solution to a new conical tube containing 4ml ficoll solution.

What is Ficoll density gradient centrifugation?

Ficoll density gradient centrifugation is one of the most commonly used method for isolation and enrichment of human mononuclear cells, particularly from peripheral blood and other biological fluids, e.g., umbilical cord blood and bone marrow [1], [2].

What is density of Ficoll-Hypaque solution?

Ficoll-Paque PREMIUM (density 1.077 g/ml) is a density gradient media used as a reagent in clinical research and cell therapy manufacturing applications for in vitro isolation of human mononuclear cells. These may be used when higher or lower densities than the standard 1.077 g/ml are required.

What is a density gradient used for?

Density gradient is a spatial variation in density over an area. The term is used in the natural sciences to describe varying density of matter, but can apply to any quantity whose density can be measured.

How does density gradient centrifugation work?

Isopycnic gradient centrifugation occurs when centrifugation continues until all the particles in the gradient have reached a position where their density is equal to that of the medium. This type of centrifugation separates different particles based on their different densities.

Why is Ficoll used?

Ficoll is a neutral, highly branched, high-mass, hydrophilic polysaccharide which dissolves readily in aqueous solutions. Ficoll is part of Ficoll-Paque, which is used in biology laboratories to separate blood to its components (erythrocytes, leukocytes etc.).

How do you use Ficoll Paque?

1. Invert the Ficoll-Paque media bottle several times to ensure thorough mixing.
2. Add Ficoll-Paque media (3 mL) to the centrifuge tube.
3. Carefully layer the diluted blood sample (4 mL) onto the Ficoll-Paque media solution (Figure 3).
4. Centrifuge at 400 g for 30 to 40 min at 18 ºC to 20 °C (brake should be turned off).