发信人: royluo (roy), 信区: Biology
标 题: 我来贴一个全一点的细胞分离方法的LIST
发信站: The unknown SPACE (Wed May 22 02:01:44 2002) WWW-POST

如果前面那位兄弟只是选克隆的话(比如做stable transfected cell line)，稀释细胞
培养，然后慢慢挑就是。如果要直接从一大堆细胞选出一个符合要求的细胞，方法总结如
下。


From
Culture of animal cells: a manual of basic technique (4th edition)

(1)Sedimentation velocity at 1g
* based on cell size
*Custom-made separating funnel and baffles
*Simple technique but not very high resolution or yield

(2) Isopyknic sedimentation
* based on cell density
*centrifuge
* simple and rapid

(3) MACS
* Surface antibodies
* simple magnet and flow chamber
*specific, given a highly specific surface antibody

(4) Immune Panning
* surface antibodies
* antibody-coated dishes

(5)centrigual elutriation
*cell size, density and surface configuration
*special centrifuge and elutriator rotor
*rapid , high cell yield ,but quite complex process
(6) FACS
*cell surface area fluorescent markers
fluorogenic enzyme substrates, multiparameter
* flow cytometer
* complex technology and expensive, very effective , high
resolution, but low yield

(7)Affinity chromatography
*cell surface antigens, cell surface carbonhydrates
*sterilizable chormatorgraphy column
* elutriation of cells from column is difficult ,better in free
suspension

(8) Countercurrent distribution
*affinity of cell surface constituents for solvent phase
*shaker
* some cells may suffer loss of viability , but method is quite
successful
for others

(9) Electrophoresis in gradient or curtain
* surface charge
* curtain electrophoresis apparatus




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