Method and apparatus for manipulating single cells and small aggregates thereof

Abstract

A method for using a well, in particular a well open at an upper end and having a vertical axis, able to contain a liquid and particles contained in said liquid, includes the use of said well in a normal configuration with said upper end facing upwards, and in an overturned configuration with said upper end facing downwards in which a liquid is retained in the well by capillarity.

1 . A method for using a well, in particular a well open at an upper end and having a vertical axis, able to contain a liquid and particles contained in said liquid, comprising the use of said well in a normal configuration with said upper end facing upwards, and in an overturned configuration with said upper end facing downwards in which a liquid is retained in the well by capillarity. 
     
     
         2 . The method of  claim 1 , wherein the well comprises at least two manipulation electrodes able to be powered by electrical voltages, the method further comprising applying an alternating voltage to at least one of said electrodes to induce dielectrophoresis effects on at least one particle contained inside said well. 
     
     
         3 . The method of  claim 2 , wherein the at least two manipulation electrodes comprises at least three electrodes disposed in parallel planes and forming an annular rim, the method further comprising applying a substantially in-phase sinusoidal voltage to two electrodes, and a substantially counterphase voltage to a third electrode, or voltages suitable for forming an analogous electric field inside the well. 
     
     
         4 . The method of  claim 2 , wherein at least one of said electrodes forms an annular rim along the delimiting wall of the well, the rim being disposed substantially in a plane transverse to the axis of symmetry of the well, wherein the well comprises at least a pair of electrodes disposed in the same plane transverse to the vertical axis of symmetry of the well, and forming two annular rim portions along the delimiting wall of the well, separated from each other by a surface parallel to the axis of the well wherein said pair of electrodes and said electrode forming an annular rim are disposed in different transverse planes, with said electrode forming an annular rim disposed towards the upper end of the well with respect to said pair, the method further comprising applying a counterphase sinusoidal voltage to the electrodes of said pair, and applying a zero voltage to the remaining electrode, or voltages able to form an analogous electric field inside the well. 
     
     
         5 . The method of  claim 2 , wherein at least one of said electrodes forms an annular rim along the delimiting wall of the well, the rim being disposed substantially in a plane transverse to the axis of symmetry of the well, the well comprising two pairs of electrodes disposed in two difference planes transverse to the axis of the well, the method further comprising applying a substantially counterphase sinusoidal voltage to the electrodes of each of said pairs so that the voltages of the two electrodes superimposed in the direction of the well axis are also substantially in counterphase, or voltages able to form an analogous electric field inside the well. 
     
     
         6 . The method of  claim 2 , further comprising a final step of isolated cell separation. 
     
     
         7 . The method of  claim 2 , further comprising measuring, using a measuring sensor, an impedentiometric, optical or potentiometric signal associated with the at least one particle or with the culture medium in the well or a variation thereof. 
     
     
         8 . The method of  claim 7 , where said optical signal is a fluorescence, luminescence or optical density signal. 
     
     
         9 . The method of  claim 7 , wherein said signal or said variation identifies the content or concentration of lipid in the at least one particle or culture medium. 
     
     
         10 . The method of  claim 7 , where said signal or said variation identifies an isolated cell lysis. 
     
     
         11 . The method of  claim 2 , wherein the particles are subjected to an electric field suitable for electroporation. 
     
     
         12 . The method of  claim 2 , comprising forcing particles to interact. 
     
     
         13 . The method of  claim 12 , further comprising eliciting electrofusion of the particles. 
     
     
         14 . The method of  claim 7 , comprising confining a particle by dielectrophoresis and changing the liquid in the well. 
     
     
         15 . The method of  claim 7 , wherein the at least one particle is an isolated cell, a lipid vesicle or a liposome. 
     
     
         16 . The method of  claim 15 , wherein said isolated cell is a eukaryotic cell, a microorganism or a protoplast. 
     
     
         17 . The method of  claim 16 , wherein said microorganism is a yeast. 
     
     
         18 . The method of  claim 16 , wherein said microorganism is a bacterium. 
     
     
         19 . The method of  claim 1 , wherein the well is disposed in a platform comprising at least one additional well. 
     
     
         20 . The method of  claim 2 , wherein the alternating voltage is sinusoidal.