Method and means for sorting individual particles into containers for culturing, cloning, analysis, or the like

In a method of separating individual particles from other particles contained in a fluid stream and maintaining the separation without pooling of certain particles the combination comprising,

• producing a flow stream which includes particle-containing fluid,
• jetting the fluid flow stream,
• modulating the jet stream velocity in the direction of the stream axis to break the stream into discrete drops,
• detecting particles in the stream and producing a sort signal in response to the detection of certain particles,
• triggering operation of a pulse generator by a time delayed sort signal for charging the jet stream while drops which contain a detected certain particle break from the jet stream,
• deflecting charged drops in an amount related to the drop charge,
• in response to production of a sort signal, temporarily preventing production of subsequent sort signals, and
• continuing for an indefinite period of time to prevent production of subsequent sort signals whenever a delayed sort signal triggers operation of the pulse generator.

Particle sorting apparatus for loading containers with single particles for culturing, cloning, or the like, comprising,

• a nozzle,
• means for jetting a stream of particle-containing fluid from the nozzle,
• means for modulating the velocity of the stream to break the jet stream into discrete drops,
• detector means for detecting particles in the stream,
• pulse charging means for supplying drops with different drop charges,
• means for deflecting charged drops an amount related to the drop charge,
• a stream catcher for catching drops not separated from the drop stream,
• manually controlled means for enabling operation of the pulse charging means to render the same responsive to the detector means for sorting a single detected particle contained in a drop from the stream of drops,
• means for automatically disabling operation of the pulse charging means following sorting of the single particle from the stream,
• a movable tray support for the support of a multiwell tray into which drops separated from the drop stream are sorted, and
• releasable detent means for releasably locking said tray support at a plurality of spaced positions for catching drops separated from the drop stream in the tray wells.

A tray supporting mechanism for movable support of a multiwell tray for use with a particle sorting system of the type which includes means for producing a stream of discrete drops, some of which drops contain particles to be sorted from the stream by separation of certain particle-containing drops from the drop stream, said mechanism comprising

• a base toward which said drop stream is directed,
• carriage means on the base movable in at least one direction therealong,
• a tray holder attached to said carriage means for support of a multiwell tray at a fixed location on the holder, and
• releasable detent means for releasably locking said carriage means at spaced positions along said at least one direction for positioning tray wells to receive drops separated from the drop stream,
• said carriage means being manually movable for manual positioning thereof and the attached tray holder.

A tray supporting mechanism for movable support of a multiwell tray for use with a particle sorting system of the type which includes means for producing a stream of discrete drops, some of which drops contain particles to be sorted from the stream by separation of certain particle-containing drops from the drop stream, said mechanism comprising

• a base toward which said drop stream is directed,
• carriage means on the base movable in at least one direction therealong,
• a tray holder attached to said carriage means for support of a multiwell tray at a fixed location on the holder, and
• releasable detent means for releasably locking said carriage means at spaced positions along said at least one direction for positioning tray wells to receive drops separated from the drop stream,
• said tray holder being attached to said carriage means by releasable fastening means for adjustable positioning of the tray holder along at least said one direction relative to said carriage means in the released condition of said fastening means for initial alignment purposes.

A tray supporting mechanism for movable support of a multiwell tray for use with a particle sorting system of the type which includes means for producing a stream of discrete drops, some of which drops contain particles to be sorted from the stream by separation of certain particle-containing drops from the drop stream, said mechanism comprising

• a base toward which said drop stream is directed,
• carriage means on the base comprising a Y carriage movable along the base and an X carriage movable along the Y carriage in perpendicular Y and X directions, respectively,
• a tray holder attached to said carriage means for support of a multiwell tray at a fixed location on the holder, and
• releasable detent means for releasably locking said carriage means at spaced positions along said X and Y directions for positioning tray wells to receive drops separated from the drop stream, said releasable detent means including a first releasable detent for releasably locking said Y carriage at spaced positions along the Y direction and a second releasable detent for releasably locking said X carriage at spaced positions along the X direction, said first releasable detent including means for releasably locking said Y carriage at positions numbering one-half the total number of rows of tray wells and said second releasable detent means includes means for releasably locking said X carriage at positions numbering the total number of columns of tray wells.

A tray supporting mechanism for movable support of a multiwell tray for use with a particle sorting system of the type which includes means for producing a stream of discrete drops, some of which drops contain particles to be sorted from the stream by separation of certain particle-containing drops from the drop stream, said mechanism comprising

• a base toward which said drop stream is directed,
• carriage means on the base comprising a Y carriage movable along the base and an X carriage movable along the Y carriage in perpendicular Y and X directions, respectively,
• a tray holder attached to said carriage means for support of a multiwell tray at a fixed location on the holder, and
• releasable detent means for releasably locking said carriage means at spaced positions along said X and Y directions for positioning tray wells to receive drops separated from the drop stream, said releasable detent means including a first releasable detent for releasably locking said Y carriage at spaced positions along the Y direction and a second releasable detent for releasably locking said X carriage at spaced positions along the X direction, said second releasable detent means including means for releasably locking said X carriage at an additional position for initial alignment of the tray holder relative to drops separated from the stream.