Microlithographic array for macromolecule and cell fractionation

A sorting apparatus for fractionating microstructures, in a fluid medium, said sorting apparatus comprising:

• a. a substrate having a receptacle located on a side thereof, said receptacle having first and second ends and a floor bound on opposite sides by a pair of upstanding opposed side walls extending between said first and second ends of said receptacle, migration of the microstructures from the first end of said receptacle to said second end of said receptacle defining a migration direction for said receptacle;
• b. sifting means positioned within said receptacle intermediate said first and second ends thereof traversing said migration direction for interacting with the microstructures to partially hinder migration of the microstructures in said migration direction in the fluid medium, said sifting means comprising an array of obstacles upstanding from said floor of said receptacle, and arranged in a preselected and reproducible pattern, each of said obstacles having:
• (i) a v-shaped cross section in a plane disposed parallel to said floor of said receptacle, said v-shaped cross section of said obstacles comprising first and second arms intersecting at an end of each to form a vertex of said v-shaped cross section, the other ends of said first and second arms remote from said vertex forming an open end of said v-shaped cross section;
• (ii) a top at the end thereof opposite from said floor of said receptacle;
• (iii) a height measured in a direction normal to said floor of said receptacle;
• (iv) a length measured in a direction parallel to said migration direction;
• and
• (v) a width measured in a direction normal to said migration direction,
• (vi) the distance of each of said obstacles from an adjacent of said obstacles defining a separation distance therebetween;
• and
• c. ceiling means positioned over said sifting means for covering said receptacle and for causing migration of the microstructures in said receptacle to occur through said sifting means exclusively.

A sorting apparatus for fractionating microstructures in a fluid medium, said sorting apparatus comprising:

• a. a substrate having a receptacle located on a side thereof, said receptacle having first and second ends and a floor bound on opposite sides by a pair of upstanding opposed side walls extending between said first and second ends of said receptacle, migration of the microstructures from the first end of said receptacle to said second end of said receptacle defining a migration direction for said receptacle;
• b. sifting means positioned within said receptacle intermediate said first and second ends thereof traversing said migration direction for interacting with the microstructures to partially hinder migration of the microstructures in said migration direction in the fluid medium, said sifting means comprising an array of obstacles upstanding from said floor of said receptacle and arranged in a preselected and reproducible pattern, each of said obstacles having:
• (i) a cup-shaped cross section in a plane disposed parallel to said floor of said receptacle, said cup-shaped cross section of said obstacles comprising:
• (A) a first leg and a second leg positioned in opposed relationship substantially parallel to said migration direction, each of said first and second legs having thereby oppositely disposed first and second ends;
• and
• (B) a third leg positioned substantially perpendicular to said migration direction and being connected between said first ends of each of said first and second legs, thereby to define between said second ends of said first and second legs an open end of said cup-shaped cross section;
• (ii) a top at the end thereof opposite from said floor of said receptacle;
• (iii) a height measured in a direction normal to said floor of said receptacle;
• (iv) a length measured in a direction parallel to said migration direction;
• and
• (v) a width measured in a direction normal to said migration direction,
• (vi) the distance of each of said obstacles from an adjacent of said obstacles defining a separation distance therebetween;
• and
• c. ceiling means positioned over said sifting means for covering said receptacle and for causing migration of the microstructures in said receptacle to occur through said sifting means exclusively.

An apparatus for sorting microstructures in a fluid medium, said apparatus comprising:

• a. a substrate having a receptacle located on a side thereof, said receptacle having first and second ends and a floor bound on opposite sides by a pair of upstanding opposed side walls extending between said first and second ends of said receptacle, the height of said side walls defining a depth of said receptacle;
• b. a two-dimensional array of obstacles upstanding from said floor of said receptacle, each of said obstacles in said array having a height measured in a direction normal to said floor of said receptacle, said height of said obstacles being substantially equal to said depth of said receptacle;
• and
• c. ceiling means positioned over said array of obstacles for covering said receptacle and for causing the migration of the microstructures within said receptacle to occur exclusively through said array of obstacles, the end of each of said obstacles remote from said floor of said receptacle engaging said ceiling means, thereby to preclude migration of the microstructures between said ceiling means and each of said obstacles.

A reusable apparatus for sorting individual microstructures in a fluid medium, said apparatus comprising:

• a. a substrate having a receptacle located on a side thereof, said receptacle having first and second ends and a floor bound on opposite sides by a pair of upstanding opposed side walls extending between said first and second ends of said receptacle, the height of said side walls defining a depth of said receptacle, said depth of said receptacle being commensurate with the size of the microstructures in the fluid medium, whereby when the microstructures are caused to migrate in the fluid medium from said first end through said second end of said receptacle the microstructures do so in essentially a single layer;
• and
• b. a two-dimensional array of obstacles positioned in said receptacle intermediate said first and second ends thereof, said obstacles being comprised of a rigid self-sustaining material that is chemically inert to the microstructures and to the fluid medium in a normal range of temperatures suitable for fractionating the microstructures;
• and
• c. capping means positioned over and contacting said array of obstacles for affording visual observation of migration of the microstructures in said migration direction through said receptacle, and for precluding migration of the microstructures between said array of obstacles and said capping means.

An apparatus for channelling cells through passageways and simultaneously viewing the cells in a fluid medium, said apparatus comprising:

• a. a substrate having a receptacle located on a side thereof, said receptacle having a floor bound by a pair of upstanding opposing side walls and a first end and a second end, the height of said side walls defining a depth of said receptacle, said depth of said receptacle being commensurate with the size of the cells in the fluid medium, whereby when the cells are caused to migrate in the fluid medium through said receptacle, the cells do so in essentially a single layer, the migration of cells from the first end to the second end of said receptacle defining a migration direction for said receptacle;
• b. a two-dimensional array of obstacles positioned in said receptacle intermediate said first and second ends thereof, said array of obstacles allowing passage of the cells through said receptacle in at least one single file;
• and
• c. capping means for covering said receptacle intermediate said first and second ends thereof and for affording visual observation of the migration of the cells through said array of obstacles and through said receptacle.

An apparatus for channelling cells through passageways and simultaneously viewing the cells in a fluid medium, said apparatus comprising:

• a. a substrate having a receptacle located on a side thereof, said receptacle having a floor bound by a pair of upstanding opposing side walls and a first end and a second end, the height of said side walls defining a depth of said receptacle, said depth of said receptacle being commensurate with the size of the cells in the fluid medium, whereby when the cells are caused to migrate in the fluid medium through said receptacle, the cells do so in essentially a single layer, the migration of cells from the first end to the second end of said receptacle defining a migration direction for said receptacle;
• b. channeling means for allowing passage of cells through said receptacle in at least one single file;
• c. capping means for covering said receptacle intermediate said first and second ends thereof and for affording visual observation of the migration of the cells through said channeling means and through said receptacle;
• and
• d. means for measuring the amount of energy consumed during passage of the cells through said channeling means.

An apparatus for channelling cells through passageways and simultaneously viewing the cells in a fluid medium, said apparatus comprising:

• a. a substrate having a receptacle located on a side thereof, said receptacle having a floor bound by a pair of upstanding opposing side walls and a first end and a second end, the height of said side walls defining a depth of said receptacle, said depth of said receptacle being commensurate with the size of the cells in the fluid medium, whereby when the cells are caused to migrate in the fluid medium through said receptacle, the cells do so in essentially a single layer, the migration of cells from the first end to the second end of said receptacle defining a migration direction for said receptacle;
• b. channeling means for allowing passage of cells through said receptacle in at least one single file, said channeling means comprising passageways that are curved and are positioned within said receptacle;
• and
• c. capping means for covering said receptacle intermediate said first and second ends thereof and for affording visual observation of the migration of the cells through said channeling means and through said receptacle.

A method for manufacturing a sorting apparatus for fractionating and simultaneously viewing individual microstructures in a fluid medium, said method comprising the steps of:

• a. forming on one side of a substrate a receptacle having a floor bound by a pair of upstanding opposing side walls, the height of said side walls defining a depth of said receptacle, said depth of said receptacle being commensurate with the size of the microstructures in the fluid medium, whereby when the microstructures are caused to migrate in the fluid medium through said receptacle the microstructures do so in essentially a single layer;
• b. building a two-dimensional array of upstanding obstacles in a preselected and reproducible pattern on said floor of said receptacle, the end of each of said obstacles remote from said floor defining the top of said obstacle;
• and
• c. engaging a transparent coverslip to said tops of said obstacles, thereby to preclude migration of the microstructures between said coverslip and said tops of obstacles.

A method for sorting microstructures comprising the steps of:

• a. placing the microstructures into a fluid medium;
• b. introducing the fluid medium with the microstructures therein into a receptacle having a first end and a second end and a floor bound by a pair of upstanding opposing side walls, the height of said side walls defining a depth of said receptacle;
• c. positioning within said receptacle a two-dimensional array of obstacles disposed in a preselected and reproducible pattern;
• and
• d. inducing the microstructures in the fluid to migrate through said array of obstacles positioned within said receptacle.

A method for sorting microstructures, said method comprising the steps of;

• a. placing the microstructures into a fluid medium;
• b. introducing the fluid medium with the microstructures therein into a receptacle having a first end and a second end and a floor bound by a pair of upstanding opposing side walls, the height of said side walls defining a depth of said receptacle;
• c. positioning within said receptacle an array of upstanding obstacles in a preselected and reproducible pattern;
• and
• d. inducing the microstructures in the fluid to migrate through said array of obstacles positioned within said receptacle, said step of inducing comprising the steps of:
• (i) positioning a first electrode on said floor of said receptacle at said first end of said receptacle;
• (ii) positioning a second electrode on said floor of said receptacle at said second end of said receptacle;
• and
• (iii) coupling a power source between said first and second electrodes.

A method for sorting microstructures, said method comprising the steps of:

• a. placing the microstructures into a fluid medium;
• b. introducing the fluid medium with the microstructures therein into a receptacle having a first end and a second end and a floor bound by a pair of upstanding opposing side walls, the height of said side walls defining a depth of said receptacle;
• c. positioning within said receptacle an array of upstanding obstacles in a preselected and reproducible pattern;
• and
• d. inducing the microstructures in the fluid to migrate through said array of obstacles positioned within said receptacle, said step of inducing comprising the step of focusing light onto the microstructures.

A method for sorting microstructures, said method comprising the steps of:

• a. placing the microstructures into a fluid medium;
• b. introducing the fluid medium with the microstructures therein into a receptacle having a first end and a second end and a floor bound by a pair of upstanding opposing side walls, the height of said side walls defining a depth of said receptacle;
• c. positioning within said receptacle an array of upstanding obstacles in a preselected and reproducible pattern;
• and
• d. inducing the microstructures in the fluid to migrate through said array of obstacles positioned within said receptacle, said step of inducing comprising the step of imposing a magnetic field on the microstructures.

A sorting apparatus for fractionating microstructures in a fluid medium, said sorting apparatus comprising:

• a. a substrate with a receptacle formed in a side thereof, said receptacle having a first end, a second end, and a floor bound on opposite sides by a pair of side walls that extend between said first end and second end of said receptacle, migration of the microstructures from said first end of said receptacle to said second end of said receptacle defining a migration direction for said receptacle;
• b. a two-dimensional array of obstacles upstanding from said floor of said receptacle in a preselected and reproducible pattern, the spaces between adjacent of said obstacles in a cross section of said array taken normal to said floor of said receptacle defining pores of a lattice structure capable of interacting with the microstructures to differentially slow the rate of migration of individual types of the microstructures during migration of the microstructures in said migration direction through said array of obstacles, each of said pores of said lattice structure having dimensions comprising:
• i. a height of said pores measured in a direction normal to said floor of said receptacle, said height of said pores of said lattice structure being in a range of from about 0.01 microns to about 20.0 microns;
• and
• ii. a separation distance of said pores measured parallel to said floor of said receptacle between adjacent of said obstacles, said separation distance of said pores of said lattice structure being in a range of from about 0.01 microns to about 50.0 microns;
• and
• c. a coverslip extending across said receptacle from one of said side walls to the other and contacting the end of each of said obstacles opposite from said floor of said receptacle.

A sorting apparatus for fractionating microstructures in a fluid medium, said sorting apparatus comprising:

• a. a substrate having a receptacle located on a side thereof, said receptacle having first and second ends and a floor bound on opposite sides by a pair of upstanding opposed side walls extending between said first and second ends of said receptacle, migration of the microstructures from the first end of said receptacle to said second end of said receptacle defining a migration direction for said receptacle;
• and
• b. sifting means positioned within said receptacle intermediate said first and second ends thereof traversing said migration direction for interacting with the microstructures to partially hinder migration of the microstructures in said migration direction in the fluid medium, said sifting means comprising an array of obstacles upstanding from said floor of said receptacle and arranged in a preselected and reproducible pattern, each of said obstacles having a v-shaped cross section in a plane disposed parallel to said floor of said receptacle, said v-shaped cross section of said obstacles comprising:
• (i) a first arm and
• (ii) a second arm disposed at an angel to said first arm and intersecting said first arm at the vertex of said v-shaped cross section, the ends of said first and second arms remote from said vertex forming the open end of said v-shaped cross section.

A sorting apparatus for fractionating microstructures in a fluid medium, said sorting apparatus comprising:

• a. a substrate having a receptacle located on a side thereof, said receptacle having first and second ends and a floor bound on opposite sides by a pair of upstanding opposed side walls extending between said first and second ends of said receptacle, migration of the microstructures from the first end of said receptacle to said second end of said receptacle defining a migration direction for said receptacle;
• and
• b. sifting means positioned within said receptacle intermediate said first and second ends thereof traversing said migration direction for interacting with the microstructures to partially hinder migration of the microstructures in said migration direction in the fluid medium, said sifting means comprising an array of obstacles upstanding from said floor of said receptacle and arranged in a preselected and reproducible pattern, each of said obstacles having a cup-shaped cross section in a plane disposed parallel to said floor of said receptacle, said cup-shaped cross section of said obstacles comprising:
• (i) a first leg and a second leg positioned in opposed relationship substantially parallel to said migration direction, each of said first and second legs having thereby oppositely disposed first and second ends;
• and
• (ii) a third leg positioned substantially perpendicular to said migration direction and being connected between said first ends of each of said first and second legs, thereby to define between said second ends of said first and second legs the open end of said cup-shaped cross section.