Methods tip assemblies and kits for introducing material into cells
Methods, tip assemblies and kits are provided for introducing material into cells. The tip assemblies include an attachment portion, a channel portion, and a constriction that function to reduce fluid pressure as a fluid passes through the constriction portion from the channel portion, whereby the tip assemblies form pores in the membranes of cells and introduce material into the cells. The material includes for example one selected from the group of: an inorganic compound, a drug, a genetic material, a protein, a carbohydrate, a synthetic polymer, and a pharmaceutical composition.
1. A tip assembly for introducing a composition in a fluid into cells comprising:
an attachment portion open to the atmosphere and proximally fitting a flow device that generates at least one of a positive pressure and a negative pressure for directing the fluid to flow through the tip assembly;
a channel portion contiguous to and distal to the attachment portion and the flow device;
a constriction portion contiguous with the channel portion, wherein a constriction portion inner diameter and a constriction portion cross sectional area are smaller than a channel portion inner diameter and a channel portion cross sectional area, wherein a distal end of the constriction portion comprises an opening for ejecting or drawing the fluid, wherein increased fluid velocity and a decreased pressure in the fluid in the constriction portion compared to velocity and pressure in the channel portion enhances formation of membrane pores in the cells, whereby the tip assembly introduces the composition into the cells through the membrane pores, wherein the channel portion curvature and constriction inner curvature characterize a fluid path for the fluid flowing through the tip assembly, wherein the channel portion curvature and constriction inner curvature necessary for introducing the composition into the cells comprises a formula represented in two dimensions is
f ( x ) =p 1 x 7 +p 2 x 6 +p 3 x 5 +p 4 x 4 +p 5 x 3 +p 6 x 2 +p 7 x +p 8
wherein x is a radial distance from a center axis on the fluid path to an inner surface of the tip and coefficients with 95% confidence bounds in parentheses comprise: p1 is −2.611e −16 (−6.043e −16 , 8.206e −17 ), p2 is 3.954 e−13 (−3.195 e−13 , 1.11 e−12 ), p3 is −1.845 e−10 (−7.821 e−10 ,4.131 e−10 ), p4 is 1.662 e−08 (−2.394 e−07 , 2.726 e−07 ), p5 is 7 . 537 e−06 (−5.186 e−05 , 6.694 e−05 ), p6 is −0.002137 (−0.009375, 0.005101), p7 is −0.003185 (−0.4114, 0.4051), and p8 =268.6 (261, 276.3), or
f ( x ) =p 1 x 8 +p 2 x 7 +p 3 x 6 +p 4 x 5 +p 5 x 4 +p 6 x 3 +p 7 x 2 +p 8 x +p 9
wherein x is a radial distance from a center axis on the fluid path to an inner surface of the tip and coefficients with 95% confidence bounds in parentheses comprise: p1 is 2.285 (1.388, 3.182), p2 is 3.465 (1.782, 5.149), p3 is −15.68 (−20.04, −11.32); p4 is −20.38 (−27.24, −13.52); p5 is 44.27 (36.5, 52.04); p6 is 53.96 (45.69, 62.23); p7 is −58.72 (−64.02, −53.42), p8 is −123.5 (−126.4, −120.6), and p9 is 186.5 (185.6, 187.4).
2. The tip assembly according to claim 1 , wherein the tip assembly is characterized in that it is at least selected from: disposable, modular, reusable, transparent, and translucent.
3. The tip assembly according to claim 1 , wherein at least one of the attachment portion, the channel portion, and the constriction portion comprises a substance selected from the group of: a glass, a metal, a plastic, a polymer, a nano-based composition, and a composite material comprising at least two different types of substances.
4. The tip assembly according to claim 1 , wherein an inner diameter or outer diameter of the attachment portion of the tip assembly fits the flow device in a male to female arrangement respectively.
5. The tip assembly according to claim 1 , wherein the channel portion cross sectional area or the constriction portion cross sectional area is bounded by a circle, an ellipse, a rectangle or a square, and wherein the distal end of the constriction portion comprises an inner diameter that is less than or substantially equal to an inner diameter of the opening.
6. The tip assembly according to claim 1 , wherein the tip assembly contains a volume which is: about 2 microliters (μl), about 20 μl, about 50 μl, about 200 μl, about 400μl, about 500 μl, about 1 milliliter (ml), about 5 ml, and about 10 ml, and wherein the channel portion inner diameter is about 1.0 millimeter (mm) to about 10.0 mm, and the diameter of the constriction inner diameter is about 0.05 mm to about 2.0 mm, wherein the constriction inner diameter is smaller than the channel portion inner diameter.
7. The tip assembly according to claim 1 , further comprising a shoulder extending laterally from of an outward surface of the attachment portion, and a lower ejector section of the flow device is operative to remove the tip assembly from the flow device.