Needle array and method of introducing biological substances into living cells using the needle array

• US 5,457,041 A
• Filed: 03/25/1994
• Issued: 10/10/1995
• Est. Priority Date: 03/25/1994
• Status: Expired due to Term

• Current Assignee: Leidos, Inc. (Leidos Holdings, Inc.)
• Original Assignee: Science Applications International Corporation
• Inventors: Ginaven, Robert O., Facciotti, Daniel
• Primary Examiner(s): Beisner, William H.
• Application Number:
  US08/218,059
• Time in Patent Office: 564 Days
• Field of Search: 435/172.1, 435/172.3, 435/287, 435/292, 435/293, 935/52, 935/53, 935/85, 422/100
• US Class Current: 435/455
• CPC Class Codes: A61M 2037/0023 Drug applicators using microneedles
• CPC Class Codes: A61M 37/0015 by using microneedles
• CPC Class Codes: C12M 35/00 Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion machines for extracting juice from animal or plant tissue by electroplasmolysis A23N1/006, processes employing electric or wave energy B01J19/08; treatment of microorganisms or enzymes with electrical or wave energy C12N13/00; methods for cell fusion C12N15/02; introduction of foreign genetic material C12N15/87
• CPC Class Codes: C12N 15/8207 by mechanical means, e.g. microinjection, particle bombardment, silicon whiskers

Abstract

An array of micro-needles extending from a support substrate and having tip portions shaped and dimensioned to (1) carry a biological substance and (2) pierce the surface of a target tissue and penetrate target cells within the tissue where the biological substance transfers from the tip portions and deposits within the target cells.

Claims

1. Apparatus for depositing biological substances into target cells in living plant or animal target tissue, the apparatus comprising:

a mechanical holder for securing a target tissue;

a target tissue secured by the holder and including target cells having a known width and depth from a surface of the target tissue;

an array of micro-needles substantially normal to and extending from a surface of a support substrate for piercing the surface of the target tissue to penetrate the target cells, the needles having a length greater than the depth of the target cells and sufficient to pierce the target cells when the needles are forced into the tissue with the surface of the support substrate adjacent to the surface of the tissue, and a spacing substantially equal to the width of the target cells which allows each needle to pierce one of the target cells, at least a portion of each needle carrying a biological substance for transfer to and deposit within the target cells upon a piercing thereof by the needles;

means for supporting the array of micro-needles relative to the holder; and

means for moving the array of micro-needles relative to the holder to penetrate the target tissue until the surface of the support substrate is adjacent the surface of the target tissue.

2. The apparatus of claim 1 wherein the needles have a base width at the surface of the support substrate of between about 0.5 and 3 microns.

3. The apparatus of claim 2 wherein the length of the needles is between about 10 and 25 microns.

4. The apparatus of claim 3 wherein the spacing of the needles is between about 5 and about 20 microns.

5. The apparatus of claim 4 wherein the needle array has a size up to about 20 needles by 20 needles.

6. The apparatus of claim 1 wherein the needles include tip portions at least some of which are pointed.

7. The apparatus of claim 1 wherein the needles include tip portions at least some which are concave to define a biological substance receiving cavities.

8. The apparatus of claim 7 wherein the biological substance is carried by micro-particles in the cavities.

9. The apparatus of claim 1 wherein the needles include tip portions at least some of which include a major pointed tissue piercing peak surrounded by minor pointed peaks for carrying the biological substance in cavities between the major and minor peaks.

10. The apparatus of claim 1 wherein the needles include tip portions at least some of which are of a ragged, cage-like construction including a plurality of pointed peaks having biological substance receiving cavities therebetween.

11. The apparatus of claim 1 wherein;

the mechanical holder is movable in X and Y directions relative to the means for supporting the array of micro-needles, and

the means for supporting the array of micro-needles comprises a tubular holder, a movable slide in the tubular holder, and means for securing the array of micro-needles at one end of the slide to move relative to the target tissue on the mechanical holder.

12. The apparatus of claim 11 further including means at an opposite end of the slide for moving the slide and array of micro-needles to pierce the target tissue.

13. The apparatus of claim 11 further including a microscope for simultaneously viewing the array of micro-needles and the target tissue to allow for controlled X and Y movement of the mechanical holder relative to the tubular holder to align the target tissue and the array of micro-needles.

14. The apparatus of claim 13 wherein:

The microscope including a viewing lens is spaced from the target tissue and array of micro-needles, and

the apparatus further comprises an optics system including beam splitter means between the array of micro-needles and the target tissue for simultaneously reflecting images of the array of micro-needles and the target tissue to the microscope for simultaneous viewing by an operator wherein the beam splitter means is mounted for movement from the optic system after the array of micro-needles and target tissue have been aligned.

15. The apparatus of claim 13 wherein the microscope includes an object viewing lens aligned with the array of micro-needles and target tissue for a simultaneous viewing thereof.

16. A method of depositing a biological substance into target cells in a living plant or animal tissue, the target cells having a known width and depth from a surface of the target tissue, the method comprising the step of:

(a) securing the tissue on a base;

(b) mounting an array of micro-needles having tip portions carrying the biological substance or biological substance carrying micro-particles to a holder and aligning the array of needles with the tissue on the base, the needles having a length greater than the depth of the target cells and sufficient to pierce the target cells when the needles are forced into the tissue with the surface of a support substrate for the micro-needles adjacent to the surface of the tissue, and a spacing substantially equal to the width of the target cells to allow each needle to pierce one of the target cells;

(c) moving the holder relative to the tissue to force the array of needles into the tissue until the tip portions penetrate the target cells to allow the biological substance to transfer into the target cells; and

(d) withdrawing the needle away from the tissue.

17. The method of claim 16 wherein aligning the array of needles and the tissue comprises simultaneously viewing the array and tissue with a microscope and while so viewing, moving the base relative to the holder until the array and tissue are aligned.

18. The method of claim 16 further comprising breaking off tip portions of the needles carrying the biological substance within the target cell to remain therein upon the withdrawing of the needles from the tissue.