What is claimed is:
1. An in vivo method for introducing a therapeutic agent into skin or muscle cells of a subject, said method comprising applying a pulsed electric field to said skin or muscle cells substantially contemporaneously with the application of said therapeutic agent to said skin or muscle cells, such that said therapeutic agent is introduced into said skin or muscle cells.
2. A method according to claim 1, wherein said electric pulse is of sufficient voltage and duration to cause electroporation.
3. A method according to claim 2, wherein, substantially contemporaneously with said application of said electric pulse, a second electric pulse is applied to said skin or muscle cells, said second electric pulse being of sufficient voltage and time to cause transmembrane permealization.
4. A method according to claim 1, wherein said therapeutic agent is selected from a chemotherapeutic agent, a nucleic acid sequence, a polypeptide, and combinations thereof.
5. A method according to claim 4, wherein said nucleic acid sequence is selected from the group consisting of double stranded DNA, single-stranded DNA, complexed DNA, encapsulated DNA, naked RNA, encapsulated RNA, and combinations thereof.
6. A method according to claim 4, said nucleic acid encoding a therapeutic polypeptide, and said nucleic acid sequence being operably associated with a regulatory sequence such that said therapeutic polypeptide is expressed in said skin or muscle cells.
7. A method according to claim 6, wherein said polypeptide comprises an antigen.
8. A method according to claim 6, wherein said polypeptide comprises a pathogen-derived protein, or a peptide hormone.
9. A method according to claim 8, wherein said peptide hormone is selected from the group consisting of calcitonin (CT), parathyroid hormone (PTH), erythropoietin (Epo), insulin, a cytokine, a growth hormone or a growth factor.
10. A method according to claim 6, wherein said polypeptide is a blood coagulation factor or a lymphokine.
11. A method according to claim 10, wherein said lymphokine is selected from the group consisting of tumor necrosis factor, interleukins 1, 2, and 3, lymphotoxin, macrophage activating factor, migration inhibition factor, colony stimulating factor, alpha-interferon, beta-interferon, gamma-interferon and subtypes thereof.
12. A method according to claim 6, wherein said blood coagulation factor is Factor VIII or Factor IX.
13. A method according to claim 6, wherein said nucleic acid sequence further encodes a selectable marker polypeptide.
14. A method according to claim 6, wherein said regulatory sequence comprises a promoter.
15. A method according to claim 14, wherein said promoter is skin or muscle specific.
16. A method according to claim 14, wherein said promoter is selected from the group consisting of CMV, RSV LTR, MPSV LTR, SV40, and keratin specific promoters.
17. A method for inducing an immune response in a subject, said method comprising applying a pulsed electric field to skin or muscle cells of said subject substantially contemporaneously with the application of an immune response-inducing agent to said skin cells or muscle, such that said immune response-inducing agent is introduced into said skin or muscle cells thereby inducing in said subject an immune response.
18. A method according to claim 17, wherein said electric pulse is of sufficient voltage and duration to cause electroporation.
19. A method according to claim 18, wherein, substantially contemporaneously with said application of said electric pulse, a second electric pulse is applied to said skin or muscle cells, said second electric pulse being of sufficient voltage and time to cause iontophoresis.
20. A method according to claim 17, wherein said immune response-inducing agent is selected from a nucleic acid sequence, a polypeptide, and combinations thereof.
21. A method according to claim 20, said nucleic acid encoding a polypeptide, and said nucleic acid sequence being operably associated with a regulatory sequence.
22. A method according to claim 21, wherein said regulatory sequence comprises a promoter.
23. A method according to claim 22, wherein said polypeptide comprises an antigen.
24. A method according to claim 17, wherein said immune response is a humoral response.
25. A method according to claim 17, wherein said immune response is a cellular response.
26. A method according to claim 17, wherein said immune response includes a cellular and a humoral response.
27. A method according to claim 1, wherein said electric pulse comprises a square wave pulse of at least 50 V for about 10 up to 20 ms.
28. A method according to claim 1, wherein said electric pulse is monopolar or bipolar.
29. A method according to claim 17, wherein said electric pulse comprises an exponential decay pulse of 120 V for about 10 up to 20 ms.
30. A method according to claim 17, wherein said electric pulse is monopolar or bipolar.
31. A method according to claim 27, wherein said electric pulse is applied through an electrode selected from the group consisting of a caliper electrode, a meander electrode, a needle electrode, a microneedle array electrode, a micropatch electrode, a ring electrode, and combinations thereof.
32. A method according to claim 31, wherein each electrode of said caliper electrode has an area of about 1 cm2.
33. A method according to claim 31, wherein said caliper electrode is applied to a skin fold having a thickness of about 1 mm to about 6 mm.
34. A method according to claim 29, wherein said electric pulse is applied through an electrode selected from the group consisting of a caliper electrode, a meander electrode, a needle electrode, a microneedle array electrode, a micropatch electrode, a ring electrode, and combinations thereof.
35. A method according to claim 34, wherein each electrode of said caliper electrode has an area of about 1 cm2.
36. A method according to claim 34, wherein said caliper electrode is applied to a skin fold having a thickness of about 1 mm to about 6 mm.
37. A method according to claim 17, wherein said immune response is protective.
38. A method for the therapeutic application of electroporation to skin or muscle cells of a subject for introducing topically applied molecules into said cells, said method comprising:
a) providing an array of electrodes, at least one of the electrodes having a needle configuration for penetrating tissue;
b) inserting the needle electrode into selected tissue;
c) positioning a second electrode of the array of electrodes in conductive relation to the selected tissue; and
d) applying pulses of high amplitude electric signals to the electrodes, proportionate to the distance between the electrodes, for electroporation of said tissue; such that said topically applied molecules are introduced into said skin or muscle cells.
39. A micropatch electrode for use with an electroporation apparatus, said micropatch electrode comprising a substantially planar array of patch elements, each patch element comprising two sets of electrodes, wherein each set of electrodes comprises a first electrode and a second electrode electrically insulated from one another, such that when different electric potentials are applied to said first and second electrodes, a voltage is produced therebetween.
40. A micropatch electrode according to claim 39, wherein said patch elements are separated by electrically insulating material.
41. A micropatch electrode according to claim 40, wherein said electrically insulating material can be pierced by an injection needle.
42. An electrode kit for use in conjunction with electroporation therapy, said kit comprising:
a) a micropatch electrode according to claim 39, and
b) an injection needle, optionally comprising one or more holes disposed along its length and proximal to the needle tip,
wherein said holes are in fluid communication with the hollow interior of said injection needle.
43. An electrode for use with an electroporation apparatus, said electrode comprising:
a) a ring-shaped electrode having an electrically insulating shield, wherein said electrically insulating shield provides support to said ring-shaped electrode and electrically insulates a tissue under treatment employing said electrode, and
b) an electrically conducting injection needle, optionally comprising one or more holes disposed along its length and proximal to the needle tip, wherein said holes are in fluid communication with the hollow interior of said injection needle, wherein a potential difference applied to said ring-shaped electrode and said needle electrode creates a voltage therebetween.
44. An electrode for use with an electroporation apparatus, said electrode comprising an array of a plurality of paired electrode needles and, at least one injection needle, wherein a potential difference applied said paired electrode needles creates a voltage therebetween.
45. An electrode according to claim 44, said injection needle comprising one or more holes disposed along its length and proximal to the needle tip, wherein said holes are in fluid communication with the hollow interior of said needle.
46. An electrode for use with an electroporation apparatus, said electrode comprising a suction generating device comprising a ring electrode disposed about an injection needle electrode, such that when said ring electrode is contacted with skin of a subject and suction is generated by said suction generating device, said skin is pulled up around the injection needle electrode, causing the injection needle electrode to pierce the skin, and
wherein a potential difference applied to said ring-shaped electrode and said injection needle electrode creates a voltage therebetween.
47. An electrode according to claim 46, wherein said suction generating device is a syringe comprising a piston slidably engaged with, and sealingly disposed about said injection needle electrode.
48. A needle electrode for use with an electroporation apparatus, said needle electrode comprising an injection needle and paired electrically conducting wires disposed within the hollow core of said injection needle and protruding from the tip thereof, said electrically conducting wires having an electrically insulated portion and an exposed electrically conducting portion, said exposed electrically conducting portion being at the end of said wires protruding from the tip of said injection needle.
49. A needle electrode for use with an electroporation apparatus, said needle electrode having disposed around a portion of its length a substance-releasing material.
50. A needle electrode according to claim 48, wherein said substance-releasing material is a porous material.
51. A needle electrode for use with an electroporation apparatus, said needle electrode comprising:
a) a hollow central core,
b) at least four holes along its length, wherein said four holes comprise two paired holes, each of said two paired holes comprising a hole proximal to the tip of said needle and a hole distal to said tip of said needle, and
c) a pair of electrically conducting wires having an electrically insulated portion and an exposed electrically conducting portion,
wherein said pair of electrically conducting wires are located in said hollow core, except for said exposed electrically conducting portion which runs outside of said needle, extending outward from said distal paired hole to said proximal paired hole where it re-enters said hollow core.
52. A needle electrode for use with an electroporation apparatus, said electrode comprising a plurality of electrically conducting needles disposed within a depth guide, wherein said tip of said needles extend for a predetermined distance beyond said depth guide.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A method for processing XYZ point cloud of a scene acquired by a GmAPD LADAR, comprising the steps of:
applying low-pass filtering utilizing Deconvolution to the XYZ point cloud to produce a D point cloud; and
displaying an image of the D point cloud.
2. The method as set forth in claim 1, wherein the step of applying low-pass filtering utilizing Deconvolution comprises performing Weiner-Levinson Deconvolution to produce a WLD point cloud and wherein the step of displaying an image of the D point loud comprises displaying an image of the WLD point cloud.
3. The method as set forth in claim 2, wherein the Weiner-Levinson Deconvolution occurs using a Deconvolution Matrix.
4. The method as set forth in claim 3, wherein at least one parameter of Deconvolution Matrix is operator-selectable.
5. The method as set forth in claim 3, wherein the step of displaying an image of the WLD point cloud comprises counting photons at points in the WLD point cloud.
6. The method as set forth in claim 3, further including the step of sharpening the WLD point cloud in the X-Y plane to produce a sharpened point cloud and wherein the step of displaying the image of the WLD point cloud comprises displaying the image of the sharpened point cloud.
7. The method as set forth in claim 6, wherein the step of sharpening the WLD point cloud in the X-Y plane to produce the sharpened point cloud comprises highpass filtering.
8. The method as set forth in claim 3, further including the step of mitigating timing uncertainty in the WLD point cloud by deconvolution to produce a deconvolved point cloud and wherein the step of displaying an image of the WLD point cloud comprising displaying and image of the deconvolved point cloud.
9. The method as set forth in claim 8, wherein the step of mitigating timing uncertainty in the WLD point cloud by deconvolution comprises deconvoluting the WLD point cloud in the vertical direction.
10. The method as set forth in claim 8, further including the step of thresholding the sharpened point cloud to produce a thresholded point cloud and wherein the step of mitigating the timing uncertainty in the WLD point cloud by deconvolution comprises mitigating the timing uncertainty in the thresholded point cloud.
11. The method as set forth in claim 8, further including the step of Z-clipping the XYZ point cloud to produce a Z-clipped point cloud and wherein the step of performing low-pass filtering utilizint Deconvolution on XYZ point cloud comprises performing low-pass tiltering utilizing Deconvolution on the Z-clipped point cloud.
12. The method as set forth in claim 11, wherein the step of Z-clipping the XYZ point cloud comprises adaptive histogramming.
13. The method as set forth in claim 6, further including the step of thresholding the WLD point cloud to produce a thresholded point cloud and wherein the step of sharpening the WLD point cloud comprises sharpening the thresholded point cloud.
14. The method as set forth in claim 9, further including the step of thresholding and cleansing the deconvolved point cloud in the vertical direction to produce a thresholdedcleansed point cloud and wherein the step of displaying an image of the deconvolved point cloud comprises displaying an image of the thresholdedcleansed point cloud.
15. A method for processing a XYZ point cloud of a scene acquired by a GmAPD LADAR, comprising the steps of:
Z-clipping the XYZ point cloud adaptive histogramming to produce a Z-clipped point cloud;
applying low-pass filtering utilizing Weiner-Levinson Deconvolution to the XYZ point cloud, utilizing a Deconvolution Matrix having at least one parameter that is operator-selectable to produce a WLD point cloud;
thresholding the WSD point cloud to produce a first thresholded point cloud;
sharpening the WLD point cloud in the X-Y plane by highpass filtering to produce a sharpened point cloud;
thresholding the sharpened point cloud to produce a second thresholded point cloud;
mitigating timing uncertainty in the second thresholded point cloud by deconvolving the second thresholded point cloud in the vertical direction to produce a deconvolved point cloud;
thresholding and cleansing the deconvolved point cloud in the vertical direction to produce a thresholdedcleansed point cloud; and
displaying an image of the thresholdedcleansed point cloud by counting photons at points in the thresholdedcleansed point cloud.
16. A system for processing a XYZ point cloud of a scene acquired by a GmAPD LADAR, comprising in combination:
an image processor that performs low-pass filtering utilizing Deconvolution to the XYZ point cloud to produce a D point cloud; and
a display for displaying an image of the D point cloud.
17. The system as set forth in claim 16, wherein the image processor applies the low-pass filtering utilizing Deconvolution using Weiner-Levinson Deconvolution to produce a WLD point cloud and wherein the display displays an image of the WLD point cloud.
18. The system as set forth in claim 17, wherein the image processor performs said Weiner-Levinson Deconvolution using a Deconvolution Matrix.
19. The system as set forth in claim 18, wherein at least one parameter of said Deconvolution Matrix is operator-selectable.
20. The system as set forth in claim 18, wherein the image processor counts photons at points in the WLD point cloud for display.