A method is provided for the labeling of individual cells. Labeling is accomplished by coating a particle with at least one dye or nucleic acid sequence encoding a marker protein. The particle is then propelled toward the cell resulting in the particle contacting the cell for a time sufficient for t
A method is provided for the labeling of individual cells. Labeling is accomplished by coating a particle with at least one dye or nucleic acid sequence encoding a marker protein. The particle is then propelled toward the cell resulting in the particle contacting the cell for a time sufficient for the dye or nucleic acid to leave the particle and enter the cell. The present method allows for the differential labeling of individual cells within dense populations of cells.
대표청구항▼
1. A method for individually labeling a cell within a population of cells whereby the cell is differentially labeled relative to neighboring cells within the population, the method comprising propelling a particle coated with a lipophilic hydrophobic dye at the population of cells to cause the parti
1. A method for individually labeling a cell within a population of cells whereby the cell is differentially labeled relative to neighboring cells within the population, the method comprising propelling a particle coated with a lipophilic hydrophobic dye at the population of cells to cause the particle to contact the membrane of the cell, and allowing the dye to diffuse into the cell membrane and thereby differentially label the cell relative to neighboring cells within the population.2. The method of claim 1, wherein the lipophilic hydrophobic dye is a fluorescent dye.3. The method of claim 2, wherein the fluorescent dye is a carbocyanine dye.4. The method of claim 3, wherein the dye is selected from the group consisting of DiO, DiI, DiD, and any combination thereof.5. The method of claim 1, wherein the propelling is by a gas mean.6. The method of claim 5, wherein the gas means is a particle gun.7. The method of claim 1, wherein the particle is a metal particle.8. The method of claim 7, wherein the metal particle is selected from the group consisting of ferrite crystals, gold and tungsten.9. The method of claim 1, wherein the population of cells is part of a tissue.10. The method of claim 9, wherein the tissue is selected from the group consisting of tumor tissue, epidermal tissue, muscle tissue, bone marrow tissue, neural tissue, brain tissue, organ tissue, and human biopsy tissue.11. The method of claim 9, wherein the coated particle is propelled 50-100 μm into the tissue to contact the membrane of the cell.12. The method of claim 11, wherein the coated particle is propelled about 50-70 μm into the tissue.13. The method of claim 1 wherein the labeled cell is a living or a fixed cell.14. A method of imaging a cell, of claim 13, wherein the cell is labeled by the method wherein the labeled cell is a living cell and said cell is imaged within about one minute of the coated particle being propelled at the cell.15. A method of imaging a cell, of claim 13, wherein the cell is labeled by the method wherein the labeled cell is a fixed cell and said cell is imaged within less than about five minutes of the coated particle being propelled at the cell.16. A method of imaging a cell, wherein the cell is labeled by the method of claim 13, wherein the labeled cell is a fixed cell and said cell is imaged within less than about thirty minutes of the coated particle being propelled at the cell.17. The method of claim 1, wherein the cell is a neuron.18. The method of claim 17, wherein the particle contacts an axon.19. The method of claim 17, wherein the particle does not contact the cell body.20. A method for individually labeling cells within a population of cells whereby the cells are differentially labeled relative to neighboring cells within the population, the method comprising propelling a plurality of particles coated with one or more lipophilic hydrophobic dyes at the population of cells to cause the particles to contact the membranes of the cells, and allowing the dye to diffuse into the cell membranes and thereby differentially label the cells relative to neighboring cells within the population.21. The method of claim 20, wherein the particles are coated with more than one lipophilic hydrophobic dye.22. The method of claim 20, wherein one or more dyes the dye is a fluorescent dye.23. The method of claim 20, wherein one or more lipophilic hydrophobic dyes has an emission profile that is distinct from each of the other lipophilic hydrophobic dyes.24. The method of claim 22, wherein the fluorescent dye is a carbocyanine dye.25. The method of claim 24, wherein the carbocyanine dye is selected from the group consisting of DiO, DiI, DiD, and any combination thereof.26. The method of claim 20, wherein the plurality of particles is contained in at least one macroprojectile.27. The method of claim 20, further comprising causing the macroprojectile to contact a macroprojectile stopping means before contacting the cells, the macroprojectile stopping means being capable of stopping the macroprojectile while allowing at least one of the plurality of particles to continue toward the target cell.28. The method of claim 27, wherein the macroprojectile stopping means is a filter.29. The method of claim 28, wherein the filter has a pore size of between about 1 and about 8 μm.30. The method of claim 20, wherein the propelling is by a gas means.31. The method of claim 30, wherein the gas means is a particle gun.32. The method of claim 20, wherein the particles are metal particles.33. The method of claim 32, wherein the metal particles are selected from the group consisting of ferrite crystals, gold and tungsten.34. The method of claim 20, wherein the population or cells is part of a tissue.35. The method of claim 34, wherein the tissue is selected from the group consisting of tumor tissue, epidermal tissue, muscle tissue, bone marrow tissue, neural tissue, brain tissue, organ tissue, and human biopsy tissue.36. The method of claim 34, wherein the coated particles are propelled 50-100 μm into the tissue to contact the membranes of the cells.37. The method of claim 36, wherein the coated particles are propelled about 50-70 μm into the tissue.38. The method of claim 20 wherein the labeled cells are living or fixed cells.39. A method of imaging cells, of claim 38, wherein the cells are labeled by the method wherein the labeled cells are living cells and said cells are imaged within about one minute of the coated particles being propelled at the cell.40. A method of imaging cells, of claim 38, wherein the cells are labeled by the method wherein the labeled cells are fixed cells and said cells are imaged within lees than about five minutes of the coated particles being propelled at the cells.41. A method of imaging cells, wherein the cells are labeled by the method of claim 38, wherein the labeled cells are fixed cells and said cells are imaged within less than about thirty minutes of the coated particles being propelled at the cells.42. The method of claim 20, wherein the cell is a neuron.43. The method of claim 42, wherein the particle contacts an axon.44. The method of claim 42, wherein the particle does not contact the cell body.45. A method for individually labeling cells within a population of cells whereby the cells are differentially labeled relative to neighboring cells within the population, the method comprising propelling a plurality of particles containing a plurality of nucleotide sequences encoding fluorescent proteins having different emission spectra at the population of cells to cause the particles to enter the cells, and allowing expression of the proteins encoded by the nucleotide sequences to occur and thereby differentially label the cells relative to neighboring cells within the population.46. The method of claim 45, wherein the fluorescent proteins with different emission spectra are red fluorescent protein, green fluorescent protein or variants of green fluorescent protein.47. The method of claim 45, wherein the propelling is by a gas means.48. The method of claim 47, wherein the gas means is a particle gun.49. The method of claim 45, wherein the particles are metal particle.50. The method of claim 49, wherein the metal particles axe selected from the group consisting of ferrite crystals, gold and tungsten.51. The method of claim 45, wherein the population of cells is part of a tissue.52. The method of claim 51, wherein the tissue is selected from the group consisting of tumor tissue, epidermal tissue, muscle tissue, bone marrow tissue, neural tissue, brain tissue, organ tissue, and human biopsy tissue.53. The method of claim 51, wherein the coated particles are propelled 50-100 μm into the tissue to enter the cells.54. The method of claim 53, wherein the coated particles are propelled about 50-70 μm into the tissue.
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이 특허에 인용된 특허 (17)
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