CA1302753C - Non-destructive optical trap for biological particles and method of doing same - Google Patents
Non-destructive optical trap for biological particles and method of doing sameInfo
- Publication number
- CA1302753C CA1302753C CA000577697A CA577697A CA1302753C CA 1302753 C CA1302753 C CA 1302753C CA 000577697 A CA000577697 A CA 000577697A CA 577697 A CA577697 A CA 577697A CA 1302753 C CA1302753 C CA 1302753C
- Authority
- CA
- Canada
- Prior art keywords
- trap
- particles
- light beam
- optical trap
- biological particles
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
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Classifications
-
- H—ELECTRICITY
- H05—ELECTRIC TECHNIQUES NOT OTHERWISE PROVIDED FOR
- H05H—PLASMA TECHNIQUE; PRODUCTION OF ACCELERATED ELECTRICALLY-CHARGED PARTICLES OR OF NEUTRONS; PRODUCTION OR ACCELERATION OF NEUTRAL MOLECULAR OR ATOMIC BEAMS
- H05H3/00—Production or acceleration of neutral particle beams, e.g. molecular or atomic beams
- H05H3/04—Acceleration by electromagnetic wave pressure
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D43/00—Separating particles from liquids, or liquids from solids, otherwise than by sedimentation or filtration
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/32—Micromanipulators structurally combined with microscopes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
Abstract
NON-DESTRUCTIVE OPTICAL TRAP FOR BIOLOGICAL
PARTICLES AND METHOD OF DOING SAME
Biological particles are successfully trapped in a single-beam gradient force trap using an infrared laser. The high numerical aperture lens objective in the trap is also used for simultaneous viewing.
Several modes of trapping operation are presented. (FIG. 1)
PARTICLES AND METHOD OF DOING SAME
Biological particles are successfully trapped in a single-beam gradient force trap using an infrared laser. The high numerical aperture lens objective in the trap is also used for simultaneous viewing.
Several modes of trapping operation are presented. (FIG. 1)
Description
~3(~Z~S;~
NON-DESTRUCTIVE OPTICAL TRAP FOR BIOLOGICAL
PARTICLES AND METHOD OF DOING SAME
Tech~,l ~ield This invention relates to trapping of particles using a single-beam 5 gradient force trap.
Rackg~ound Q~ tl~ In~Qa Single-beam gradient force traps have been demonstrated for neutral atoms and dielectric particles. Generally, the single-beam gradient force trap consists only of a strongly focused laser beam having an approximately Gaussian 10 transverse intensity profile. In these traps, radiation pressure scattering and gradient force components are combined to give a point of stable equilibrium located close to the focus of the laser beam. Scattering force is proportional to optical intensity and acts in the direction of the incident laser light. Gradient force is proportional to the optical intensity gradient and points in the direction 15 of the intensity gradient.
Particles in a single-beam gradient force trap are confined transverse to the laser beam axis by a radial component of the gradient force. Stabilizing theparticle along the axial direction of the trap is achieved by strongly focusing the laser beam to have the axial component of gradient force dominate the 20 scattering force in the trap region.
In prior work using single-beam gradient force optical traps on dielectric particles, trapping was demonstrated with a visible light laser source (A = 514.5 nm.) focused by a high numerical aperture lens objective. See A. Ashkin et al., ~ti~ T ektec~, Vol. 11, p 288-~0. The dielectric particles were closely spherical 25 or spheroidal in shape and ranged in size from 10,um diameter Mie glass spheres (a! > > )~) down to 200 Angstrom diameter Rayleigh particles (Cl! < < ~). Use ofsuch regularly shaped particles in the Mie regime was desirable as taught in this and other articles.
For Mie particles, both the magnitude and direction of the forces depend 30 on the particle shape. This restricts trapping to fairly simple shapes such as spheres, ellipsoids, or particles whose optical scattering varies slowly with orientation in the trap. In the Rayleigh regime, the particle acts as a dipole and the direction of force is independent of particle shape; only the magnitude of ' ' . . . . ..
:,.. .. . . . .
'' ' ' .
`-` 13~Z753 force varies with particle orientation.
It is not an insignificant result of the prior work that silica and other dielectric particles experienced varying amounts of irreversible optical damage ~rom the trap. While it was suggested that the single-beam trap and the prior results would be extensible to S biological particles, the resulting damage from exposure in the trap would destroy or significantly incapacitate the biological particles and render them useless. Also, since prior optical traps have been defined for quite regular-shaped, dielectric particles, their extension to biological particles is cast in doubt because regularity of shape is not an attribute of biological particles.
10 SummarY of the Invention Biological particles are successfully trapped in a single-beam gradient force optical trap incorporating an infrared light source. Reproduction of trapped particles has been observed. After release from the trap, particles exhibit normal motility and continued reproductivity even after trapping for several life cycles at a high laser power of 160 mW.
In one embodiment, the high numerical aperture lens objective in the single-beamgradient force trap is used for simultaneous viewing of the trapped biological particles.
Two single-beam gradient force optical traps are introduced into the same cell to permit three-dimensional manipulation of the biological particles.
In accordance with one aspect of the invention there is provided apparatus for 20 generating a single-beam gradient force optical trap of particles, said apparatus comprising a laser for generating a light beam at a predetermined wavelength and means for focusing said light beam with sufficient convergence to form said optical trap in a predetermined region, said apparatus characterized in that said predetermined wavelength is substantially included in the infrared range of wavelengths between 0.8 ,um and 1.8 ,um inclusively, so 25 that said trap non-destructively confines at least one biological particle.
In accordance with another aspect of the invention there is provided apparatus for generating a single-beam gradient force optical trap of particles, said apparatus being comprised of a laser for generating a light beam at a predetermined wavelength and means for focusing said light beam with sufficient convergence to form said optical trap in 30 a predeterrnined region, said apparatus characterized in that said predetermined 13~t;27S3 2a wavelength is substantially included in the infrared range of wavelengths, so that said trap non-destructively confines at least one biological particle, said apparatus further including means for generating a second light beam substantially at the predetermined wavelength, 5 said second light beam focused by said focusing means to form a second optical trap in a second predetermined region.
Brief Description of the Drawin .
A more complete understanding of the invention may be obtained by reading the following description of a specific illustrative embodiment of the invention in conjunction 10 with the appended drawing in which:
FIG. 1 is a cross-sectional schematic diagram of an embodiment of the invention;FIG. 2 is a cross-sectional schematic diagram of an embodiment of the invention employing two single-beam gradient force traps in one cell; and FIGS. 3 th~ough 5 are schematic drawings of different modes oE operation for an 15 optical trap on particles in a cell.
,~, .. ~.. ...
.
~3~7$;;~
Detailed I)escription Single-beam gradient force optical traps are useful for confining, isolating, translating and manipulating at least one particle in a group of particles enclosed in a cell or hanging droplet or the like. Special problems 5 surface when the particles are biological. For example, absorption of the optical energy in the trap by the confined particle may lead to particle annihilation or a significant loss of particle motility. Also, as the wavelength of the light beam is varied to avoid the aforementioned problem, the intensity of the optical trap may be sufficiently decreased so as to be rendered ineffective for the particles of 10 interest. While the wavelength selected may be sufficient for effective operation of the optical trap, it may be at a wavelength which is absorbed by the medium surrounding the particles and, therefore, which leads to heat generation within the cell. Clearly, many factors must be considered when selecting the operating wavelength for the optical trap.
In the prior optical trap experiments reported in the literature, particle sensitivity has not been an issue. This is generally attributed to the fact thatdielectric particles have homogeneous compositions and uniformly regular shapes 90 that it is straightforward to observe the effect of the trap on one particle or portion of a particle and accurately predict the effect on other 20 particles or on other portions of the same dielectric particle. For biological particles, sensitivity of the particles is extremely important. Biological particles have heterogeneous compositions and irregular shapes. Hence, the effect of the trap on one part of a biological particle i9 in no way determinative of the effect in another portion of the same particle.
FIG. 1 shows a cross-sectional schematic diagram of apparatus for creating a single-beam gradient force optical trap in accordance with the principles of this invention. IR laser 10 is a standard laser emitting a coherent light beam substantially in the infrared range of wavelengths, for example, 0.8 ~m to 1.8 ~m.
Light beam 11 from IR laser 10 impinges upon a combination of optics elements for focusing the light beam with a sufficient degree of convergence to form a single-beam gradient force optical trap for confining biological particles at a desired position. The combination of optics elements includes an adjustably mounted diverging lens 12 and a high convergence lens 23.
.. ~.. .
13~2753 Lens 12 is adjustable in any of three dimensions (x, y, z) by manipulating adjustable mount 13. It is important that lens 12 expand the spot size of light beam 11 to cover a substantial area on the surface of lens 23. As shown in FIG. 1, diverging light beam 14 impinges on a large portion of the facing surface 5 of lens 23 so that relatively high intensity of beam 14 fills the aperture of lens 23. In order to create the forces required for operation of the single-beam gradient force optical trap, it is desirable that lens 23 be capable of focusing to a spot size less than ~\ approaching ~/2. In an example from experimental practice, lens 23 is a strong or high convergence water immersion microscope 10 objective lens having a numerical aperture of approximately 1.25 (measured inwater). wherein the numerical aperture is defined as the refractive index for the medium multiplied by the sine of the half angle covered by the converging light beam. Element 24 depicts the liquid (water or oil) in which lens 23 is immersed for improved optical coupling into cell 25.
The optical trap is shown within cell 25 with particle 27 captured in the trap. Particle 27 is suspended in a liquid medium such as water, for example, which is enclosed by cell 25. Cell 25 is a transparent enclosure for enclosing the suspended biological particles or a transparent slide from which particle containing droplets can be hung. In one example, cell 25 has dimensions of 20 1 cm. x 3 cm. x 100 ,~m.
The position of cell 25 is adjustable in three dimensions (x, y, z) by the use of adjustable mount 2B. In practice, mount 26 is useful in locating and manipulating the biological particles.
Viewing of biological particles in the trap is accomplished directly or 25 through the use of a monitor. While other types of viewing such as viewing directly in cell 25 are possible, it is an added feature of the present invention the viewing is accomplished through the same lens objective which simultaneously creates the optical trap.
Illumination for viewing is provided by visible light source 2~ and is 30 projected through converging lens 28 onto the particles in the field of view.High resolution viewing occurs with the aid of lens 23 through which the visiblelight passes toward either the eyepiece or the monitor 18. For direct viewing, visible light shown as a dashed line is reflected from beam splitter 1~ to microscope eyepiece 21. Infrared blocking filter 22 is placed in front of eyepiece , ,, ... ~ ... .. .
:~3~ 7'S3 21 to isolate the viewing optics (viewer's eye) from back reflections from cell 25.
For monitoring, the visible light passes through beam splitter 1~ and is reflected from beam splitter 15 toward infrared blocking filter 17 and finally monitor 18.Infrared blocking rllter 17 isolates the monitor from back reflections from cell5 25.
In FIG. 2, the apparatus shown in FIG. 1 is augmented by a second infrared laser source and optics to create a second single-beam gradient force optical trap in cell 25. Infrared laser source 30 generates light beam 31 impinging on adjustably mounted diverging lens 32. Lens 32 causes beam 31 to 10 emerge in a diverging pattern as light beam 34. Adjustment of lens 32 is accomplished in three dimensions (x, y, z) via adjustable mount 33. Light beam 34 is reflected by mirror 35 which coincidently permits transmission of light beam 14. This would occur by judiciously choosing different wavelengths of operation for the separate laser sources. On the other hand, element 35 can be 15 realized as a beam splitter which would reflect approximately half of the light beam incident thereon and transmit the remaining half. As shown in FIG. 2, light beam 34 is converged by lens 23 to form a second trap in cell 25. Particle36 i9 confned in the second trap.
While not shown, it should now be apparent to those skilled in the art 20 that a second trap may be created in the cell by utilizing an additional set of optics including another high convergence microscope. The second trap may be created from light entering the cell on the side opposite the beam for the firsttrap or, for that matter, at any angle to the beam for the first trap.
Manipulation or orientation of particles is achieved by grabbing each end 2S of a rod-like particle, for example, and moving it at will.
In operation, it is necessary to move the trapped biological particles into the viewing plane. This is carried out by adjusting the position of the diverging lens or lenses. Similarly, translation, separation or isolation of the biological particles is easily affected by adjusting mount 26 by the desired amount.
FIGs. 3 through 5 show several modes of operation for the same optical trap. FIG. 3 shows the conventional mode of operation in which the focus of the beam from lens 23 lies within cell 25 and the trapping action relies on the backward gradient component of the optical force. Depending on the size of the particles, it is possible to trap up to approximately four or five particles within ~Z~
the trap at one time.
Both modes shown in FIGs. 4 and 5 require less intensity than for the trap in FIG. 3. In FIG. 4, the bottom plate of cell 25 provides the backward trapping force and the gradient provides the transverse trapping force. It is 5 possible to trap approximately twelve or more biological particles at one time.
In FIG. 5, the scattering force of the focused light beam provides transverse confinement due to its inward direction; backward trapping i9 supplied by the bottom plate of cell 25. In the latter mode of operation, it is possible to trapsignificantly greater numbers of particles than for the modes shown in FIGs. 3 10 and 4.
Various biological particles have been isolated, confined and transported in this type of optical trap. For example, some biological particles successful trapped are tobacco mosaic viruses (See Ashkin et al., Science, Vol. 235, pp.
1517-20 (1~87).), yeast, E. coli bacteria, blood cells containing hemoglobbin, and 15 complex cells or parts of cells containing chlorophyll structures.
In general, the biological particles investigate do not have the regular shape of the dielectric spheres studied earlier. For example, passive, string-like organisms were trapped wherein the organism was approximately 50 ,um long and approximately 1 ~m in diameter. In the case of tobacco mosaic virus, the 20 particles resemble a cylinder about 200 angstroms in diameter and 3100 angstroms long.
It is a significant attribute of the present invention that particle motility is preserved and reproductivity of the particles is maintained. Reproduction by trapped biological particles has been observed with offspring remaining in the 25 trap. In other words, the optical trap permits non-destructive manipulation of biological particles at optical powers approaching several hundred milliwatts.
It should be noted that the use of infrared light results in a lower intensity trap at the focal spot for the same laser power than for traps using visible light. However, the forces in the trap are approximately equal. Thus, 30 the infrared trap has the added benefit over visible light traps of inducing less local heating in the focal spot.
^
...... .. .
NON-DESTRUCTIVE OPTICAL TRAP FOR BIOLOGICAL
PARTICLES AND METHOD OF DOING SAME
Tech~,l ~ield This invention relates to trapping of particles using a single-beam 5 gradient force trap.
Rackg~ound Q~ tl~ In~Qa Single-beam gradient force traps have been demonstrated for neutral atoms and dielectric particles. Generally, the single-beam gradient force trap consists only of a strongly focused laser beam having an approximately Gaussian 10 transverse intensity profile. In these traps, radiation pressure scattering and gradient force components are combined to give a point of stable equilibrium located close to the focus of the laser beam. Scattering force is proportional to optical intensity and acts in the direction of the incident laser light. Gradient force is proportional to the optical intensity gradient and points in the direction 15 of the intensity gradient.
Particles in a single-beam gradient force trap are confined transverse to the laser beam axis by a radial component of the gradient force. Stabilizing theparticle along the axial direction of the trap is achieved by strongly focusing the laser beam to have the axial component of gradient force dominate the 20 scattering force in the trap region.
In prior work using single-beam gradient force optical traps on dielectric particles, trapping was demonstrated with a visible light laser source (A = 514.5 nm.) focused by a high numerical aperture lens objective. See A. Ashkin et al., ~ti~ T ektec~, Vol. 11, p 288-~0. The dielectric particles were closely spherical 25 or spheroidal in shape and ranged in size from 10,um diameter Mie glass spheres (a! > > )~) down to 200 Angstrom diameter Rayleigh particles (Cl! < < ~). Use ofsuch regularly shaped particles in the Mie regime was desirable as taught in this and other articles.
For Mie particles, both the magnitude and direction of the forces depend 30 on the particle shape. This restricts trapping to fairly simple shapes such as spheres, ellipsoids, or particles whose optical scattering varies slowly with orientation in the trap. In the Rayleigh regime, the particle acts as a dipole and the direction of force is independent of particle shape; only the magnitude of ' ' . . . . ..
:,.. .. . . . .
'' ' ' .
`-` 13~Z753 force varies with particle orientation.
It is not an insignificant result of the prior work that silica and other dielectric particles experienced varying amounts of irreversible optical damage ~rom the trap. While it was suggested that the single-beam trap and the prior results would be extensible to S biological particles, the resulting damage from exposure in the trap would destroy or significantly incapacitate the biological particles and render them useless. Also, since prior optical traps have been defined for quite regular-shaped, dielectric particles, their extension to biological particles is cast in doubt because regularity of shape is not an attribute of biological particles.
10 SummarY of the Invention Biological particles are successfully trapped in a single-beam gradient force optical trap incorporating an infrared light source. Reproduction of trapped particles has been observed. After release from the trap, particles exhibit normal motility and continued reproductivity even after trapping for several life cycles at a high laser power of 160 mW.
In one embodiment, the high numerical aperture lens objective in the single-beamgradient force trap is used for simultaneous viewing of the trapped biological particles.
Two single-beam gradient force optical traps are introduced into the same cell to permit three-dimensional manipulation of the biological particles.
In accordance with one aspect of the invention there is provided apparatus for 20 generating a single-beam gradient force optical trap of particles, said apparatus comprising a laser for generating a light beam at a predetermined wavelength and means for focusing said light beam with sufficient convergence to form said optical trap in a predetermined region, said apparatus characterized in that said predetermined wavelength is substantially included in the infrared range of wavelengths between 0.8 ,um and 1.8 ,um inclusively, so 25 that said trap non-destructively confines at least one biological particle.
In accordance with another aspect of the invention there is provided apparatus for generating a single-beam gradient force optical trap of particles, said apparatus being comprised of a laser for generating a light beam at a predetermined wavelength and means for focusing said light beam with sufficient convergence to form said optical trap in 30 a predeterrnined region, said apparatus characterized in that said predetermined 13~t;27S3 2a wavelength is substantially included in the infrared range of wavelengths, so that said trap non-destructively confines at least one biological particle, said apparatus further including means for generating a second light beam substantially at the predetermined wavelength, 5 said second light beam focused by said focusing means to form a second optical trap in a second predetermined region.
Brief Description of the Drawin .
A more complete understanding of the invention may be obtained by reading the following description of a specific illustrative embodiment of the invention in conjunction 10 with the appended drawing in which:
FIG. 1 is a cross-sectional schematic diagram of an embodiment of the invention;FIG. 2 is a cross-sectional schematic diagram of an embodiment of the invention employing two single-beam gradient force traps in one cell; and FIGS. 3 th~ough 5 are schematic drawings of different modes oE operation for an 15 optical trap on particles in a cell.
,~, .. ~.. ...
.
~3~7$;;~
Detailed I)escription Single-beam gradient force optical traps are useful for confining, isolating, translating and manipulating at least one particle in a group of particles enclosed in a cell or hanging droplet or the like. Special problems 5 surface when the particles are biological. For example, absorption of the optical energy in the trap by the confined particle may lead to particle annihilation or a significant loss of particle motility. Also, as the wavelength of the light beam is varied to avoid the aforementioned problem, the intensity of the optical trap may be sufficiently decreased so as to be rendered ineffective for the particles of 10 interest. While the wavelength selected may be sufficient for effective operation of the optical trap, it may be at a wavelength which is absorbed by the medium surrounding the particles and, therefore, which leads to heat generation within the cell. Clearly, many factors must be considered when selecting the operating wavelength for the optical trap.
In the prior optical trap experiments reported in the literature, particle sensitivity has not been an issue. This is generally attributed to the fact thatdielectric particles have homogeneous compositions and uniformly regular shapes 90 that it is straightforward to observe the effect of the trap on one particle or portion of a particle and accurately predict the effect on other 20 particles or on other portions of the same dielectric particle. For biological particles, sensitivity of the particles is extremely important. Biological particles have heterogeneous compositions and irregular shapes. Hence, the effect of the trap on one part of a biological particle i9 in no way determinative of the effect in another portion of the same particle.
FIG. 1 shows a cross-sectional schematic diagram of apparatus for creating a single-beam gradient force optical trap in accordance with the principles of this invention. IR laser 10 is a standard laser emitting a coherent light beam substantially in the infrared range of wavelengths, for example, 0.8 ~m to 1.8 ~m.
Light beam 11 from IR laser 10 impinges upon a combination of optics elements for focusing the light beam with a sufficient degree of convergence to form a single-beam gradient force optical trap for confining biological particles at a desired position. The combination of optics elements includes an adjustably mounted diverging lens 12 and a high convergence lens 23.
.. ~.. .
13~2753 Lens 12 is adjustable in any of three dimensions (x, y, z) by manipulating adjustable mount 13. It is important that lens 12 expand the spot size of light beam 11 to cover a substantial area on the surface of lens 23. As shown in FIG. 1, diverging light beam 14 impinges on a large portion of the facing surface 5 of lens 23 so that relatively high intensity of beam 14 fills the aperture of lens 23. In order to create the forces required for operation of the single-beam gradient force optical trap, it is desirable that lens 23 be capable of focusing to a spot size less than ~\ approaching ~/2. In an example from experimental practice, lens 23 is a strong or high convergence water immersion microscope 10 objective lens having a numerical aperture of approximately 1.25 (measured inwater). wherein the numerical aperture is defined as the refractive index for the medium multiplied by the sine of the half angle covered by the converging light beam. Element 24 depicts the liquid (water or oil) in which lens 23 is immersed for improved optical coupling into cell 25.
The optical trap is shown within cell 25 with particle 27 captured in the trap. Particle 27 is suspended in a liquid medium such as water, for example, which is enclosed by cell 25. Cell 25 is a transparent enclosure for enclosing the suspended biological particles or a transparent slide from which particle containing droplets can be hung. In one example, cell 25 has dimensions of 20 1 cm. x 3 cm. x 100 ,~m.
The position of cell 25 is adjustable in three dimensions (x, y, z) by the use of adjustable mount 2B. In practice, mount 26 is useful in locating and manipulating the biological particles.
Viewing of biological particles in the trap is accomplished directly or 25 through the use of a monitor. While other types of viewing such as viewing directly in cell 25 are possible, it is an added feature of the present invention the viewing is accomplished through the same lens objective which simultaneously creates the optical trap.
Illumination for viewing is provided by visible light source 2~ and is 30 projected through converging lens 28 onto the particles in the field of view.High resolution viewing occurs with the aid of lens 23 through which the visiblelight passes toward either the eyepiece or the monitor 18. For direct viewing, visible light shown as a dashed line is reflected from beam splitter 1~ to microscope eyepiece 21. Infrared blocking filter 22 is placed in front of eyepiece , ,, ... ~ ... .. .
:~3~ 7'S3 21 to isolate the viewing optics (viewer's eye) from back reflections from cell 25.
For monitoring, the visible light passes through beam splitter 1~ and is reflected from beam splitter 15 toward infrared blocking filter 17 and finally monitor 18.Infrared blocking rllter 17 isolates the monitor from back reflections from cell5 25.
In FIG. 2, the apparatus shown in FIG. 1 is augmented by a second infrared laser source and optics to create a second single-beam gradient force optical trap in cell 25. Infrared laser source 30 generates light beam 31 impinging on adjustably mounted diverging lens 32. Lens 32 causes beam 31 to 10 emerge in a diverging pattern as light beam 34. Adjustment of lens 32 is accomplished in three dimensions (x, y, z) via adjustable mount 33. Light beam 34 is reflected by mirror 35 which coincidently permits transmission of light beam 14. This would occur by judiciously choosing different wavelengths of operation for the separate laser sources. On the other hand, element 35 can be 15 realized as a beam splitter which would reflect approximately half of the light beam incident thereon and transmit the remaining half. As shown in FIG. 2, light beam 34 is converged by lens 23 to form a second trap in cell 25. Particle36 i9 confned in the second trap.
While not shown, it should now be apparent to those skilled in the art 20 that a second trap may be created in the cell by utilizing an additional set of optics including another high convergence microscope. The second trap may be created from light entering the cell on the side opposite the beam for the firsttrap or, for that matter, at any angle to the beam for the first trap.
Manipulation or orientation of particles is achieved by grabbing each end 2S of a rod-like particle, for example, and moving it at will.
In operation, it is necessary to move the trapped biological particles into the viewing plane. This is carried out by adjusting the position of the diverging lens or lenses. Similarly, translation, separation or isolation of the biological particles is easily affected by adjusting mount 26 by the desired amount.
FIGs. 3 through 5 show several modes of operation for the same optical trap. FIG. 3 shows the conventional mode of operation in which the focus of the beam from lens 23 lies within cell 25 and the trapping action relies on the backward gradient component of the optical force. Depending on the size of the particles, it is possible to trap up to approximately four or five particles within ~Z~
the trap at one time.
Both modes shown in FIGs. 4 and 5 require less intensity than for the trap in FIG. 3. In FIG. 4, the bottom plate of cell 25 provides the backward trapping force and the gradient provides the transverse trapping force. It is 5 possible to trap approximately twelve or more biological particles at one time.
In FIG. 5, the scattering force of the focused light beam provides transverse confinement due to its inward direction; backward trapping i9 supplied by the bottom plate of cell 25. In the latter mode of operation, it is possible to trapsignificantly greater numbers of particles than for the modes shown in FIGs. 3 10 and 4.
Various biological particles have been isolated, confined and transported in this type of optical trap. For example, some biological particles successful trapped are tobacco mosaic viruses (See Ashkin et al., Science, Vol. 235, pp.
1517-20 (1~87).), yeast, E. coli bacteria, blood cells containing hemoglobbin, and 15 complex cells or parts of cells containing chlorophyll structures.
In general, the biological particles investigate do not have the regular shape of the dielectric spheres studied earlier. For example, passive, string-like organisms were trapped wherein the organism was approximately 50 ,um long and approximately 1 ~m in diameter. In the case of tobacco mosaic virus, the 20 particles resemble a cylinder about 200 angstroms in diameter and 3100 angstroms long.
It is a significant attribute of the present invention that particle motility is preserved and reproductivity of the particles is maintained. Reproduction by trapped biological particles has been observed with offspring remaining in the 25 trap. In other words, the optical trap permits non-destructive manipulation of biological particles at optical powers approaching several hundred milliwatts.
It should be noted that the use of infrared light results in a lower intensity trap at the focal spot for the same laser power than for traps using visible light. However, the forces in the trap are approximately equal. Thus, 30 the infrared trap has the added benefit over visible light traps of inducing less local heating in the focal spot.
^
...... .. .
Claims (5)
1. Apparatus for generating a single-beam gradient force optical trap of particles, said apparatus comprising a laser for generating a light beam at a predetermined wavelength and means for focusing said light beam with sufficient convergence to form said optical trap in a predetermined region, said apparatus characterized in that said predetermined wavelength is substantially included in the infrared range ofwavelengths between 0.8 µm and 1.8 µm inclusively, so that said trap non-destructively confines at least one biological particle.
2. Apparatus as defined in claim 1 wherein said focusing means includes a lens having a numerical aperture greater than 0.9.
3. Apparatus as defined in claim 1 further including means for varying a position of said predetermined region.
4. Apparatus for generating a single-beam gradient force optical trap of particles, said apparatus being comprised of a laser for generating a light beam at a predetermined wavelength and means for focusing said light beam with sufficient convergence to form said optical trap in a predetermined region, said apparatus characterized in that said predetermined wavelength is substantially included in the infrared range ofwavelengths, so that said trap non-destructively confines at least one biological particle, said apparatus further including means for generating a second light beam substantially at the predetermined wavelength, said second light beam focused by said focusing means to form a second optical trap in a second predetermined region.
5. Apparatus as defined in claim 4 further including means for independently varying relative positions of the predetermined regions.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US07/098,120 US4893886A (en) | 1987-09-17 | 1987-09-17 | Non-destructive optical trap for biological particles and method of doing same |
| US098,120 | 1987-09-17 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1302753C true CA1302753C (en) | 1992-06-09 |
Family
ID=22267291
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000577697A Expired - Lifetime CA1302753C (en) | 1987-09-17 | 1988-09-16 | Non-destructive optical trap for biological particles and method of doing same |
Country Status (8)
| Country | Link |
|---|---|
| US (1) | US4893886A (en) |
| EP (1) | EP0307940B1 (en) |
| JP (1) | JPH0291545A (en) |
| AU (1) | AU585757B2 (en) |
| CA (1) | CA1302753C (en) |
| DE (1) | DE3852365T2 (en) |
| ES (1) | ES2064335T3 (en) |
| HK (1) | HK48996A (en) |
Families Citing this family (131)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5100627A (en) * | 1989-11-30 | 1992-03-31 | The Regents Of The University Of California | Chamber for the optical manipulation of microscopic particles |
| CA2031716C (en) * | 1989-12-07 | 1996-06-18 | Hiroaki Misawa | Laser microprocessing and the device therefor |
| US5198369A (en) * | 1990-04-25 | 1993-03-30 | Canon Kabushiki Kaisha | Sample measuring method using agglomeration reaction of microcarriers |
| US5079169A (en) * | 1990-05-22 | 1992-01-07 | The Regents Of The Stanford Leland Junior University | Method for optically manipulating polymer filaments |
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| US3710279A (en) * | 1969-12-15 | 1973-01-09 | Bell Telephone Labor Inc | Apparatuses for trapping and accelerating neutral particles |
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| US4327288A (en) * | 1980-09-29 | 1982-04-27 | Bell Telephone Laboratories, Incorporated | Method for focusing neutral atoms, molecules and ions |
-
1987
- 1987-09-17 US US07/098,120 patent/US4893886A/en not_active Expired - Lifetime
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1988
- 1988-09-08 JP JP63223675A patent/JPH0291545A/en active Granted
- 1988-09-14 AU AU22220/88A patent/AU585757B2/en not_active Ceased
- 1988-09-16 EP EP88115207A patent/EP0307940B1/en not_active Revoked
- 1988-09-16 CA CA000577697A patent/CA1302753C/en not_active Expired - Lifetime
- 1988-09-16 ES ES88115207T patent/ES2064335T3/en not_active Expired - Lifetime
- 1988-09-16 DE DE3852365T patent/DE3852365T2/en not_active Revoked
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1996
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| AU585757B2 (en) | 1989-06-22 |
| DE3852365T2 (en) | 1995-04-27 |
| JPH0291545A (en) | 1990-03-30 |
| DE3852365D1 (en) | 1995-01-19 |
| ES2064335T3 (en) | 1995-02-01 |
| US4893886A (en) | 1990-01-16 |
| EP0307940B1 (en) | 1994-12-07 |
| HK48996A (en) | 1996-03-29 |
| AU2222088A (en) | 1989-03-23 |
| JPH056136B2 (en) | 1993-01-25 |
| EP0307940A1 (en) | 1989-03-22 |
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