SYSTEMS FOR CHROMATIC ABERRATION CORRECTION IN TOTAL INTERNAL REFLECTION FLUORESCENCE MICROSCOPY

Correction elements that can be incorporated in objective-based TIRF microscopy instruments to correct for chromatic aberrations are described. A correction element can be placed between a multiple wavelength excitation beam source and the microscope objective lens. In one aspect, the thickness of the correction element is defined to compensate for different axial positions of the focal points associated with each excitation wavelengths traveling along the outer edge of lenses comprising a microscope objective lens. In another aspect, the correction element can be angled and/or configured so that the different wavelengths of multiple wavelength excitation light are shifted to adjust the angle of incidence for each wavelength at the specimen/substrate interface. 1. A fluorescent microscopy instrument to illuminate a specimen near a specimen/slide interface , the instrument comprising:an objective lens;a multiple wavelength light source to output a least two different wavelengths of excitation light directed to travel along a path located near edges of interior lenses of the objective lens; anda correction element to interact with the light transmitted between the source and the objective lens so that the light associated with each wavelength exits the objective lens to experience total internal reflected at a specimen/slide interface with a substantially uniform evanescent field penetration depth.2. The instrument of claim 1 , further comprising:an optical fiber with a first end and a second end, the first end coupled to the source to receive the at least two different wavelengths of light and output the light through the second end;a dichroic mirror to reflect the excitation into the path and transmit fluorescent light emitted from fluorescently labeled components of the specimen and collimated by the objective lens; anda detector to receive the fluorescent transmitted through the dichroic mirror.3. The instrument of claim 1 , wherein the correction element further ...

SYSTEMS FOR FLUORESCENCE ILLUMINATION USING SUPERIMPOSED POLARIZATION STATES

Various superimposing beam controls that can superimpose beams of light with different optical properties are described. In one aspect, a beam control receives a beam of light and outputs one or more beams. Each beam is output in a different polarization state and with different optical properties. Superimposing beam controls can be incorporated in fluorescence microscopy instruments to split a beam of excitation light into one or more beams of excitation light. Each beam of excitation light has a different polarization and is output with different optical properties so that each excitation beam can be used to execute a different microscopy technique. 1. A superimposing beam control comprising:a polarization rotating element to receive excitation light and to output the light in a desired polarization state;a first polarizing element to split the polarized light into a first beam with a first polarization state and a second beam with a second polarization state;two sets of focusing optics, each set of focusing optics to impart different optical properties to one of the beams; anda second polarizing element to superimpose the first and second beams to propagate in one direction.2. The beam control of claim 1 , wherein the polarization rotating element further comprises:a half-wave plate;a Fresnel Rhomb;a polarizer; anda motor to rotate the polarization rotating element to change the desired polarization of the light to be output from the polarization rotating element.3. The beam control of claim 1 , wherein the polarization rotating element is to select the polarization of the light to control the relative intensity of the first beam and the second beam.4. The beam control of claim 1 , wherein the first polarizing element and the second polarizing element are polarizing beamsplitters claim 1 , Wollaston prisms or Nomarski prisms.5. The beam control of claim 1 , wherein the first polarizing element is to split the polarized light so that the first polarization state ...

SYSTEM AND METHOD FOR CONTINUOUS, ASYNCHRONOUS AUTOFOCUS OF OPTICAL INSTRUMENTS

Embodiments of the present invention are directed to autofocus subsystems within optical instruments that continuously monitor the focus of the optical instruments and adjust distances within the optical instrument along the optical axis in order to maintain a precise and stable optical-instrument focus at a particular point or surface on, within, or near a sample. Certain embodiments of the present invention operate asynchronously with respect to operation of other components and subsystems of the optical instrument in which they are embedded. 123-. (canceled)24. An autofocus subsystem comprising:an autofocus light source;optical components that direct light from the autofocus light source into the optical path of an optical instrument;a focusing lens that focuses autofocus light returned to the autofocus system from the optical path of the optical instrument;{'sub': 'd', 'an autofocus detector arranged to detect the position Zof the focal point of the autofocus light along the optical-axis; and'}{'sub': 'd.', 'an autofocus processing component arranged to adjust the focus of the optical instrument in response to Z'}25. The autofocus subsystem of claim 24 , wherein the autofocus detector is arranged to perform a physical or logical optical-axis scan to detect Z26. The autofocus subsystem of claim 25 , wherein the autofocus detector comprises a fixed photodetector and an optical-axis scan member interposed between the focusing lens and the photodetector.27. The autofocus subsystem of claim 26 , wherein the optical-axis scan member is an autofocus rotor.28. The autofocus subsystem of claim 27 , wherein the autofocus rotor includes:a cylindrical rotor that is rotated about a central axis;a number of hollow, circularly disposed, cylindrical shafts that span the rotor in the direction of the central axis; and{'sub': 'd', 'an opaque disk with a central pinhole mounted within each cylindrical shaft at a particular distance from the top of the rotor corresponding to an ...

LIGHT-SCANNING SYSTEMS

Various light-scanning systems that can be used to perform rapid point-by-point illumination of a focal plane within a specimen are disclosed. The light-scanning systems can be incorporated in confocal microscopy instruments to create an excitation beam pivot axis that lies within an aperture at the back plate of an objective lens. The light-scanning systems receive a beam of excitation light from a light source and direct the excitation beam to pass through the pivot point in the aperture of the back plate of the objective lens while continuously scanning the focused excitation beam across a focal plane. 1. A light-scanning system comprising:a scanning mirror; andat least two stationary mirrors with reflective surfaces angled toward the scanning minor, wherein the scanning minor is positioned so that when a beam of light strikes the scanning minor, the beam is to undergo at least six reflections with a final reflection off of the scanning minor that places the beam on a path through a pivot point.2. The system of claim 1 , wherein the scanning minor is a flat minor to be rotated back and forth over a range of angles.3. The system of claim 1 , wherein the scanning mirror further comprises a galvanometer mirror.4. The system of claim 1 , wherein the scanning mirror further comprises a piezoelectric controlled minor.5. The system of claim 1 , wherein the at least two stationary minors comprise a first stationary minor with a reflective surface angled toward the scanning minor claim 1 , and a second stationary minor with a reflective surface angled toward the scanning minor and the first stationary minor.6. The system of claim 1 , wherein the at least two stationary minors further comprises two stationary minors arranged so that the beam is to undergo a first reflection off of the scanning mirror claim 1 , a second reflection off of a first mirror of the stationary mirrors claim 1 , a third reflection off of a second mirror of the stationary mirrors claim 1 , a fourth ...

LASER BEAM SELECTORS

Various beam selectors for selectively placing one of at least two beams of light along the same output path are disclosed. In one aspect, a beam selector receives at least two substantially parallel beams of light. The beam selector includes a plate with an aperture so that when one of the at least two beams is selected for transmission, the beam selector directs only the selected beam along an output path through the aperture. The plate can also serve to block transmission of unselected beams. The output path through the aperture is the same for each of the at least two beams when each beam is selected. Beam selectors can be incorporated into fluorescence microscopy instruments to selectively place particular excitation beams along the same path through the microscope objective lens and into a specimen to excite fluorescence of fluorescent probes attached to a particular component of the specimen. 1. A beam selector for selecting one of at least two beams of light comprising:a scanning mirror to receive the at least two beams of light;two stationary minors; anda plate with an aperture, wherein the scanning mirror is to be rotated into at least two positions, each position to select one of the beams to undergo a first reflection off of the scanning mirror, one reflection off of each of the stationary mirrors, and a second reflection off of the scanning mirror to place the beam on a path through the aperture.2. The selector of claim 1 , wherein the scanning mirror further comprises a flat mirror attached to a rotatable shaft of a motor and wherein the motor further comprises one of a galvanometer motor and a stepper motor.3. The selector of claim 1 , wherein the scanning mirror further comprises a piezoelectric controlled mirror.4. The selector of claim 1 , wherein the scanning mirror claim 1 , two stationary minors claim 1 , and plate are arranged so that a first of the stationary mirrors is angled toward a region between the scanning mirror and a second of the ...

LASER BEAM IRRADIANCE CONTROL SYSTEMS

Irradiance control systems (“ICSs”) that control the irradiance of a beam of light are disclosed. ICSs include in a beam translator and a beam launch. The beam translator translates the beam substantially perpendicular to the propagating direction of the beam with a desired displacement so that the beam launch can remove a portion of the translated beam and the beam can be output with a desired irradiance. The beam launch attenuates the irradiance of the beam based on the amount by which the beam is translated. ISCs can be incorporated into fluorescent microscopy instruments to provide high-speed, fine-tune control over the irradiance of excitation beams. 1. A system to control irradiance of a beam of light , the system comprising:a beam translator to translate the beam substantially perpendicular to a propagation direction of the beam with a desired displacement; anda beam launch located in the path of the translated beam, the launch to receive the translated beam and to attenuate the beam based on the amount by which the beam is to be translated.2. The system of further comprising:a feedback control electronically connected to the beam translator; anda splitter located in the path of the attenuated beam, the splitter to reflect a first portion of the attenuated beam to the feedback control and transmit a larger second portion, wherein the feedback control is to measure the irradiance of the first portion and direct the translator to translate the beam with the desired displacement based on the irradiance of the first portion.3. The system of claim 1 , wherein the beam translator includes:a scanning mirror; andat least two stationary mirrors, wherein the mirrors are positioned so that when the beam initially strikes the scanning mirror, the beam is reflected by the stationary mirrors and reflected a second time off of the scanning mirror to emerge on one of a continuum of substantially parallel paths.4. The system of claim 1 , wherein the beam undergoes four ...

SYSTEMS AND METHODS FOR ILLUMINATION PHASE CONTROL IN FLUORESCENCE MICROSCOPY

Illumination phase controls that provide precise and fast phase control of structured illumination patterns used in structure illumination microscopy are described. A coherent light source is used to generate a beam of coherent light that is split into at least three coherent beams of light. In one aspect, an illumination phase control is composed of at least one pair of rotatable windows to apply at least one phase shift to at least one of the beams. An objective lens is to receive the beams and focus the at least three beams to form an interference pattern. The phase control can be used to change the position of the interference pattern by changing the at least one phase shift applied to the at least one beam. 1. A microscopy instrument comprising:a splitter to receive a monochromatic beam of light and split the beam into at least three separate coherent beams;an illumination phase control including at least one rotatable window, the at least one rotatable window to apply at least one phase shift to the at least one beam; andan objective lens to focus the at least three beams to form an interference pattern, wherein the phase control to change position of the interference pattern with the at least one rotatable window to be rotated with respect to at least one path of the at least one beam.2. The instrument of claim 1 , further comprising an illumination pattern rotator disposed between the phase control and the objective lens claim 1 , the pattern rotator to rotate the at least three beams output from the phase control claim 1 , which results in the interference pattern to be rotated through the same angle.3. The instrument of claim 1 , wherein the splitter further comprises a one-dimensional grating.4. The instrument of claim 1 , wherein the splitter further comprises at least two beamsplitters.5. The instrument of claim 1 , wherein the illumination phase control including at least one rotatable window further comprises at least one pair of rotatable windows ...

Variable orientation illumination-pattern rotator

Variable orientation illumination-pattern rotators (“IPRs”) that can be incorporated into structured illumination microscopy instruments to rapidly rotate an interference pattern are disclosed. An IPR includes a rotation selector and at least one mirror cluster. The rotation selector directs beams of light into each one of the mirror clusters for a brief period of time. Each mirror cluster imparts a particular predetermined angle of rotation on the beams. As a result, the beams output from the IPR are rotated through each of the rotation angles imparted by each of the mirror clusters. The rotation selector enables the IPR to rotate the beams through each predetermined rotation angle on the order of 5 milliseconds or faster.

MICROSCOPY INSTRUMENTS WITH DETECTOR ARRAYS AND BEAM SPLITTING SYSTEM

Microscopy instruments with detectors located on one side of the instruments are disclosed. The microscopy instruments include a splitting system and an array of detectors disposed on one side of the instrument. A beam composed of two or more separate emission channels is separated by the splitting system into two or more beams that travel along separate paths so that each beam reaches a different detector in the array of detectors. Each beam is a different emission channel and the beams are substantially parallel. 2. The system of further comprising a polychroic mirror to reflect the channels to the splitting system.3. The system of claim 1 , wherein the splitting system includes:a set of mirrors; andone or more sets of optical filters, wherein each filter in each set is to reflect one of the channels toward one of the minors, and each minor is positioned and oriented to reflect the channel into one of the separate, substantially parallel paths.4. The system of claim 1 , wherein the set of the mirrors positioned around the set of filters further comprises the mirrors radially distributed around the set of optical filters.5. The system of claim 1 , wherein the optical filters are dichroic mirrors.6. The system of claim 1 , wherein the optical filters are polychroic mirrors.7. The system of claim 1 , wherein the splitting system is to direct the channels into separate claim 1 , substantially parallel paths further comprises the substantially claim 1 , parallel paths lie in the same plane.8. The system of claim 1 , wherein the splitting system is to direct the channels into separate claim 1 , substantially parallel paths further comprises the substantially claim 1 , parallel paths have a two-dimensional geometrical arrangement.9. The system of claim 1 , wherein the array of detector mounts is approximately planar.10. The system of claim 1 , wherein each detector further comprises one of a photodetector array claim 1 , a CCD camera claim 1 , or a CMOS camera.12. The ...

FLUORESCENCE MICROSCOPES WITH POLYCHROIC MIRROR CHANGERS

Fluorescence microscopy systems with polychroic mirror changers are described. In one aspect, a polychroic mirror changer includes a polychroic-mirror array. The array includes a plate with a planar surface and a number of holes formed in the plate. The array also includes polychroic mirrors attached to the planar surface of the plate such that each polychroic mirror covers one of the holes and the reflective surface of each polychroic mirror is adjacent to and aligned with the planar surface of the plate. Each polychroic mirror is partially exposed through a corresponding hole to reflect a different subset of excitation channels of a beam of excitation light input to the changer. The polychroic-mirror array can be mounted in the changer so that when a different subset of excitation channels is selected to illuminate a specimen, the plate is moved within a single plane of motion. 1. A microscopy system comprising:a light source to emit a number of excitation channels;a polychroic-mirror changer having a first opening to receive the excitation channels and a second opening; anda polychroic-mirror array positioned within the changer to intersect the excitation channels, wherein the array includes a planar arrangement of two or more polychroic mirrors, each polychroic mirror to reflect a different subset of the excitation channels through the second opening and into an objective lens to illuminate a specimen when the array is moved in a single plane of motion.2. The system of claim 1 , wherein the polychroic-mirror array includes a plate with a planar first surface and a second surface located opposite the first surface and has a number of holes that extend from the first surface to the second surface.3. The system of claim 1 , wherein each polychroic mirror is attached to the first surface and covers a hole such that each polychroic mirror is to receive the excitation channels through a corresponding hole and is to reflect a set of excitation channels back through the ...

IMPROVEMENTS IN AND RELATING TO CELL CULTURE MICROSCOPY SLIDES

Disclosed is a cell culture microscopy slide comprising an optically transparent generally flat supporting surface () including upper and lower opposed substrate surfaces (). A peripheral frame () surrounds the substrate (), the frame () having a lower frame surface () and an upper frame surface (). The lower frame surface () and the lower substrate surface () are generally flush. The upper frame surface () lies above the upper substrate surface (), to form a well (), and the upper and lower frame surfaces () are continuously flat and generally parallel. The substrate is preferably glass having a thickness of 1.7 mm. 1. A cell culture microscopy slide comprising:an optically transparent generally flat substrate including upper and lower opposed substrate surfaces; anda peripheral frame surrounding the substrate, the frame having a lower frame surface and an upper frame surface;wherein the lower frame surface and the lower substrate surface are generally flush;wherein the upper frame surface lies above the upper substrate surface, to form a well,wherein the upper and lower frame surfaces are generally parallel; andwherein the upper substrate surface includes areas coated with a hydrophobic material, and cell culture regions free from said material.2. A cell culture microscopy slide as claimed in wherein the slide further includes a machine readable slide identifier.3. A cell culture microscopy slide as claimed in wherein the slide further includes optical machine readable alignment indicia located in one corner of the frame.4. A cell culture microscopy slide as claimed in wherein the substrate is glass and has a generally uniform thickness of about 0.165 mm to about 0.175 mm claim 1 , and preferably about 0.170 mm thickness.5. A cell culture microscopy slide as claimed in wherein the upper and lower frame surfaces are continuously flat.6. A microscopy slide preparation method claim 1 , comprising the following steps:a) providing a cell culture microscopy slide ...

Mode-switchable illumination system for a microscope

Illumination system for a microscope system capable of being mode-switchable between a first and a second illumination mode, comprising one source of light for providing a collimated beam of light, at least one selector mirror capable of being positioned in at least two positions to redirect the beam of light in two different beam paths, the first beam path being a direct exit beam path wherein the selector mirror redirects the beam of light along an exit beam path to provide a first illumination mode, the second beam path is a mirror loop path comprising two or more mirrors arranged to redirect the beam of light onto the selector mirror such that it is redirected by the selector mirror a second time along the exit beam path, and wherein mirror loop path comprises at least one optical element arranged to optically alter the beam of light to provide the second illumination mode. According to one embodiment, the first illumination mode is Total Internal Reflection (TIRF) and the second illumination mode is Photokinetics (PK) illumination.

Improvements In and Relating to LED Illumination in Microscopy

Disclosed is an LED arrangement for a microscopy instrument ( FIG. ) comprising a light emitting area (), and a part-spherical solid and light transmissive cap (), in light communication with the light emitting area, the cap having a hemispherical surface () including a portion () at which light from the light emitting area is reflected and a portion () at which light from the emitter can exit the cap, in order to provide a usable light cone L which includes light recycled from the more divergent emitted light, and is thereby more intense. 1. An LED arrangement comprising a light emitting area , and a part-spherical , light transmissive cap , in light communication with the light emitting area , the cap having a curved surface including a portion at which light from the light emitting area is reflected and a portion at which light from the emitter can exit the cap.2. The LED arrangement as claimed in claim h wherein the curved surface of the cap is generally hemispherical and the cap further includes a generally flat surface across the diametric base of the hemispherical surface.3. The LED arrangement as claimed in claim 2 , wherein the light emitting area is proximal to a central region of the generally flat surface preferably held immediately adjacent thereto claim 2 , or in touching contact claim 2 , in each case to thereby provide said light communication and wherein said reflected light from the LED is reflected back toward the LED.4. The LED arrangement as claimed in claim 1 , wherein said reflecting portion of the curved surface includes a mirrored coating and said exit is not mirrored.5. The LED arrangement as claimed in claim 1 , wherein the light emitter is spaced from the exit by a distance such that the angle of divergence of light exiting the exit is less than 60 degrees claim 1 , preferably about 50 to 20 degrees claim 1 , and more preferably about 30 degrees.6. The LED arrangement as claimed in claim 1 , wherein the light emitter has a width w and the ...

SELECTIVE PLANE ILLUMINATION MICROSCOPY INSTRUMENTS

Disclosed is an optical arrangement providing selective plane illumination, including an inverted illumination objective mounted below a sample support in use providing a line or plane of light at the sample support, and at least one image collection objective mounted above the support, said inverted illumination objective having an illumination objective optic axis, and said image collection objective having an image collection objective optical axis, wherein illumination light is arranged to propagate toward the illumination objective lateral offset to the illumination objective optical axis such that the illumination light leaving the illumination objective propagates toward the sample support at an oblique angle relative to the illumination objective optical axis, and wherein the image objective optical axis has an angle α which is obtuse to the illumination objective optical axis and generally perpendicular to light propagating at the sample support. 1. An optical arrangement providing selective plane illumination , including an inverted illumination objective mounted below a sample support in use providing a line or plane of light at the sample support , and at least one image collection objective mounted above the support , said inverted illumination objective having an illumination objective optical axis , and said image collection objective having an image collection objective optical axis , wherein illumination light is arranged to propagate toward the illumination objective laterally offset to the illumination objective optical axis such that the illumination light leaving the illumination objective propagates toward the sample support at an oblique angle relative to the illumination objective optical axis , and wherein the image objective optical axis has an angle α which is obtuse to the illumination objective optical axis and generally perpendicular to light propagating at the sample support.2. An optical arrangement as claimed in claim 1 , wherein the ...

Polarization control systems

Polarization rotation controls that can be incorporated in structured illumination microscopy instruments to rotate an interference fringe pattern through a desired angle of rotation and rotate the polarization through substantially the same rotation angle are described. In one aspect, a polarization rotation control includes at least one polarization rotator and a splitter. A beam of polarized coherent light is transmitted through one of the at least one polarization rotator and the splitter. The splitter splits the beam into at least three coherent beams of light that are focus by an objective lens of the microscopy instrument to form an interference fringe pattern, and the splitter is rotated to rotate the interference pattern through a desired rotation angle. The at least one polarization rotator rotates the polarization of the light through substantially the same rotation angle.

Systems for fluorescence illumination using superimposed polarization states

Various superimposing beam controls that can superimpose beams of light with different optical properties are described. In one aspect, a beam control receives a beam of light and outputs one or more beams. Each beam is output in a different polarization state and with different optical properties. Superimposing beam controls can be incorporated in fluorescence microscopy instruments to split a beam of excitation light into one or more beams of excitation light. Each beam of excitation light has a different polarization and is output with different optical properties so that each excitation beam can be used to execute a different microscopy technique.

Light-scanning systems

Systems and methods for illumination phase control in fluorescence microscopy

Illumination phase controls that provide precise and fast phase control of structured illumination patterns used in structure illumination microscopy are described. A coherent light source is used to generate a beam of coherent light that is split into at least three coherent beams of light. In one aspect, an illumination phase control is composed of at least one pair of rotatable windows to apply at least one phase shift to at least one of the beams. An objective lens is to receive the beams and focus the at least three beams to form an interference pattern. The phase control can be used to change the position of the interference pattern by changing the at least one phase shift applied to the at least one beam.

Microscopy instruments with detector arrays and beam splitting system

Microscopy instruments with detectors located on one side of the instruments are disclosed. The microscopy instruments include a splitting system and an array of detectors disposed on one side of the instrument. A beam composed of two or more separate emission channels is separated by the splitting system into two or more beams that travel along separate paths so that each beam reaches a different detector in the array of detectors. Each beam is a different emission channel and the beams are substantially parallel.

Fluorescence microscopes with polychroic mirror changers

Fluorescence microscopy systems with polychroic mirror changers are described. In one aspect, a polychroic mirror changer includes a polychroic-mirror array. The array includes a plate with a planar surface and a number of holes formed in the plate. The array also includes polychroic mirrors attachedto the planar surface of the platesuch that each polychroic mirror covers one of the holes and the reflective surface of each polychroic mirror is adjacent to and aligned with the planar surface of the plate. Each polychroic mirror is partially exposed through a corresponding hole to reflect a different subsetof excitation channels of a beam of excitation light input to the changer. The polychroic-mirror array can be mounted in the changer so that when a different subsetof excitation channels is selected to illuminate a specimen, the plate is moved within a single plane of motion.

Fluorescence microscopes with polychroic mirror changers

Fluorescence microscopy systems with polychroic mirror changers are described. In one aspect, a polychroic mirror changer includes a polychroic-mirror array. The array includes a plate with a planar surface and a number of holes formed in the plate. The array also includes polychroic mirrors attachedto the planar surface of the platesuch that each polychroic mirror covers one of the holes and the reflective surface of each polychroic mirror is adjacent to and aligned with the planar surface of the plate. Each polychroic mirror is partially exposed through a corresponding hole to reflect a different subsetof excitation channels of a beam of excitation light input to the changer. The polychroic-mirror array can be mounted in the changer so that when a different subsetof excitation channels is selected to illuminate a specimen, the plate is moved within a single plane of motion.

System and method for continuous, asynchronous autofocus of optical instruments using multiple calibration curves

The current application is directed to autofocus subsystems within optical instruments that continuously monitor the focus of the optical instruments and adjust distances within the optical instrument along the optical axis in order to maintain a precise and stable optical-instrument focus at a particular point or surface on, within, or near a sample. Certain autofocus implementations operate asynchronously with respect to operation of other components and subsystems of the optical instrument in which they are embedded. The described autofocus subsystems employ multiple calibration curves to precisely adjust the z-position of an optical instrument.

System and method for continuous, asynchronous autofocus of optical instruments using multiple calibration curves

The current application is directed to autofocus subsystems within optical instruments that continuously monitor the focus of the optical instruments and adjust distances within the optical instrument along the optical axis in order to maintain a precise and stable optical-instrument focus at a particular point or surface on, within, or near a sample. Certain autofocus implementations operate asynchronously with respect to operation of other components and subsystems of the optical instrument in which they are embedded. The described autofocus subsystems employ multiple calibration curves to precisely adjust the z-position of an optical instrument.

Mode-switchable illumination system for a microscope

Illumination system for a microscope system capable of being mode-switchable between a first and a second illumination mode, comprising one source of light for providing a collimated beam of light, at least one selector mirrorcapable of being positioned in at least two positions to redirect the beam of light in two different beam paths, the first beam path being a direct exit beam path wherein the selector mirror redirects the beam of light along an exit beam path to provide afirst illumination mode, the second beam path is a mirror loop path comprising two or more mirrors arranged to redirect the beam of light onto the selector mirror such that it is redirected by the selector mirror a second time along the exit beam path, and wherein mirror loop path comprises at least one optical element arranged to optically alter the beam of light to provide the second illumination mode. According to one embodiment,the first illumination mode is Total Internal Reflection (TIRF)and the second illumination mode is Photokinetics(PK) illumination.

System and method for continuous, asynchronous autofocus of optical instruments

Embodiments of the present invention are directed to autofocus subsystems within optical instruments that continuously monitor the focus of the optical instruments and adjust distances within the optical instrument along the optical axis in order to maintain a precise and stable optical-instrument focus at a particular point or surface on, within, or near a sample. Certain embodiments of the present invention operate asynchronously with respect to operation of other components and subsystems of the optical instrument in which they are embedded.

Selective plane illumination microscopy instruments

Disclosed is an optical arrangement providing selective plane illumination, including an inverted illumination objective mounted below a sample support in use providing a line or plane of light at the sample support, and at least one image collection objective mounted above the support, said inverted illumination objective having an illumination objective optic axis, and said image collection objective having an image collection objective optical axis, wherein illumination light is arranged to propagate toward the illumination objective lateral offset to the illumination objective optical axis such that the illumination light leaving the illumination objective propagates toward the sample support at an oblique angle relative to the illumination objective optical axis, and wherein the image objective optical axis has an angle α which is obtuse to the illumination objective optical axis and generally perpendicular to light propagating at the sample support.

Laser beam selectors

Various beam selectors for selectively placing one of at least two beams of light along the same output path are disclosed. In one aspect, a beam selector receives at least two substantially parallel beams of light. The beam selector includes a plate with an aperture so that when one of the at least two beams is selected for transmission, the beam selector directs only the selected beam along an output path through the aperture. The plate can also serve to block transmission of unselected beams. The output path through the aperture is the same for each of the at least two beams when each beam is selected. Beam selectors can be incorporated into fluorescence microscopy instruments to selectively place particular excitation beams along the same path through the microscope objective lens and into a specimen to excite fluorescence of fluorescent probes attached to a particular component of the specimen.

Variable orientation illumination-pattern rotator

Variable orientation illumination-pattern rotators (“IPRs”) that can be incorporated into structured illumination microscopy instruments to rapidly rotate an interference pattern are disclosed. An IPR includes a rotation selector and at least one mirror cluster. The rotation selector directs beams of light into each one of the mirror clusters for a brief period of time. Each mirror cluster imparts a particular predetermined angle of rotation on the beams. As a result, the beams output from the IPR are rotated through each of the rotation angles imparted by each of the mirror clusters. The rotation selector enables the IPR to rotate the beams through each predetermined rotation angle on the order of 5 milliseconds or faster.

Microscopy instruments with beam splitting system including optical filters and mirrors

Microscopy instruments with detectors located on one side of the instruments are disclosed. The microscopy instruments include a splitting system and an array of detectors disposed on one side of the instrument. A beam composed of two or more separate emission channels is separated by the splitting system into two or more beams that travel along separate paths so that each beam reaches a different detector in the array of detectors. Each beam is a different emission channel and the beams are substantially parallel.

Light-scanning systems

Various light-scanning systems that can be used to perform rapid point-by-point illumination of a focal plane within a specimen are disclosed. The light-scanning systems can be incorporated in confocal microscopy instruments to create an excitation beam pivot axis that lies within an aperture at the back plate of an objective lens. The light-scanning systems receive a beam of excitation light from a light source and direct the excitation beam to pass through the pivot point in the aperture of the back plate of the objective lens while continuously scanning the focused excitation beam across a focal plane.

Laser beam selectors

Various beam selectors for selectively placing one of at least two beams of light along the same output path are disclosed. In one aspect, a beam selector receives at least two substantially parallel beams of light. The beam selector includes a plate with an aperture so that when one of the at least two beams is selected for transmission, the beam selector directs only the selected beam along an output path through the aperture. The plate can also serve to block transmission of unselected beams. The output path through the aperture is the same for each of the at least two beams when each beam is selected. Beam selectors can be incorporated into fluorescence microscopy instruments to selectively place particular excitation beams along the same path through the microscope objective lens and into a specimen to excite fluorescence of fluorescent probes attached to a particular component of the specimen.

Fluorescence microscopes with polychroic mirror changers

Fluorescence microscopy systems with polychroic mirror changers are described. In one aspect, a polychroic mirror changer includes a polychroic-mirror array. The array includes a plate with a planar surface and a number of holes formed in the plate. The array also includes polychroic mirrors attached to the planar surface of the plate such that each polychroic mirror covers one of the holes and the reflective surface of each polychroic mirror is adjacent to and aligned with the planar surface of the plate. Each polychroic mirror is partially exposed through a corresponding hole to reflect a different subset of excitation channels of a beam of excitation light input to the changer. The polychroic-mirror array can be mounted in the changer so that when a different subset of excitation channels is selected to illuminate a specimen, the plate is moved within a single plane of motion.

Mode-switchable illumination system for a microscope

Illumination system for a microscope system capable of being mode-switchable between a first and a second illumination mode, comprising one source of light for providing a collimated beam of light, at least one selector mirror capable of being positioned in at least two positions to redirect the beam of light in two different beam paths, the first beam path being a direct exit beam path wherein the selector mirror redirects the beam of light along an exit beam path to provide a first illumination mode, the second beam path is a mirror loop path comprising two or more mirrors arranged to redirect the beam of light onto the selector mirror such that it is redirected by the selector mirror a second time along the exit beam path, and wherein mirror loop path comprises at least one optical element arranged to optically alter the beam of light to provide the second illumination mode. According to one embodiment, the first illumination mode is Total Internal Reflection (TIRF) and the second illumination mode is Photokinetics (PK) illumination.

Systems and methods for illumination phase control in fluorescence microscopy

Illumination phase controls that provide precise and fast phase control of structured illumination patterns used in structure illumination microscopy are described. A coherent light source is used to generate a beam of coherent light that is split into at least three coherent beams of light. In one aspect, an illumination phase control is composed of at least one pair of rotatable windows to apply at least one phase shift to at least one of the beams. An objective lens is to receive the beams and focus the at least three beams to form an interference pattern. The phase control can be used to change the position of the interference pattern by changing the at least one phase shift applied to the at least one beam.