BRPI0413573B1 - OPTICAL DETECTION SYSTEM FOR OBSERVING A MICROFLUID SYSTEM FOR FORMATING A SINGLE INCIDENT BEAM, FOR INTERROGATING A MICROFLUID SYSTEM AND OPTICAL SYSTEM - Google Patents

Description

"OPTICAL DETECTION SYSTEM FOR OBSERVING A MICROFLUIDIC SYSTEM FOR FORMING A SINGLE INCIDENT BEAM FOR INTERROGATING A MICROFLUIDIC SYSTEM AND OPTICAL SYSTEM" Related Applications August 2003 and US Patent Application No. XX / XXX.XXX, filed August 9, 2004, the contents of which are expressly incorporated in their entirety by that reference.

Field of the Invention The present invention relates to a system and method for monitoring particles that flow through a channel.

Background of the Invention In a system, such as a micro fluid system, that carries particles through one or more channels, an optical system can be used for particle monitoring, analysis or detection. Optical systems may be useful, for example, in particle separation systems, which separate a particle stream that flows through one or more channels based on a predetermined characteristic.

Conventional detection systems have significant disadvantages. For example, earlier optical detection systems are sometimes inaccurate and provide poor results due to the difficulty of observing low light level signals from particulate fluorescent labels when spread over a large area. Earlier optical systems also have difficulty when the light signals to be detected are short lived, for example less than one millisecond. For example, conventional charge-coupled device (CCD) technology has a frame rate of more than one millisecond.

Earlier micro channel interrogation systems are also limited to focusing light on a single channel, a region of less than about 500 µm, and capturing light from a limited region in a similar manner.

SUMMARY OF THE INVENTION The present invention provides an optical system for the acquisition of fast spatially shaped channel array spectra. The system is designed to be used to interrogate a micro fluidic particle analysis or separation chip containing a set of one or more parallel fluidic channels spaced 1 to 200 millimeters apart. Particles carried in the channels have speeds of 0.1 to 10 meters per second, so the signals observed by detectors may have a duration of less than one millisecond and may require observation with detectors and 1 to 100 bandwidth electronics. megahertz. The optical detection system includes a light source for producing a light beam passing through the micro fluidic chip or the channel can be monitored, one or more lenses or optical fibers to capture light from the light source after interaction. with particles or chemicals in the micro fluid channels, and one or more detectors. Detectors, which may include light amplifying elements, detect each light signal and transform the light signal into an electronic signal. Electronic signals, each representing the strength of an optical signal, pass from each detector to an electronic data acquisition system for analysis. The light amplifying element or elements may comprise a phototube array, a multiple anode phototube or an image intensifier-based multi-channel plate coupled to a photodiode detector array. The optical system inexpensively and simultaneously captures extinction signals, one or more scattered optical signals, one or more fluorescent signals, all at low light and with high bandwidth (> 1 MHz) from a set of one or more. Particle transport channels at one time. The system provides efficient and accurate monitoring of each particle under various conditions.

BRIEF DESCRIPTION OF THE FIGURES The invention will be apparent from the description and accompanying drawings, in which similar reference characters refer to the same parts throughout the different views. Figure 1 illustrates a system having a plurality of channels for carrying particle streams suitable for implementing an illustrative embodiment of the present invention; Figure 2 is a schematic diagram of an optical detection system of the present invention; Figure 3 illustrates a cross section through a micro channel in a plane perpendicular to the micro channel; Figure 4 is a schematic diagram of an optical detection system of the present invention illustrating in detail the components of the fluorescence detector; Figure 5 illustrates an optical detection system suitable for particle analysis on a plurality of channels of a micro fluidic system;

Figures 6a to 6c illustrate a subsystem embodiment for optical detection scattered at a 90 degree angle or extinction in the optical detection system of Figure 2; Figure 7 is a beamforming optical scheme suitable for use in the optical detection system of Figure 2; Figure 8 illustrates a segmented mirror suitable for use in the optical detection system of the present invention; Figure 9 is a partial view of a groove of the segmented mirror of Figure 8; Figure 10 is a table illustrating different configurations for a segmented mirror groove based on a corresponding local width; Figure 11 is a schematic of beam format optics employing a segmented mirror in an optical detection system of an illustrative embodiment of the invention; Figure 12 illustrates an image intensifier suitable for use with the optical detection system of an illustrative embodiment of the invention.

Detailed Description of the Invention The present invention provides an optical system for monitoring and detecting particle flow through a set of channels. The present invention will be described below with respect to illustrative embodiments. Those skilled in the art will appreciate that the present invention may be implemented in a number of different applications and embodiments and is not specifically limited in its application to the particular embodiments set forth herein. Figure 1 illustrates a micro fluid system 10 suitable for implementing an illustrative embodiment of the invention including a plurality of channels for carrying a substance such as particles or cells. Illustrative micro fluidic system 10 comprises a substrate 1 having a plurality of channels, such as micro channels 3, disposed therein. The channels carry fluid and / or particles through the micro fluid system 10 to process, handle and / or perform any suitable operation on a liquid sample. As used herein, the term "micro fluidic" refers to a system or device for the handling, processing, ejection and / or analysis of a fluid sample including at least one channel having micro scale dimensions. The term "channel" as used herein refers to a pathway formed in or through a medium that allows movement of fluids such as liquids and gases. The term "micro channel" refers to a channel preferably formed in a micro fluidic system or device having transverse dimensions in the range from about 1.0 pm to about 500 pm, preferably from about 25 pm to about 350 pm and more preferably between about 50 pm and about 300 pm. Those skilled in the art will be able to determine an appropriate channel volume and length. The ranges should include the values mentioned above as upper and lower limits. The channel may have any selected shape or arrangement, examples of which include a linear or nonlinear configuration and a U-shaped configuration. The micro fluid system 10 may comprise any suitable number of micro channels 3 for transporting fluids through the system. The present invention provides an optical detector for use with a micro fluid chip, such as the micro fluid system of Figure 1. The optical detector of the present invention may be implemented in a measuring region 2 of the micro fluid system for interrogation. the system in that region. The invention provides the ease of construction of a detection system that can scale micro-fluid chips with parallel channel sets of 1 to 200 channels arranged over one or more interrogation regions 2, which have a physical extension of 1 to 250 mm in length. from 1 to 100 mm. The optical detector can monitor flow through a plurality of channels on the chip simultaneously. The optical detector or optical detector system may inspect individual particles for one or more particular characteristics, such as size, shape, fluorescent intensity optical scattering, and other features obvious to those skilled in the art. For example, in one illustrative embodiment, the optical detector of the present invention may be positioned over a relatively large area of the chip (e.g., an active area of about 12mm to 50mm in diameter) containing more than one hundred particle channels. in flow to be observed. The optical detector is capable of capturing fast, low light signals from a plurality or all channels at once. Those skilled in the art will recognize that the optical system is not limited to use in particle or cell separation systems and may be implemented in any suitable system having a substance, such as particles, to be monitored by flowing through one or more channels. Figure 2 illustrates an overview of an optical detection system 8 of an illustrative embodiment of the invention which may be implemented in the micro fluidic system of Figure 1. Those skilled in the art will recognize that the optical detection system may be implemented in any system. available and is not limited to the micro fluidic system of Figure 1. Optical sensing system 8 includes a light source 11, illustrated as a laser, coupled with beamforming optics 12 to produce and form a passing light 14 through an optical mask 13, illustrated as a set of holes aligned with a set of particle transport channels 3 on the micro fluidic chip 10. The light admitted through the holes subsequently passes through the transport channels 3 themselves. The beam of light admitted to each channel through one or more associated holes forms an intersection with the particles 18 which are transported through channel 3 to create the optical signals. Examples of optical signals that may be produced in the analysis, cytometry or separation of optical particles when a light beam intersects with a particle include optical extinction, angle dependent optical scattering and fluorescent light. Optical extinction refers to the amount of light that passes through the particle without interaction. Angle-dependent optical scattering refers to the fraction of light that is scattered or bent at each angle (theta) away from the incident light beam. Fluorescent light is light that is absorbed by the molecules in the particle and emitted again with a longer wavelength. The detector optics 15, 15, 17, located on an opposite side of channel 3 of light source 11, captures and observes the optical signals generated by the intersection of a light beam with a particle in a channel. Optical Extinction detectors 15 are located directly opposite light source 11 and aligned with incident light path 14 for optical extinction detection. Optical scatter detectors 16 are located substantially perpendicular to the incident light path 14 in the plane formed by the incident light vector and the intersecting micro fluidic channel. Preferably, the optical scatter detectors are located at an angle of about 90 degrees to the incident light path 14. Optical Scatter detectors for other angles may optionally be located at these angles in the same plane. A fluorescence detection subsystem 17 captures optical fluorescence signals. The fluorescence detection subsystem 17 may include a large high numerical aperture lens and attached optical elements. As illustrated, the fluorescence detection subsystem is located above the micro-fluidic chip 10 to capture as many fluorescent photons as possible and reproduce its image on detectors (not shown). Optical detection system 8 may be implemented in an interrogation area 2 of chip 10. Illustrative interrogation area 2 encompasses 24 channels 3, although those skilled in the art recognize that any suitable number of channels can be observed using the optical detection system 8. In the illustrative embodiment, the interrogation area 2 is about 10 mm wide (across a plurality of channels 3) by 4 mm long (along each channel 3), although those versed in the recognize that the invention is not limited to this range.

When light 14 from a laser 11 or other optical source shines on chip 10, only light passing through the narrow region that the particles follow can interact with the particles to produce an optical signal. Light passing through chip 10 out of channels 3 or light passing through a region of a channel that contains no particles can only contribute to the background or noise and not to the signal and therefore is lost light and should be minimized. It is also a consideration that light passing through the chip without passing through the particles to be observed represents wasted laser source energy and should therefore be minimized for cost and temperature management reasons. Optical mask 13 formed by the hole layer and beam-forming optics 12 minimize lost light and minimize laser energy waste.

As illustrated, light source 11 provides incident light at about a 45 degree angle to channel 3. Thus, forward spread / extinction extends in the same direction on the opposite side of channel 3. As illustrated, the lead scatter 14b extends at an angle of 45 degrees from channel 3. Side scatter 14c extends about 90 degrees from incident light, providing fluorescence optics 17 with a mechanical freedom cone 170. The freedom cone Mechanical 170 provides a 90 degree unobstructed view to the detector between feed spread 14b and side spread 14c. Figure 3 illustrates an illustrative cross-sectional image through a portion of a micro fluidic chip 10 containing a pair of micro channels 3a and 3b. The cross section is in a plane that cuts through the micro channels and the holes 13a, 13b of the mask 13. The incident light 14 is partially blocked by the hole layer 13 and narrows the initial beam 14 to the focused beams 18 defined by each hole. 13a, 13b. The focused beams 18 form an intersection with each channel to illuminate the region 31 in which the particles 18 may flow in a conventional core stream. Much of the lost light is blocked by the hole layer 13, which may be a separate part of the micro-fluid chip or may be fabricated on the chip surface by photolithography or other methods known to those skilled in the art of chip manufacturing. The micro-fluidic system may comprise any system including channels for flowing a substance, such as particles or cells, therethrough. For example, the micro fluidic system 10 may comprise a particle separation system, such as the particle separation systems described in U.S. Patent Nos. 10 / 179,488 and 10 / 329,008, the contents of both patent applications are incorporated herein by reference. Other suitable micro fluidic systems are described in U.S. Patent Applications Nos. 10 / 028,852, 10 / 027,484, 10 / 027,516 and 10 / 607,287, all of which are incorporated herein by reference. Figure 4 illustrates a schematic diagram of the optical detection system of Figure 2 illustrating in detail the components of fluorescence detection subsystem 17. Fluorescence detection subsystem 17 includes a configured high (low F #) numeric aperture collection lens 45 and positioned to capture as many photons emitted from the illuminated particle as possible. Lens 45 may be a commercially available focal length 50mm F # = l ready lens. An example is the Leica Nocti-lux 50mm F # l lens. Larger lenses are also available and in use for multi-slot plate imaging. A scatter element 46, illustrated as a Lit-trow prism, is located above the first collection lens 45. The scatter element 46 bends light in a manner related to the wavelength of the particular beam. Illustrative Littrow prism 46 is 6.2 mm in diameter with an active area of 73 mm. The Littrow 46 prism has 720 grooves / mm and has a brightness angle of 43.1 degrees at 550 nm (the angle at which the prism is opposite the vertical plane). The Littrow angle is 23.33 degrees which is the angle at which the 550 nm light is bent away from the vertical plane in Figure 4. Those skilled in the art will recognize that any suitable means for bending the light in a particular way. may be used in accordance with the teachings of the invention. A reconstruction lens 47 is positioned at the Littrow angle to capture 1st order diffraction light from prism 46 and reconstruct the diffracted light into an illuminated particle image in the image plane 48.

A fiber assembly 49 extends from image plane 48 and carries signals to detectors 50 for signal analysis. The detectors may be a camera or other suitable device.

Due to the presence of the Littrow prism in the optical path, the illuminated particle in micro channel 3 is represented in plane 48 with longer wavelength photons inclined at an angle greater than shorter wavelength photons so that the particle have a spread spectrum across that image plane. Photons having a wavelength of from 500 nm to 700 nm are scattered across about 7841 microns in the image plane 48 for the 50 mm focal length lenses used for the slow 45 and 47. The illustrative embodiment has a spectral resolution 39.2 microns per nm wavelength. Optical detection system 8 may be used to observe antibody-labeled particles attached to fluorophores or other particle fluorescent markers known to those skilled in the cytometric art. When the excitation light is 488 nm in wavelength, then, for example, antibody labeled particles bound to the fluorophores FITC (fluorescein isothiocyanate), PE (R-phycoerythrin), APC (AlloFicoCyanin) and PerCP ( Peridinin-chlorophyll-protein complex) which exhibit peak fluorescent emission at 530 nm, 575 nm, 630 nm, and 695 nm, respectively. The FITC, PE, and PerCP photons are located in the image plane at positions -784 microns, 980 microns, 3136 nm, and 5684 microns (with respect to 0 to 550 nm), respectively. An opaque plate with 400 µm holes and 400 µm diameter optical fibers located therein will then provide each fiber 49 with a wavelength capture bandwidth of about 10 nm. Placing a fiber 49 at each location corresponding to the desired peak fluorophor emission produces an efficient and compact multi-color detection system. The fibers 49 located at one end in the image plane 48 have their other end attached to a detector. In the illustrative embodiment, the second end of the fibers is coupled to the photocathode window of a phototube (e.g., a single anode H6780-20 or Hamamatsu Inc. 32-anode phototubes) at a location corresponding to a single anode, in order to amplify fluorescent optical signals and convert them to electronic signals. Other amplifier light detectors such as image intensifiers or avalanche photodiode assemblies or others known to those skilled in the optical art may also be used to detect optical signals and convert them to electronic signals.

In Figure 4, the fibers 49 interrogating the particles in the illustrated channel are located in the same plane as the channel plane in the micro fluidic chip. If the system is used in a multi-channel set then the other channels are in front of the illustrated channel plane or behind the illustrated channel plane. Figure 5 illustrates a perspective view of an optical detector system 80 used to observe multiple channels on a micro fluidic chip. The optical detector system 80 also includes a set of holes 13 blocking most incident light 14 and illuminating small detection regions 2 in each channel 3 of the six channels of the micro fluidic chip. The collection lens optics, Littrow prism, and reconstruction lens are similar to those shown in Figure 4, and may have the same modalities as the lens and mesh specifications. In general, the size of lens components and network assemblies should be sufficient to provide a field of view on the chip that exceeds the size of the detection region (the region where channels are illuminated through the holes). In image plane 48 a board 480 is located holding six sets 490, including four fibers each. Each set of four optical fibers 49 is positioned to sample the optical spectrum emitted from an associated channel 3. Each fiber in the set is positioned at the peak emission site of a fluorophor. High numerical aperture fibers or lens fibers are suitable herein as will be apparent to those skilled in the art.

Figures 6a to 6c illustrate a subsystem embodiment for detecting optical scattering at a 90 degree angle or extinction. In this embodiment, an optical extinction column detector tape 63 is positioned above a multi-channel chip 10 with an intercanal spacing of about 500 microns. Optical extinction detector detector tape 63, a cross-section of which is illustrated in Figure 6b, is a mechanical part with 300 micron diameter holes drilled thereto at a depth of less than the thickness of tape 63d, and spaced by 500 microns in the centers to align the holes with the channel spacing. A high numerical aperture fiber 65 is placed in each hole to form a fiber assembly 61 with one fiber per channel. A hole with smaller diameter but concentric columns with the 63c fiber hole is drilled in each hole. This column hole penetrates the ribbon connector 63b, and allows light to pass through the column hole 63c and into the fiber 65 positioned on the larger diameter shaft. To make this system work, incident light 68 intersects the hole and channel at an angle of nearly 45 degrees and optical extinction detection tape 63 is mounted directly along the incident light vector (this is at an angle of 180 degrees to the incident light) as illustrated by the position of the tape. The aperture of the column former should exceed the aperture of the hole so that for a well-focused incident light all light crossing the hole can be detected in the fiber at the end of the column former. The speaker former itself is chosen to be long enough to reject any lost light from other channels. For example, in one embodiment, the hole aperture is 150 microns in diameter, the column former is 250 microns in diameter, the fiber is 300 microns in diameter, and the collimator, which is positioned within 2 mm of the channel, has 1 mm. of lenght. At the furthest end of the fiber assembly 61, each fiber is attached to a light tube or other optical detector. Optical extinction is often strong enough to use a photodiode for its detector.

In Figure 6c, a second tape 66 constructed substantially the same as the first described tape 63 but positioned at 90 degrees from incident light that is suitable for measuring 90 degree scattering or lateral spacing signals from cells or particles. Those skilled in the art will recognize that similar ribbons may be positioned at other angles to observe scattering parameters. A particular angle of interest is the so-called advance scattering which is the optical scattering in the almost straight direction generally closest to the forward feed positioning (almost 180 degrees of incidence) without direct light acquisition in the extinction path.

In an additional embodiment, the light source 11 is a Coherent Sapphire 488/200 laser which is a small, air-cooled solid state device producing about 200 nw with little or no noise from gas laser tube emissions. Alternatively, an optically pumped solid state (OPSS) laser is used which is also capable of generating all the different different excitation wavelengths required for monitoring. Those skilled in the art will recognize that any suitable light source may be used. Figure 7 is a cross-sectional view of a beamforming optic embodiment 12 suitable for use with the optical detector of the illustrative embodiment of the invention. The optical scheme is drawn on the x-z plane with the general direction of light propagation along the z axis. Each dotted line leads to a light beam x-y profile representation 14 'to illustrate how the beam is manipulated by the formatting optics. The beam passes from a single laser output 11 of an almost round profile of 700 microns in diameter to a wavelength filtered beam after a low pass filter or bandpass 74. The beam then passes through a first pair. of cylindrical collimation lenses 73 having a focal length of 5 mm having a focal length of 250 mm which produces a substantially rectangular shaped beam. The beam then passes through a focus lens 71 having focal length which is a 150 mm cylindrical lens to sharpen the beam 14 to 100 microns on the y axis. The overall profile in this embodiment after focus lens 71 is 36 mm by 100 microns and can be used to illuminate a pin set 13 of up to seventy holes / channels in the 500 micron spacing. Since the holes are less than about 100 microns in the y-axis direction, beam limitation prevents wastage of light. On an N-hole chip spaced by 500 microns in the centers it is preferable that the beam is slightly larger than 500 XN microns along the x axis and 200 microns along the y axis (slightly larger than 100 microns) in order to minimize the wasted laser energy. The columnar and formatted beam then intersects with the set of holes 13 and becomes N-shaped formatted beam 78 which are spaced to form an intersection with the coincident set of channels 3. The beam formatting mode of Figure 7 is very usable allowing for a minimum of lost light and acceptable energy efficiency of about 10% whereas this design allows simultaneous observation of rapid extinction (> 10 MHz bandwidth), scattering and fluorescence of many channels at once. Figure 8 illustrates a reflective beam splitter 80 based on a grooved mirror suitable for use in the optical detection system of the present invention. The beam splitter 80 includes a segmented mirror 83 for splitting an incoming light beam into a plurality of beams. An incident beam with columns 82 enters divider 80 and is reflected off an incident mirror 81 which is used to set the correct incident angle (usually a low angle) to the beam in segmented mirror 83 which divides the incident mirror. incident beam in a smaller beam array 84. The smaller beam array 84 extends upward in parallel with the incident beam 82. The segmented mirror 83 comprises a uniform set of reflective grooves. Preferably, the uniform assembly comprises anisotropically etched silicon. Alternatively, the uniform groove assembly is made of conventionally machined metal with an optical finish. In another embodiment, the uniform set of grooves is formed of a plastic material which is then covered with a reflective coating for the set of grooves. Figure 9 illustrates the angles and formulas guiding the design of such segmented mirrors. The incident beam 82 is partially secured by each groove 83a in the spread and that trapped portion is reflected outwardly at a fixed angle to create a narrower beam 84a. A second narrow beam 84b is formed by an adjacent groove 84b. Each groove is separated by groove spacing A and the divider generates beams of uniform local width (assuming uniform grooves) and the beam or spacing L that is designed to match the hole and channel spacing on the micro fluidic chip. Figure 10 is a table of modalities of the beam splitter of Figure 8 where the row spacing L is 500 microns and the grooves are fabricated with anisotropic silicon etching (which has a fixed groove angle e = 54.74). The table indicates a mirror setting suitable for a selected local size. For example, a local size of 100 microns is suitable for a spindle of <100 microns, and corresponds to a groove spacing A = 575 microns, groove slope G = 2,9, 7 degrees and an incidence angle of I = 25 degrees . Figure 11 depicts one embodiment of beam format subsystem 112 suitable for use in the optical detector system. Illustrative beam formatting subsystem 112 makes use of a segmented mirror 80, such as the segmented mirror of Figure 8, at a final stage after employing similar beam format optics 12 similar to beam format optics 12 described with relation to Figure 7.

An alternative embodiment includes fabricating hole assemblies 13 on each micro fluidic chip rather than mounting them separately on the optical system.

An alternative embodiment of the fiber set detectors used in the image plane of Figures 4 and 5 is to place an image intensifier in that plane and place the fibers behind that image intensifier to read the optical signal it produces in its fluorescent substance. Such an alternative can reduce costs by using only one light amplifier element (the image intensifier) for all fluorescence signals, and then photodiected for converting the after image intensifier optical signals into electronic signals. Figure 12 illustrates an image of a standard Hamamatsu image intensifier 220 but those skilled in the art will recognize that any large area light amplification component with high spatial resolution can be used in this alternative. Image intensifier 220 is used to amplify the intensity of an optical image before passing the signal to a photodiode array or other appropriate detection device. As illustrated, the image intensifier includes an input window 221 for the image signal, a light sensitive electron emitter, such as a photocathode 222, for transforming light into photoelectrons, an MCP 223 for electron multiplication, a screen of fluorescent substance 224 to convert the electrics into light and an exit window 225, illustrated as a fiber optic plate. According to an illustrative embodiment, the image intensifier may comprise a 25mm to 40mm Hamamatsu image intensifier, although those skilled in the art recognize that any suitable device may be used.

An alternative embodiment for both beam formatting subsystem 12 and fluorescence detection subsystem 17 includes short pass or long pass filters or wavelength band pass or band lock to remove stray source light or spurious in case fluorescence detection system or to remove lost or spurious wavelength components from light emitted by the light source 11.

An alternative embodiment of extinction and scatter detectors 15 and 16 is to add an independent laser energy monitor to the system for use in normalizing these signals. This is useful as both signals are directly proportional to laser energy so that laser noise can distort these signals.

An alternative embodiment to the fiber assemblies used with detectors 15, 16 and 17 is the replacement of each fiber array with an avalanche photodiode or photodiode array or other optical detector assembly. Those skilled in the art will recognize that alternative detectors are possible here as long as they match the light level requirement of the samples and form the factor requirements of the specific chip modalities to be used.

An alternative embodiment of the beam splitter may utilize reflective groove assemblies manufactured by anisotropic etching of crystalline materials or conventional metal machining or plastic forming followed by proper optical polishing or reflective coating.

In all embodiments of this invention the set of holes is generally coincident in spacing with the microfluidic channels. When a reflective beam splitter is used in the beamforming optics it must also match the holes.

While the simplest implementations use uniformly set channels and evenly set holes and possibly evenly set grooves in the beam division this is not required by the invention and similar embodiments may be designed for use with irregular spacing or channel patterns.

An alternative embodiment of the A7 fluorescence detection subsystem is to add narrowband pass filters before or after the fibers in the image plane (3-5), (2-8). A 400 micron fiber in this plane will capture a bandwidth of 10 nm. The addition of 10nm or 5nm bandpass filters will improve sensitivity and reduce noise in some cases. The present invention has been described with respect to an illustrative embodiment. Since certain changes may be made to the above constructions without departing from the scope of the invention, all subject matter contained in the above description or illustrated in the accompanying drawings should be construed as illustrative and not in a limiting sense. It is also understood that the following claims should cover all generic and specific features of the invention described herein, and all scope statements of the invention that are to be considered as included therein.

Claims (12)

1. Optical detection system (8) for observing a micro-fluidic system (10) containing a set of channels (3) for the transport of particles or molecules, characterized by the fact that it comprises: a light source (11) to produce a beam of light (4); a beam format optics assembly (12) for focusing the light beam (4); a set of holes (13), each hole (13a, 13b) combined with and associated with a micro fluid channel in said channel set (3) in the micro fluid system (10); at least one column detector strip (63) for receiving and carrying one of the optical extinction, advance spread and lateral spread produced after the beam of light passes through one of said channels through one of said holes; and a high numerical aperture fluorescence detector (17) receiving optical signals produced by a particle in any of said channels (3) when the particle forms an intersection with said light beam (4). 2. System for formatting a single incident beam (82) into a smaller beam array (84) with a controlled separation using a reflective beam splitter (80), characterized in that it comprises: a uniform set of reflective grooves; and wherein the set of reflective grooves is positioned relative to an incident beam at a selected angle. 3. Optical detection system (8) for interrogating a micro fluid system (10) including a set of micro fluid channels (3) on a particle-carrying substrate, characterized by the fact that it comprises: a light source (11) to produce a beam of light (14); a beamforming optics assembly (12) including a reflective beam splitter (80) for dividing the light beam (14) into a plurality of subsidiary light beams; and an optical mask (13) formed on the surface of the substrate, wherein the optical mask (13) includes a set of holes formed therein and coincident with a set of micro fluid channels in the micro fluid system (10), where the optics of beam formatting (12) direct each of said plurality of subsidiary light beams through one of said holes. 4. Optical detection system (8) for observing micro-fluidic systems (10) containing particle or molecule-carrying channels (3), characterized in that it comprises: a light source (11) for producing a beam of light (14) ); a beamforming optics assembly (12) for focusing the light beam (14); a set of holes (13) coincident with the micro fluid channels, wherein said set of beam-forming optics (12) passes the light beam (14) through said set of holes (13); at least one column detector strip (63) for receiving and carrying one of the optical extinction, advance spread or lateral spread produced after the beam of light passes through one of said channels (3) through one of said holes (13); and a high numerical aperture fluorescence detector (17) for simultaneous interrogation of a plurality of channels in the micro fluid system (10) and simultaneous detection of three fluorescence wavelength bands. 5. Optical detection system (8) for observing micro-fluidic systems (10) containing channels (3) carrying particles or molecules, CHARACTERIZED by the fact that it comprises: a light source (11) to produce a light beam ( 14); a beamforming optics assembly (12) for focusing the light beam (14); a hole in communication (13a, 13b) with a micro fluidic system channel; and a high numerical aperture fluorescence detector (17) for simultaneous interrogation of a plurality of channels in the micro fluid system (10) and using an image intensifier (220) as an optical amplifier element, wherein the amplifier of The image includes an input window (221) for an image signal, a light-sensitive electron emitter (222) for transforming light emitted from a photoelectron channel, an electron multiplier (223) for electron multiplication, a flourescent substance screen (224) for converting electrons into light and an exit window (225) for emitting an amplified image signal. 6. Optical detection system (8) for observing micro fluidic systems (10) containing channels (3) carrying particles or molecules, CHARACTERIZED by the fact that it comprises: one or more lasers (11) illuminating a set of spatially extended points through a mask (13) which is opened only at the points to form a first set of holes; a first high numeric aperture lens (45) positioned to capture light from the entire spatially extended set of points at one time; a spectral separation element for receiving and bending light from the first lens (45); a second high numerical aperture lens (47) for capturing light bent by the spectral separation element and imaging of the light captured in a second hole assembly; an image intensifier (220) behind the second hole assembly for detecting and amplifying light passing through the second hole assembly; and a photodiode detector array (15, 16) for capturing and transforming light from the image intensifier (220). Optical detection system according to claim 6, characterized in that it further comprises an electronic data acquisition system coupled to photodiode detectors (15, 16) to acquire an electronic output from photodiode detectors (15, 16 ). Optical detection system according to claim 6, characterized in that it further comprises a laser band blocking filter (74) disposed between the dot set and the image intensifier (220). Optical detection system according to claim 6, characterized in that the detector is capable of amplifying low intensity optical spectra having a duration of less than one millisecond from a spatially extended plurality of points and transforming the spectra. on electronic signals. 10. Optical system, characterized in that it comprises: a light source (11) for producing a beam of light (14) that passes through an object to be monitored; a first high numeric aperture lens (45) for capturing light from the light source (11); a scattering element (46) for receiving and bending light from the first lens (45); a second high numerical aperture lens (47) for capturing light bent by the scattering element (46); and a detector (50) including a light amplifying element for detecting a light signal from the second lens (47) and transforming the light signal into an electronic signal. Optical system according to claim 10, characterized in that the light amplifying element comprises a set of phototubes. Optical system according to claim 10, characterized in that the light amplifying element comprises an image intensifier-based multi-channel plate coupled to a photodiode detector array.