UV Raman microscope analysis system

Abstract

A method and system for interrogating a target for one or more chemical species of interest using Raman microscopy and spectroscopy. A feature includes the ability to precisely electro-mechanically move and orient a Raman microscope relative to the target with multiple degrees of freedom of movement, including targets with 3-D form factors. This promotes effective detection of minute quantities of chemical species of interest. It also allows effective detection of minute quantities whether the target is static or moving. The method and system can include enhancements. Examples include alternative imaging spectrometers, alternative Raman microscope optical set-ups, and alternative focusing techniques. Others include control of the excitation energy and user controls and options to allow highly adjustable, flexible, and effective detection for a variety of detection applications.

Claims

1. A method of detection of chemical species of interest on a target surface using Raman microscopy and spectroscopy comprising: a. interrogating a target for one or more chemical species of interest using a Raman microscope; and b. precisely electro-mechanically moving and orienting the Raman microscope relative to the target with multiple degrees of freedom of movement, including targets with 3-D form factors; c. further comprising selecting a type of focusing of the Raman microscope, wherein the type of focusing is one of: i. use of a range finder; ii. use of a visual channel comprising a quantification of blurriness in an acquired image related to sharpness of focus; and iii. use of a stereoscopic camera with overlapping field of view.

2. The method of claim 1 further comprising moving the Raman microscope into close proximity to target.

3. The method of claim 1 further comprising moving the Raman microscope across a surface of the target to scan the surface.

4. The method of claim 1 further comprising moving the target during the interrogation.

5. The method of claim 1 further comprising selecting a type of imaging spectroscopy for use with the Raman microscope.

6. The method of claim 1 further comprising selecting a type of optical set-up for the Raman microscope.

7. The method of claim 6 wherein the type of optical set up is selected from: a. a co-axial illumination of the target with collection of Raman scattering from the target; and b. an oblique illumination of the target with collection of Raman scattering along a different optical axis.

8. The method of claim 1 further comprising control of one or more of: a. on/off time of illumination by a laser for anti-particle ablation control; b. defocusing of illumination by a laser for anti-particle ablation control; c. scanning mode comprising one of automated or manual; d. display of field of view of the Raman microscope and other graphic or data content.

9. A Raman microscope analysis system comprising: a. an electro-mechanically controlled XYZ positioner; b. a Raman microscope adapted for collecting Raman scattering mounted on the XYZ positioner; c. an imaging spectrometer operatively optically connected to the Raman microscope for analyzing the collected Raman scattering for Raman content indicative of a chemical species of interest; d. a processor programmed to: i. control the XYZ positioner to adjust the Raman microscope and its orientation into proximity to and relative a target object; ii. illuminate the target with excitation illumination; iii. collect Raman scattering; iv. evaluate the collected Raman scattering for a chemical species of interest; and e. a user interface to select modes of operation of the system.

10. The system of claim 9 wherein the electro-mechanically controlled XYZ positioner comprises a 6-axis articulated arm.

11. The system of claim 9 wherein the Raman microscope comprises: a. a UV laser as an illumination source; b. a microscope objective lens; c. optics to direct collected Raman scattering to the imaging spectrometer.

12. The system of claim 9 wherein the imaging spectrometer comprises a CCD imager.

13. The system of claim 9 further comprising an interrogation space within the reach of the XYZ positioner.

14. The system of claim 13 further comprising one of: a. a platform for supporting a target object in the interrogation space; and b. a conveyor for moving a target object past the interrogation space.

15. The system of claim 9 wherein the Raman microscope comprises one of: a. a co-axial illumination of the target with collection of Raman scattering from the target; and b. an oblique illumination of the target with collection of Raman scattering along a different optical axis.

16. The system of claim 9 wherein the Raman microscope comprises a type of focusing from one of: a. a range finder; b. a visual channel comprising a quantification of blurriness in an acquired image related to sharpness of focus; and c. a stereoscopic camera with overlapping field of view.

17. The system of claim 9 wherein control of the excitation illumination comprises one or more of: a. on/off time of illumination by a laser for anti-particle ablation control; b. defocusing of illumination by a laser for anti-particle ablation control; c. scanning mode comprising one of automated or manual.

18. The system of claim 9 wherein the user interface includes a display of field of view of the Raman microscope and other graphic or data content.

Description

III. BRIEF DESCRIPTION OF THE DRAWINGS

(1) The patent or application file contains at least one drawing or photograph executed in color. Copies of this patent or patent application publication with color drawings(s) will be provided by the Office upon request and payment of the necessary fee.

(2) FIG. 1A is a diagrammatic view of a prior art stationary Raman spectroscopy system that analyzes a specimen (here a human hand) that is movable relative to the static system. This system is from U.S. Pat. No. 7,692,776 B2, issued Apr. 6, 2010, and incorporated by reference herein as background information.

(3) FIG. 1B is a perspective view of a portable UV Raman system that can be manually moved relative to a static target (here a duffle bag). This system is co-owned by the owner of the present application and found at co-pending U.S. Ser. No. 16/151,682, filed Oct. 4, 2018, and incorporated by reference herein.

(4) FIG. 2 is a perspective view of a system according to an exemplary embodiment of the present invention. The UV Raman system and its target (a duffle bag handle) are enlarged in the detail view.

(5) FIG. 3 is an example of a consolidated display from a scan of a target by the system of FIG. 2, including indication of detection of Potassium Nitrate powder or traces (KNO3) on the target, microscope images of the target, and other details about the detection. In this example of proof of concept, the data was taken on a UV Raman microscope setup such as FIG. 2 showing detection of a 130 nanogram (ng) KNO3 particle.

(6) FIG. 4A is a diagrammatic view of a first specific exemplary embodiment according to the invention (sometimes called version 1), namely a diagram of the optical design for what will be called the coaxial laser illumination approach.

(7) FIG. 4B is a diagrammatic view of a second specific exemplary embodiment according to the invention (sometimes called version 2), namely a diagram of what will be called a Scanning Raman Microscope.

(8) FIG. 4C is a diagrammatic view of a third specific exemplary embodiment according to the invention (sometimes called Approach 2: Oblique UV Laser Illumination),

(9) FIG. 5 is a graph and data providing a representative range vs. position plot with a curve fitted characteristic equation used with an optional range-finder-based focusing technique that can be used according to aspects of exemplary embodiments of the invention.

(10) FIG. 6 is a photograph illustrating an alternative optional visual focusing technique that can be used according to aspects of exemplary embodiment of the invention; here illustrating quantification of what is called a blurriness score when imaging a target, here showing a low blurriness score which indicates lack of focus.

(11) FIG. 7 is a photograph illustrating the focusing technique via blurriness score as in FIG. 6; here showing a high blurriness score which indicates good focus.

(12) FIG. 8 is, for comparison, side-by-side stereoscopic output contemporaneous-acquired images of the same target. The left-side image is a normal photograph of the target and background scene. The right-side image can be used to show depth of field. Brighter white objects are closer than darker objects.

(13) FIG. 9 is, for comparison, adjacent highly magnified acquired images that demonstrate features according to aspects of the invention; namely (top) a defocused laser of size roughly 400 um750 um, and (bottom) a focused laser of size roughly 250 um80 um.

(14) FIG. 10 is examples of screen displays of results from use of methods and systems according to the present invention, namely (top) what will be called an Auto Mode with Overlay Result Feedback and (bottom) what will be called a Spectra Plot acquired from a target.

(15) FIG. 11 is an example of a screen display useful with methods and systems according to aspects of the present invention, namely (left side) what will be called a Manual Mode with (right side) what will be called a Touchscreen Input Bounding Box.

IV. DETAILED DESCRIPTION OF EXEMPLARY EMBODIMENTS OF THE INVENTION

A. Overview

(16) For a better understanding of the invention, non-limiting examples of several different forms and embodiments the invention can take will now be described in detail. It is to be understood that these examples are neither exclusive nor inclusive of all forms and embodiments possible with the invention.

(17) For example, exemplary embodiments discussed below will focus upon detection of explosive such as potassium nitrate (KNO3) as a chemical species of interest. Those skilled in the art understand that the invention can be applied to any number of other chemical species for any number of purposes so long as they can be extracted from Raman scattering. For example, there can be other chemical species of interest to be detected relative to explosives for public, private, security, or military purposes. But there can be non-explosive chemical species of interest that could benefit from aspects of the invention. One example is detection of a pesticide or insecticide on fruit or vegetables.

(18) By further example, exemplary embodiments utilize a particular UV laser as an excitation source. As known to those skilled in the art, a variety of excitation sources are possible. Still further, exemplary embodiments utilize an excitation source that is designed to generate Raman scattering from the target, and isolate an effective Raman signal from a collected portion of that scattering for identifying chemical species of interest with sufficient accuracy, precision, reliability per target and repeatable for a substantial number of targets over a substantial useful life.

B. Generalized Embodiment

(19) With particular reference to FIGS. 2 and 3, a UV Raman microscope analysis system 20 according to one embodiment of the invention is illustrated. In this example, an integrated machine includes a Raman microscope subassembly 24 which can generate both an excitation source beam 25 focused to point 28 on target 29 as well as collect Raman scattering 27 (similar to 15 and 17 of FIG. 1B). A platform or receiver is presented on which a target 29 can be supported with at least most of its unsupported sides exposed. A shroud or barrier 39 can be adjustable to surround and enclose target 29 and Raman microscope 24 during operation for eye safety.

(20) We researched utilizing a UV Raman microscope to enable low level detection (nanograms) of explosive particles. An overall system concept is shown in FIG. 2.

(21) 1. Raman Microscope Scanning Method

(22) Classical Raman microscopes are designed to look like a standard optical microscope in which the sample is moved relative to the instrument. This works well for flat samples. However, in this embodiment, the scanning mechanism is for the optical instrument to move relative to the sample with the use of an articulated arm as shown in FIG. 2. The advantages of this approach include at least: a. It allows for the sample to either be stationary or moving at a slow rate of speed. b. It allows for scanning of irregular 3-D surfaces as most real-world objects are. c. It allows for access to a larger surface are or portion of the sample than a standard line-of-sight optical system.

(23) 2. Use of Non-Spatial Raman Imaging for Fluorescence Rejection:

(24) The owner of the present application has previously disclosed the details of the concept (application U.S. Ser. No. 16/151,682 filed Oct. 4, 2018, incorporated by reference for background information). Embodiments of the present invention can optionally employ that concept to further minimize fluorescence interference. FIG. 3 shows detection of a 130 ng particle of Potassium Nitrate (KNO3) on black plastics ABS (a fluorescent substrate). Without this approach the key signal corresponding to the KNO3 would not be detectable. But other Raman set-ups can be used.

(25) FIG. 3 shows proof-of-concept data taken on a UV Raman microscope setup showing detection of a 130 ng KNO3 particle.

(26) Further aspects of the generalized embodiment follow.

(27) The platform or receiver to support a target 29 can be static (e.g. a tabletop, a bench, a surface, or the like). However, it might also be a conveyor (see diagrammatic depiction in FIG. 4A). A conveyor could move a target, or a series of successive different targets, past an interrogation zone. It could be a conveyor belt that is synchronized with acquisition of Raman signal from each target as it moves by the interrogation zone.

(28) System 20 includes a user interface with a display 22. It can include a keyboard, touch screen, or other user controls. It also includes support equipment 30. In an enclosure box, as indicated in FIG. 2, support equipment 30 can include a spectrometer 60, processor 34, laser driver 36, motion controller 46, and object tracker 54.

(29) It is to be understood that spectrometer 60 could be as is disclosed in detail in any of its embodiments in incorporated by reference co-pending co-owned Ser. No. 16/151,682 or other.

(30) As indicated in FIG. 2, Raman microscope 24 is at the distal end of robotic arm 40 that has six-axis articulation or degree of freedom of movement (DFOM). A connection 43 at distal section 42 of arm 40 allows Raman microscope to be electromechanically moved in almost any aiming orientation for beam and collection field of view 25/27 relative to target 29, at least as to surfaces of target 29 that are exposed and not on a supporting surface. As can be appreciated, alternatively, there could be some sort of suspension system to suspend 3-D target 29 so that even under surfaces could be reached by articulated arm 40. Proximal end 44 of arm 40 is mounted on a mount 45 that is fixed relative to the station 20.

(31) As can be appreciated by those skilled in the art, robotic arm 40 can be controlled in a number of ways. In this embodiment, machine vision subsystem 48, with a digital camera 47 with field of view over the range of relevant positions within barrier 39, could track the distal end of arm 40 and Raman microscope 24 in 3-D space, convert its coordinates to camera space, and inform motion controller 46 and processor 34 of actual physical location in real space. This could allow a human operator at user interface 22 to manually control scanning of the excitation beam 25 relative to target 29. It also could allow, through appropriate machine vision algorithms and software, for there to be an automated scanning of many if not all exposed surfaces of target 29. Furthermore, by appropriate machine vision algorithms, image recognition could be used for the machine vision system to recognize preprogrammed shapes or features of a target and only scan or focus in unrecognized preprogrammed such features.

(32) It can also automatically sense distance from any target 29 within the interrogation zone and the reach of arm 40. This can be by one or more proximity sensors on or associated with arm 40 or the interrogation zone. It could be by a component such as a range finder. It could be by evaluation of imaging of the target (e.g. use of depth information in acquired digital images). Proximity sensing allows automatic or semi-automatic control of how close Raman microscope 24 is relative to any surface of target 29, including down to centimeter and even millimeter scale. This can enhance the ability of the system to focus to very small areas of the target surface and acquire highly magnified field-of-view images that can be effectively evaluated for Raman content.

(33) Such machine vision systems, motion controllers, object trackers, and algorithms for machine vision and motion control are available from a wide variety of commercial sources.

(34) Again, in this embodiment, Raman microscope 24 can be a version as disclosed in incorporated by reference U.S. Ser. No. 16/151,682. As disclosed therein, it uses a technique to obtain high resolution of very small physical space areas to help get good Raman signal for small areas. As such, utilizing the same can enhance and promote microscopic detection of chemical species of interest. This can include micro scale and even nano scale. By using appropriate optics, collected Raman scattering can be digitally imaged at a resolution that helps, on a pixel by pixel basis, isolate very small regions of the field of view from which Raman scattering is collected towards this purpose. It is to be understood, however, that alternative Raman microscopes can be used.

(35) Again, utilizing the above features could allow an efficient and effective interrogation of all exposed surfaces of a three-dimensional target automatically. As can be appreciated, alternatively, just certain portions of a target could be scanned. One example might be, as shown in FIG. 2, just the handle of a duffel bag. This might have a higher probability of containing minute (even nano scale) particles related to explosives such as might be transferred from the handler of the explosive hand to the handle when the duffel bag is carried.

(36) Furthermore, system 20 could allow an almost fully automated scanning of a target including three-dimensional target without a human user having to reposition either the target or the Raman microscope.

(37) As indicated in FIG. 2, a rough analogue would be a medical imager such as an Mill or CT scanner. Programming can intelligently manipulate the aiming direction field of view over 3-D surfaces. This would allow barrier 39 to cover the entire target during a scan for safety reasons. Additionally, personnel could multitask to other tasks while the scanning is occurring. Programming could, however, issue an alarm and stop the scanning process immediately upon an indication of a chemical species of interest. The designer could select how the scanning proceeds.

(38) FIG. 3 gives one example of a possible display at the user interface 22. In this example, related to scanning for potassium nitrate (KNO3), a chemical species that could be related to explosives, during the scan a user interface could include at displayed content 50, a section that shows concentration in the spectral area related to KNO3. See in window 51 reference numeral 52 and various indicated levels of KNO3 at 52A, B, and C related to the number of pixels based on bundled optical fibers according to the detection process of co-pending Ser. No. 16/151,682. As can be seen, the more optical fibers utilize the higher the intensity of the Raman signal for the spectra related to KNO3. As such, the higher the intensity the better the ability to distinguish it from noise or irrelevant content added around that spectral band. Details of this proof-of-concept results are as follows.

(39) In the example of FIG. 3, based on an actual test where the sample included KNO3 (potassium nitrate) the duffel bag substrate is ABS, the distance between explosive particles. The focal distances at or approximately 1 inch, KNO3 particles on the order of a diameter of 52 m and on a mass on the order of 133 nanograms were detected at 52A-C over an interval time of 500 ms. As such, this proof of concept shows that the combination of being able to position the Raman microscope close to but offset from any orientation on an exposed 3-D surface was capable of detecting at a nano scale level a chemical species of interest.

(40) FIG. 3 also shows that if configured accordingly, user display 22 could also show spectral hyper-cube 53, a sample actual greatly magnified image of the sampling area, including an area 54, but also it could show optionally with graphics (here an overlaid red circle) the specific focus of interrogation for a single micro or nano scale particle 55A (of many such possible particles 55B-H). This could help a user see the nature of what is being scanned and detected and help quality control by distinguishing it from, for example, background 57 or other materials 56. FIG. 3 also shows that data 58 relating to the scan could also be displayed. This shows that according to the algorithm used, specific information about an indication of a correct classification of a chemical species of interest (here an explosive threat) was identified. This could be documented and stored as for evidentiary purposes.

(41) It can therefore be seen that the invention achieves at least one or more of its stated objectives. Balancing competing factors such as cost, complexity, speed of detection, accuracy of detection, safety of humans, and the like, the system 20 improves over bench top systems that are static relative to excitation aiming and field of view and require relatively flat surfaces to get any significant chance of accurate and reliable detection. It also takes away some human error when having hand-held manual aiming of the microscope. It furthermore has been shown to have capabilities of effectively and efficiently interrogating a wide variety of targets including those of substantial 3-D form factor with detection of micro and nano scale traces of chemical species of interest. As such, this could be highly beneficial for effective and efficient semi-automated or automated screening such as for security purposes.

C. Enhancement Options

(42) The generalized system of FIGS. 2 and 3 could include some optional enhancements. A first example is discussed below.

(43) 1. Optical Design Details

(44) Two optical designs can be used with the system 20, each of which has advantages and disadvantages. They are described below.

(45) In this optional optical set-up for generalized embodiment 20, the UV Raman microscope 24 utilizes a reflective objective with a 25 mm working distance to the target. The output is collimated and spectrally split using dichroic optical elements. A visible portion is directed to an operator camera with its own focus lens to provide a highly magnified view of the target area. The Raman band is directed to a fiber bundle at the focus of another lens.

(46) Approach 1: Coaxial UV Laser Illumination Set-Ups 60A and 60B

(47) With particular reference to FIGS. 4A and B, a laser source 68 generates a 262 nm deep UV laser 67 which is collimated and injected into the objective 74 optical axis with a long pass dichroic 66. The laser is further reflected by a long pass dichroic 72 into the reflective objective 74 (see FIGS. 4A and B, versions 1 and 2). The objective 74 is focused at the target 29 which results in a laser spot 28 measuring a few tens of microns in diameter. It is to be understood other UV wavelengths are possible, for example, UV wavelengths in the approximate range of 220-266 nm are envisioned. See FIG. 4A. The main difference between the set-ups of FIGS. 4A and B is that set-up 60A of FIG. 4A uses an imaging spectrometer and ICCD (integrated charge-coupled device) 62A as in U.S. Ser. No. 16/151,682, whereas set-up 60B of FIG. 4B uses a different spectrometer and imager. As will be understood, principles of the generalized embodiment and this optional enhancing feature can be applied to different types of imaging spectrometers.

(48) Further details about set-ups 60A and B follow.

(49) With specific reference to FIG. 4A, a specific optical set up for Raman microscope 24 is shown at 60A. An imaging spectrometer and ICCD (integrated charge-coupled device) using the circular to linear fiber-optic arrangement of co-pending Ser. No. 16/151,682, could be optically connected via cabling as shown in FIG. 4A. The bundle optical fiber cable from 62A would collect return electromagnetic energy at 65 that would be focused on those bundled ends 63 by focusing lens 64. Laser beam 67 would excite Raman scattering by directing beam 67 from deep UV laser source 68 to dicrotic mirror 66. It would move at 69 to long pass dicrotic 72 and into focusing lens 74. As such deep UV Raman 73 would be excited via the focus beam 25 on a small area of 28 of target 29. As shown, in this example the robotic arm would position the interrogation beam close to spot 28 on target 29 regardless of orientation of that handle in 3-D space.

(50) The return scattering 27 would pass into the field of view of lens 74, go back at 73 and reflect off dicrotic 72 at 69 and 65 into lens 64 for focusing on the ends of fiber-optic cable 62A for imaging and Raman content extraction.

(51) Note, moreover, that this set up also allows visible light in the field of view of lens 74 to pass through dicrotic 72 at visible wavelengths at 81 and through lens 82 at 83 to visual band camera 84 (see subsystem 80 of FIG. 4A). This could produce the sample image 54 in FIG. 3 for human viewing or storage for documentation or quality control or for other purposes as disclosed herein.

(52) FIG. 4B is a slight alternative to FIG. 4A with the same or similar components of FIG. 4A, except for the spectrometer. Any effective spectrometer 62B with fiber coupling could be used with a similar set up to FIG. 4A. A nominally collimated deep UV laser beam 67 can be used. This similarly could use the robotic arm for beneficial positioning close to the target 29 as well as provide a visual band recordation of any part of the scan.

(53) Approach 2: Oblique UV Laser Illumination Set-Up 60C

(54) Alternatively, in the set-up 60C as in FIG. 4C, the 262 nm laser 67 from laser generator 68 is collimated and projects the beam 67 onto the target 29 at the closest possible angle to the microscope axis. A discrete focus lens is the final element in the laser path. The lens is mounted to a focus adjust stage (not shown but commercially available) which provides a means to vary the size of focus spot 28 on the target 29 (see FIG. 3C). Laser line rejection filter 66 can replace dichroic 66 of FIG. 4A because of direct aiming of laser 67 to target 29. Other components of 60C can be the same or similar to 60A or 60B. Return scattering 69 is passed back to spectrometer 62A (or 62B).

(55) Relative Advantages of the Above-Approaches

(56) The coaxial focus (60A or 60B of FIG. 4A or B respectively) provides a tight focus spot 28 driven by the f number of the objective 74. This can be used to limit background illumination fluorescence. As the objective is an axis design, the laser will lose energy from the secondary obscuration.

(57) The direct focus layout 60C of FIG. 4C eliminates a dichroic from the Raman optical path which improves throughput due to coating inefficiency. It also provides a means to vary spot 28 size on the target 29 as a means to reduce irradiance level which can ablate and degrade the target signature. The spot size on the target depends on the f number of lens which in turn must accommodate physical clearances created by the reflective objective 74. An optimized design would attempt to match the f number of the objective 74 which will require customized fixtures to hold the focus lens as close as possible to objective housing.

(58) Thus, FIG. 4C is a still further alternative for the optical set up 60C according to the generalized invention. It is basically a similar set up to FIG. 4A with the following major difference. Instead of the excitation beam being injected into the optical path through a dicrotic mirror, it is directly aimed at the focusing point of optic 74.

(59) Unique Focusing Approaches

(60) Ensuring the Raman channel is in focus for a given sample is vital to optimizing return signal in Raman Microscopy. Various techniques that could be used with the generalized embodiment 20 are listed below 1) Use of a Standard Rangefinder a) A standard rangefinder (commercially available and diagrammatically indicated in FIG. 4C) can provide accurate measurements with millimeter level precision. Using this range, a characteristic equation 90 can be developed to precisely move the focusing optics into position (see FIG. 5). For example, actual position data 91 can be collected. Range finder readings can be correlated to robotic arm 40 positions 94 (e.g. by recording/extracting in coder counts for motion controller 46 which has the electrical motors or actuators that move arm 40. The equations/curve fittings 92/93 thus extrapolate a continuum of ranges correlated to arm position that can be used for actual focusing. For any given range, each focusing stage will have a unique position that will be predetermined. Such a range finder would be operatively connected to the system processor to inform it of distance between the distal end of objective 74 and the target 29. The characteristic developed from the curves of FIG. 5 can then be used to move focusing components of the system. 2) Use of the visual channel

(61) With reference to FIGS. 6-7 an optional alternative to focusing is illustrated. Its operation would be as follows. a) Through the visual channel (e.g. using visual band camera subsystem 80), blurriness of an acquired image can be observed. Blurriness is generally subjective, but prior research has shown that by using a variation of the Laplacian, a relative number can be produced to represent blurriness of an image. Maximizing this value as the focusing optics move can generate a focused visual channel which would correspond to a focused Raman channel (see FIGS. 6 and 7). This is performed by: (1) initially focus the optics of the system (FIG. 6, 100A); (2) perform variance of the Laplacian on the image to get a blurriness score (e.g. 2.75 of FIG. 6), where the smaller the score the blurrier and less focused and the higher the score the sharper and more focused. (a) which can be done by convolving a single image channel with the Laplacian kernel, and then computing the variance of the result; (3) move the focus optics in one direction; (4) determine if blurriness score increased or decreased; (5) If increased (e.g. FIG. 7 at 100B), continue moving in that direction until it decreases; (6) if decreased, move in opposite direction until blurriness score decreases again; (7) return to the position which the blurriness score was optimized.

(62) In mathematics, the Laplace operator or Laplacian is a differential operator given by the divergence of the gradient of a function on Euclidean space. It is usually denoted by the symbols .Math., .sup.2 (where is the nabla operator) or . The Laplacian .Math.f(p) of a function fat a point p, is (up to a factor) the rate at which the average value off over spheres centered at p deviates from f(p) as the radius of the sphere shrinks towards 0. In a Cartesian coordinate system, the Laplacian is given by the sum of second partial derivatives of the function with respect to each independent variable. In other coordinate systems such as cylindrical and spherical coordinates, the Laplacian also has a useful form. Thus, this generation of a value related to blurriness can be used to enhance focusing of the system.

(63) As will be appreciated, an automation of focus could be achieved. The system 20 could be automatically controlled to train or learn optimized focus for a given target location. By this hunt and peck technique enabled by the ability to move the Raman microscope electro-mechanically relative the target over minute (e.g. micrometer scale) increments, it could self-determine and optimize sharpness of image and take Raman measurement there. As uch this could enhance accuracy, precision, and efficiency of detection.

(64) 3) Use of a Stereoscopic Camera a) Using a visual band stereoscopic camera (e.g. camera 84 of subsystem 80) (commercially available), depth of field can be determined. These cameras are a combination of two cameras with predetermined specifications that look at an overlapping field of view. Depth is determined by utilizing the cameras' focal lengths, distance between the two cameras, pixel size, and difference in pixel position between the two camera images. Using this depth, a characteristic equation can be developed to precisely move the focusing optics into position. For any given depth, each focusing stage will have a unique position that will be predetermined. i) Depth is calculated based on the focal length of the two cameras, distance between the two cameras (see stereogram 110 of FIG. 8). As shown in FIG. 8, brighter objects 115A/115B are closer than darker objects 113A/113B or 114A/114B in left and right images 112A and 112B of FIG. 8.

(65) 3. Anti-Particle Ablation Control

(66) Using a powerful, the focused laser beam 25/27 of system 20 comes with the need to control the amount of energy on samples (e.g. chemical species of interest) and substrates (e.g. the small target area(s) 28 on target(s) 29) to prevent ablation and damage. This is particularly important for interrogating sensitive samples that easily evaporate or burn off or interrogating samples that reside on expensive or important substrates like an expensive handbag. Techniques that can be used are listed below: 1) Controlling Laser On/Off time a) Custom firmware can precisely control when the laser turns on, and how long it stays on. The laser of system 20 can be controlled with 10 ms precision with capability of control down to 1 ms precision. As shown in Equations (1)-(3) below, energy density functions multiplicatively in relation to laser on time so, by reducing the amount of time the laser is on, a sample by 10x will result in a 10 reduction in energy density.
A.sub.circle=(/4)(diameter).sup.2(eq. 1)
Power density (watts/m.sup.2)=laser power (watts)/area.sub.circle(eq. 2)
Energy density (J/m.sup.2)=power density (watts/m.sup.2)dwell time (s)(eq. 3)
As such, using this technique can enhance and be beneficial in effectiveness of a system 20. 2) Reducing Power Density by defocusing the laser a) Focusing optics of system 20 are designed in such a way to allow for independent control of both the transmit and receive channels. This allows the receive Raman channel (the one which collects the Raman scattering for processing out of it a Raman signal) to remain in optimal focus while allowing the transmit laser (the excitation energy to the target 29) to be defocused to prevent ablation. As shown in the equation (2) above, if Area.sub.(circle) increases, power density decreases. This in turn causes energy density to also decrease. FIG. 9 shows the same laser beam in two different focus configurations. Although the overall output power of the laser is the same, the energy density of the bottom focused laser (see image 124 in FIG. 9) is nearly 12 that of the top defocused laser (see image 122 in FIG. 89).

(67) 4. Novel Control and Operator Feedback for Raman Microscope Chemical Detection:

(68) The Raman Microscope Chemical Detection System disclosed herein provides more intuitive control and feedback via a large touchscreen display (see, e.g., display 22 of FIG. 2). It has advanced antiparticle ablation controls including laser on/off time and the ability to defocusing of the beam to reduce power density. See example display screens 130 and 150 of FIGS. 10 and 11). This stands out from state-of-the-art systems. The Raman Microscope Chemical Detection system disclosed herein can contain automatic scanning mode 131 and manual scanning mode 151 (FIGS. 10 & 11). In automatic scanning mode 131 of FIG. 10, the software application uses object recognition algorithm to identify in images 134 and 136 targeted area 135. In manual mode 151 of FIG. 11, the user may select the desired view 152 of the object and draw the bounding box 155 of the area to be scanned via touchscreen 22. The software application used with the system can provide the user with multiple views (134 and 136 of FIGS. 10 and 152 and 156 of FIG. 11) of the targeted object using wide angle cameras that covered 360 plus the bird's eye view. The application can display live feedback while the scanning process is in progress by distinctively overlaying results data over an image of the scanned object. Here, the overlays consist of a series of red/green squares 137 and 138 or 157 and 158 indicating threat/non-threat areas respectively. A user may zoom in on this overlay with a touchscreen gesture and tap on a particular square to access result details 140 and spectra plots (e.g. 142/142A) for that particular region and for textual information (e.g. 133, 143). The results overlay image can also be saved to the hard drive for future reference.

(69) To summarize, the unique features of Raman Microscope Chemical Detection system and application disclosed herein, which can be used individually or in any combination, include at least the following: 1) Antiparticle ablation controls a. Laser on/off time b. Defocusing of the beam to reduce power density 2) Dual scanning mode, automatic and manual 3) Draw input bounding box for scanning via touchscreen 4) Full 360 view plus bird eye view of targeting object 5) Overlay object image with result feedback (display live and save to hard drive) 6) Access detailed information and spectra plots from image result. Other feedback, information, or results can be displayed. See, e.g., ref. nos. 132, 133, 140, 142, 143 and 156.

D. Options and Alternatives

(70) As emphasized above, the exemplary embodiments and their aspects are just a few examples of forms the invention and its aspects can take. For example, variations obvious to those skilled in this technical art will be included within the invention.