HK40006971B - Apparatus and method for sample processing for microscopy - Google Patents

Description

Apparatus and method for sample processing for microscopes

Cross Reference to Related Applications

Priority of united states provisional patent application No. 62/320120 filed 2016, 4, 8, 35 u.s.c. § 120. This application is incorporated by reference herein in its entirety.

This application relates to U.S. patent application No. 15/066065 filed on 10/3/2016; U.S. patent application No. 62/131164 filed on 3/10/2015; U.S. patent application No. 14/314743 filed 24/6/2015; us patent application No. 61/839735 filed on 26.6.2013; U.S. patent application No. 14/173500 filed on 5/2/2014; U.S. patent application No. 61/255781 filed on day 10/28 of 2009; U.S. patent application No. 12/913639 filed on day 27 of 10/2010; united states patent application No. 13/095175 filed on day 27 of month 4, 2011; us patent application No. 61/761467 filed on 6.2.2013 and us patent application No. 61/785762 filed on 14.3.2013. These applications are incorporated by reference herein in their entirety.

Technical Field

The present disclosure relates to sample processing for microscopes.

Background

In a typical optical microscope, light passing through a sample is transmitted through a lens to the user's eye, film or sensor, and then an image representative of the sample is formed.

In other approaches, light representative of the sample may be detected without a lens and used to form an image of the sample by placing the sample on or near a detector (e.g., an integrated circuit) that includes an arrangement of photosensitive elements. The signals generated by the detector may be processed to derive an image.

Disclosure of Invention

In one aspect, an apparatus may comprise: a light sensitive imaging sensor configured to receive a fluid sample on top of a surface of the light sensitive sensor; a body configured to move relative to a light sensitive imaging sensor; and a carrier device configured to move the surface of the body relative to the light sensitive imaging sensor such that, when the surface of the body contacts a portion of the fluid, the surface of the body (i) is substantially parallel to the surface of the light sensitive imaging sensor and (ii) settles on the fluid sample independent of the motion of the carrier.

In some embodiments, the body allows light to pass onto the light sensitive imaging sensor.

In some embodiments, the surface of the light sensitive imaging sensor for receiving a fluid sample comprises a hydrophilic coating.

In some embodiments, a surface of a portion of the body that contacts the fluid sample comprises a hydrophilic coating.

In some embodiments, the device further comprises a sample delivery component for preparing and delivering a fluid sample to the surface of the light sensitive imaging sensor. In some cases, the sample transport component comprises at least two volume capillaries, a nozzle for mixing fluids within the at least two volume capillaries, and an output tip through which a fluid sample is transported to a surface of the photosensitive imaging sensor.

In some embodiments, the body includes an extension on the carrier. In some cases, the extension of the body has features that mate with corresponding features on the carrier.

In some embodiments, the apparatus further comprises means for adjusting a vertical distance between a bottom surface of the carrier and a surface of the light sensitive imaging sensor receiving the sample fluid.

In another aspect, a method may include: a body is moved toward a fluid sample on a surface of a light sensitive imaging sensor such that when the surface of the body contacts the fluid sample, the surface of the body is parallel to the surface of the light sensitive imaging sensor and the body settles on the fluid sample.

In some embodiments, moving the body toward the fluid sample comprises placing the body on a carrier such that a center of the body is vertically aligned with a center of the light sensitive imaging sensor.

In some embodiments, moving the body toward the fluid sample comprises moving the carrier toward the fluid sample.

In another aspect, an apparatus comprises: a solid member; a photosensitive imaging sensor; a deformable member coupling the solid member and a surface comprising the light sensitive imaging sensor, the deformable member comprising a sidewall enclosing a fluid chamber configured to receive a volume of fluid, the surface of the fluid chamber comprising a light sensitive chamber; and means for deforming the deformable member so as to adjust the height of the fluid chamber.

In some embodiments, the solid member allows light to pass into the fluid chamber.

In some embodiments, the means for deforming the deformable member comprises a transparent top pressurizable chamber enclosing the fluid chamber.

In some embodiments, the base comprises an integrated circuit board.

In another aspect, a method comprises: injecting a fluid sample into the chamber; deforming a deformable member to reduce the volume of the chamber to reduce the volume of the sample; and capturing an image of a portion of the sample at a photosensitive sensor surface within the chamber after reducing the volume of the sample.

In some embodiments, the method further comprises deforming the deformable member to increase the volume of the chamber.

In some embodiments, deforming the deformable member to reduce the volume of the chamber includes drawing a volume of gas within an expandable chamber within the chamber.

In another aspect, a point-of-care device, comprising: the sample processing chamber includes: a base having a chamber and a light sensitive sensor having a surface within the chamber that receives a fluid sample, and a body that moves relative to the light sensitive imaging sensor and has a surface that contacts a portion of the fluid sample such that when the surface of the body contacts a portion of the fluid, the surface of the body (i) is parallel to the surface of the light sensitive imaging sensor, and (ii) settles on the fluid sample; a device coupler for electronically coupling to a mobile device capable of accepting electronic communications corresponding to signals derived from the light sensitive imaging sensor; and a housing for holding the sample processing chamber and device coupler.

In some embodiments, the surface of the light sensitive imaging sensor for receiving a fluid sample comprises a hydrophilic coating.

In some embodiments, the point-of-care device includes a sample delivery component for preparing and delivering a fluid sample to a surface of the light-sensitive imaging sensor.

In some embodiments, the sample transport component comprises at least two volume capillaries, a nozzle for mixing fluids within the at least two volume capillaries, and an output tip through which a fluid sample is transported to the surface of the photosensitive imaging sensor.

In some embodiments, a surface of the body that contacts a portion of the fluid sample is on a component that is separable from the body.

In some embodiments, the component separable from the body comprises a plate and a protruding element that is lowered into a recessed chamber of the base.

In some embodiments, the top surface of the protruding element is the same size as the surface of the portion of the body that contacts the fluid sample, and the top surface of the protruding element is the surface of the portion of the body that contacts the fluid sample.

In some embodiments, the shape of the protruding element comprises a truncated pyramid.

In some embodiments, the electronic communication exchanged between the mobile device and the light sensitive imaging sensor comprises instructions to capture an image of a portion of a fluid sample placed on a surface receiving the fluid sample.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In the event of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

The details of one or more implementations are set forth in the accompanying drawings and the description below. Other potential features and advantages will become apparent from the description, the drawings, and the claims.

Drawings

Fig. 1 is a schematic diagram showing an example of a contact microscope system.

Fig. 2 is a cross-sectional view showing an example of lowering the top of the chamber onto the sensor surface of the contact microscope system.

Fig. 3A-3B are schematic diagrams illustrating an example of a contact microscope system in which the chamber top is lowered along one side.

Fig. 4A-4C are schematic diagrams illustrating techniques and components for dispensing a sample onto a sensor surface.

Fig. 5A-5B are schematic diagrams illustrating an example of an open chamber contact microscope system.

Fig. 6A-6E are schematic diagrams illustrating components of an open chamber contact microscope system.

Figures 7A-7C are schematic perspective views illustrating an example of a point-of-care microscope system.

Fig. 8 is a schematic perspective view showing an example of an improved closing mechanism for a contact microscope system.

Fig. 9A-9D show schematic diagrams representing examples of closed chamber contact microscope systems.

Detailed Description

Images captured using contact microscopy often require a well-defined contact surface between the photosensitive sensor surface and the particles to be analyzed. For quantitative techniques, the ability to calculate the exact number of particles in a fluid sample is based on the formation of a thin layer of the sample that is uniformly distributed over the sensor surface, where the height of the thin layer is approximately the diameter of the particle (e.g., a monolayer). However, due to various complexities in the microenvironment (e.g., fluid-surface interactions, lack of sufficient accuracy in adjusting the placement of physical components), establishing and maintaining a uniformly distributed sample layer prior to performing the imaging process is often difficult, complicating efforts to accurately reuse similar techniques in subsequent imaging processes.

One area in which contact microscopy techniques can be applied is in blood counting, where cells or cellular components such as red blood cells and platelets are counted in a carefully controlled volume of blood. Blood counts can be used to diagnose pathological and health conditions, determine the severity associated with such diagnoses, and monitor patients for changes in diseased conditions.

However, while these technologies are ubiquitous in health care systems in developed countries, their application is limited in developing countries. For example, blood counts can be expensive to manage and tend to be performed on dedicated machines operating in dedicated laboratories, such as in hospitals or clinics, which prevents their use in resource-limited or remote areas, where the lack of skilled operators often precludes the use of large-scale techniques with relatively high complexity.

Accordingly, the innovative aspects described throughout this specification relate to improving sample processing for contact microscopy techniques used to calculate blood counts, as well as other applications. The systems and techniques described herein provide a cost-effective method to improve the repeatability and accuracy of performing blood counts. For example, the structure of the system can be designed to enhance the techniques for establishing and maintaining a thin sample layer uniformly distributed over the sensor surface to consistently calculate cell counts. The figures and elements shown therein are not always drawn to scale and many of them are schematically shown. The spatial relationships of the elements in the figures may appear different than described in the text, e.g., above and below and top and bottom may be shown in the figures in reverse of the manner they are described in the text.

As used herein, "photosensitive location" includes any feature of the device, including photosensitive elements or pixels and light source locations, for example, that is sensitive to light alone or that is capable of emitting light, respectively, or both. The phrase light source position may refer to an element capable of emitting light. In some cases, the phrase photosensitive location may refer to an exposed photosensitive portion of a feature of a device without any covering, protective layer, shielding, or any other feature that may separate the light sensitivity from the surrounding environment or sample.

As used herein, "contact microscope" or "contact microscope" refers to any imaging device or technique that includes a photosensitive sensor in contact with a sample to be imaged. For example, a contact microscope may include: (a) a high resolution sensor or set of high resolution light emitting locations with a high optical density exposed to the environment of the device surface along with (b) a device with a portion of the sample to be imaged associated with the surface and, in the case of the light emitting locations, relatively far away from the light emitting locations and the sample's photodetector such that a portion of the sample is in contact (or nearly in contact) with the surface and the sensor can obtain a usable high resolution image when a portion of the sample is in place.

As used herein, "sensor" refers to an integrated circuit, or a component of an integrated circuit that includes a light sensitive element. For example, the sensor may be a component that receives light at the light sensitive element and generates a signal or data indicative of the intensity of the light detected by the light sensitive element, and processes any electronic element that directly drives the light sensitive element or causes the signal or data generated by the light to be conveyed by the light sensitive element.

As described herein, a "parallel" arrangement of surfaces may include a substantially parallel arrangement between the top of the chamber and the surface of the light sensitive sensor, such that the arrangement provides a uniform distribution of particles on the surface of the light sensitive sensor.

As used herein, "sedimentation" refers to placing the surface of a body on a sample such that the body steadily sinks towards the top of the sample. For example, the body may settle on top of the sample if, for example, the body is not attached to or held in place by a separate component. In other cases, the surface of the body may settle on top of the sample based on the body being pressed against the sample.

Overview of the System

In contact microscopes, the sample to be analyzed is associated with the light-sensitive feature of the sensor, since it is, for example, in direct contact (e.g., without any intervening material) with the light-sensitive feature of the sensor or the light imaging of the light source, or nearly in contact with the light-sensitive or light-emitting feature. For example, "nearly touching" may refer to, for example, being within the near field of a light sensitive or emitting feature, which in some cases refers to a distance that is within 1/2 of the wavelength of the light involved or possibly a distance that is within the wavelength range of the light involved.

In embodiments of the systems and techniques we describe herein, one or more devices can be used to correlate a sample with a sensor surface. Such correlation may include any mechanism that facilitates movement, flow, transport, placement, or presentation of, for example, a portion of the sample in contact or near contact with the photosensitive location, including any mechanism that uses mechanical, electrical, electromechanical, pneumatic, hydraulic, capillary action, surface wetting, gravity, and the like.

A. System component

Fig. 1 shows an example of a system 100, the system 100 generally including various components for capturing a high resolution image of a sample 101 in contact with or in close proximity to a surface 103 of a light sensor 102. The system 100 further comprises a light source 119, sample management devices 131 and 133, an integrated chip 104, a headboard 106, a control unit 108, a user device 110, and a user interface 109.

The photosensor 102 includes a two-dimensional arrangement of photosensitive elements 105, which may correspond to an array of pixels in a captured image. For simplicity, the elements of the photosensor 102 are described herein as "pixels". High resolution images may be captured using various color schemes (e.g., full color, grayscale, black and white) or a combination of color schemes. Additionally, sample 101 may be in various phases (e.g., gas, liquid, solid) or a combination of these or other phases.

The light sensor 102 may also include other components as part of, or in addition to, the light sensitive element 105 that perform various functions. For example, the components may drive or read the sensing elements and generate, process, and transmit electronic signals to other components of the system 100 (e.g., headboard 106, control unit 108, user device 110). The components of the light sensor 102 may also perform other functions, such as receiving data transmissions from the components of the system 100.

The sensor 102 may be a component of the integrated circuit chip 104 or formed on the integrated circuit chip 104, which may be fabricated in a uniform fabrication mode, a hybrid fabrication mode, or other conventional fabrication techniques. The chip 104 may be mounted on a headboard 106, which headboard 106 may be part of the control unit 108 or connected to the control unit 108.

The control unit 108 may be part of the user device 110 or connected to the user device 110. User device 110 may provide a user interface 109 for access by a user 115 to adjust and control the operation of system 100. For example, the user device 110 may receive information 111 (e.g., commands) from the user 115 through the user interface 109, process the received information 111, and send the received information 111 to the control unit 108. Further, the control unit 108 may receive data 117 (e.g., sensor data from the light sensor 102) from the headboard 106, process the received data 117, and send the received data 117 to the user device 110 for display on the user interface 109. In some cases, the user interface 109 may be operated by the control unit 108 or the headboard 106 or a combination of various components of the system 100.

The light source 119 may be an external light source external to the system 100 (e.g., room light) that provides ambient light for imaging, or a dedicated light source that provides specific illumination and intensity control of the light provided on the sample 101. For example, the light source 119 may be controlled by the user device 110 or the control unit 110 to adjust the intensity, focus, position, orientation, illumination uniformity, and/or other optical properties of the light provided on the sample 101.

Since the sample 101 is in contact with or in close proximity to the surface 103 of the light sensor 102, no additional optical elements are required to refract, collimate or redirect the light to the light sensor 102 for imaging. For example, light 99 from a portion 107 of the sample adjacent to the pixel (or in the path between the incident light 99 and the pixel) will be mostly (in some cases substantially completely) received by the pixel 105. In this arrangement, the light 99 sensed by the pixel array of the light sensor 102 is directly representative of the corresponding partial array of the sample 101, and thus effectively represents a high resolution image of the sample 101.

The sample transport and management devices 131, 133 may include mechanical or electrical components, or a combination thereof, that facilitate loading and transport of the sample 101 to a location on the surface 103 of the sensor 102 for image capture of the sample on the surface and formation of a thin, uniform layer, such as a monolayer. For example, the devices 131, 133 may be used to move a container including the sample 101 horizontally or vertically along the surface 103 to position the sample 101 in an optical position above the sensor 102 and to hold the container in the optical position during imaging. The devices 131, 133 may also process the sample before and after the imaging process. For example, the devices 131, 133 may be used to mix chemical reagents with the sample 101 during sample preparation, remove chemical reagents from the sample 101 for purification, withdraw the sample 101 from an external source, process the imaged sample after the imaging process, or any other function that may be used with respect to the sample 101 for the imaging process.

User device 110 may be any type of electronic device capable of generating a user interface for receiving and transmitting data communications. For example, the user equipment 110 may be a handheld device, such as a cell phone, tablet computing device, or laptop computing device, or a stationary device, such as a desktop computer or workstation. In some cases, the user device 110 may also be any type of instrument that the user 115 uses to adjust the functions of the control unit 108.

As described in more detail below, the system 100 also includes a chamber top 95 (or "lid," "cover," or "chamber wall" as described herein) that can abut, contact, surround, or enclose the chamber adjacent an exposed surface 103 of a photosensor 102 that holds a portion of the sample 101. A detailed description regarding the operation of the system 100 in relation to the use of the chamber top 95 is provided below. In some embodiments, the chamber top 95 may be configured to be able to be lowered to contact the sample 101 and adjust the volume of the sample 101 (e.g., the volume determined by the area of the sensor and the thickness of the sample layer atop the surface 103 of the light sensor 102). As an example, the adjustment may be made by lowering the bottom surface of the chamber top 95 relative to the sample 101 so that excess sample 101 flows horizontally out along the surface 103 of the light sensor 102. The chamber top 95 may also be lowered in other ways, as described in more detail below. As described herein, the space formed between the bottom surface of the chamber top 95 and the surface 103 of the photosensor 102 forms a "chamber" for the sample 101 once the descent of the chamber top 95 is complete. Thus, the volume of the sample 101 initially placed on top of the surface 103 is greater than the volume of the sample 101 within the chamber because excess volume of the sample 101 (e.g., the difference between the volume of the sample 101 introduced and the volume of the chamber) is removed from the chamber as a portion of the sample 101 flows out of the chamber after the chamber top 95 is initially contacted with the sample 101 and before the chamber top 95 reaches its final placement. In some cases, excess volume of sample 101 flows laterally out to surface 103. In other cases, the bottom surface of the chamber top 95 may be a porous surface, which allows excess sample 101 to flow out of the chamber through the pores of the chamber top 95. In these cases, the size of the pores may be such that only fluid passes through the pores, but the particulate matter of sample 101 is too large to pass through the pores.

Although fig. 1 shows various components of the system 100, a commercial product associated with the system 100 need not include each of the components depicted in fig. 1 and described herein (and may include components other than those shown). In various embodiments, any combination of two or more of the light sensor 102, the chip 104, the headboard 106, the control unit 108, and the user device 110 may have various mechanical and electrical connections therebetween. In addition, the mechanical, fluid flow, electronic, software, data processing, communication, storage, and electrical functions required for various operations may be distributed in various ways among the various pairs and among three or more of those portions of the system. The distribution of functions may be arbitrary or may be based on business and technical considerations in various ways.

B. System operation

During operation, the photosensor 102 detects incident electromagnetic radiation 99 (or "light") generated from the light source 119 and scattered from or emitted from the sample 101. The wavelength of light passing through, scattered from, or emitted from the sample 101 may change, for example, as it passes through or is scattered or emitted. The incident light 99 and the transmitted, scattered, or emitted radiation are typically in the visible, near ultraviolet, or near infrared wavelength ranges. However, as described herein, light 99 may include light from all of these ranges.

To capture an image of the sample, the light sensor 102 is driven and read during an image capture period. During an image capture cycle, light 99 received by the photosensor 102 at each of its pixels is converted to an electrical signal (e.g., an analog signal or a digital value) that is passed to the electronic components of the chip 104. Signals may be read in parallel or in series, depending on the components of chip 104. The electrical signal from each pixel is typically represented by a quantized intensity value corresponding to the light intensity sensed by the pixel, within some range, such as a range represented by, for example, a 16-bit digital value.

The color information may be obtained in various ways, for example using bandpass optical filters on a plurality of adjacent pixels, or sequential imaging with different color illumination, etc. The electrical signals received from the various spatial pixels may represent a full color high resolution high dynamic range image of the specimen 101. In addition to the electronic features of system 100, there are also mechanical elements discussed below that process, contain, and illuminate sample 101.

Sample preparation

A. Characterization of the sample

Sample 101 (also interchangeably referred to as a "sample") may be any type of phase (e.g., liquid, solid, gas) or combination in direct contact with surface 103 of light sensor 102. In some cases, sample 101 is a fluid that includes various types of particulate matter, such as cells (e.g., human or animal blood cells, mammalian cells, bacterial cells, and/or plant cells), molecules (e.g., DNA, RNA, peptides), proteins (e.g., antigens and antibodies), or contaminants in environmental or industrial samples. In this case, sample 101 may be dispensed into a chamber above surface 103 and operated using devices 131, 133 to position sample 101 on light sensor 102.

Referring to fig. 2, a sample 101 being imaged may include or consist of small, similar types of cells 97, such as particles, bits, spots, cells, or molecules, or combinations thereof or combinations of any two or more different types. The cells 97 may be suspended in a liquid 101 or entrained in a liquid 101 to form liquid suspended particles 97, entrained in a gas to form gas suspended particles 97 (not shown), resting in an unsuspended and unentrained form (e.g., a powder) on a surface 103 of a photosensor 102 (not shown), or held in an integrated matrix of solid, gelled, or other integral self-supporting material such as a segmented layer of tissue or the like. As described herein, a "matrix" may include, for example, any material in which particles 97 are retained, including liquids, gases, solids, gels, or other materials.

Sample delivery

A. Distribution technique

For some embodiments of the systems and techniques described herein, fig. 4A illustrates a top view of the system 100 during a sample dispensing process. As shown, a predetermined volume of sample 101 is dispensed onto the surface 103 of the light sensor 102 before the imaging process is performed. Using the flow director 1050, a volume of the sample 101 is dispensed by using the fluid loading pipette 1040 to bring the pipette tip 1052 close to a predetermined position such that the sample 101 is deposited on top of the surface 103.

As described in more detail below, various types of dispensing techniques may be used to deliver a volume of sample 101 onto surface 103. In some cases, fluid loading pipette 1040 is a particular type of pipette, referred to herein as a "duplex pipette. In some cases, fluid loading pipette 140 is a conventional micropipette.

In some embodiments, the chamber top 95 and/or surface 103 of the sensor 102 is coated with a hydrophilic coating to enhance capillary forces and increase the speed of the sample transport process. In some embodiments, a hydrophobic coating may be used around the sensor active area to contain the liquid sample. In cases where settling of particles 97 is an important issue, sample 101 may be mixed, for example during fluid ejection and/or lowering of chamber top 95, one or both of which may be automatically controlled, through the use of pumps, actuators, and other techniques.

B. Double pipette

Fig. 4B and 4C show schematic diagrams of an example of a fluid loading pipette 1040a, referred to herein as a duplex pipette 1040 a. Referring to fig. 4B, a double pipette 1040a includes two volume capillaries 1042a and 1042B that deliver separate input fluid streams (e.g., a blood sample and a diluent/chemical stain) to a mixing well chamber 1044 that combines the two input fluid streams into the mixing well chamber 1044 with an aperture at the other end for dispensing the mixed fluid of the two input fluid streams.

Fig. 4C shows the internal structure of the hybrid well chamber 1044. As shown, the left side portion of the mixing well chamber 1044 includes two receiving ports with the ends of the volume capillaries 1042a-b attached to the mixing well chamber 1044. In some embodiments, the mixing well chambers may be detachable from the fluid containers 1042a-b such that a single mixing well chamber 1044 may be reused for multiple deliveries of a single sample 101. The two receiving ports converge into a single channel, including a recess 1046 to help combine the two input fluid streams into a single output. For example, the grooves 1046 may be arranged transverse to the fluid flow through the fluid channel such that the grooves 1046 interfere with the fluid flow and enhance the combination of the two input fluid streams, as previously described in the scientific literature (Sabotin i., tsto g., bissaco g., Junkar m., and Valentincic J. (n.d.); interleaved chevron mixers designed for EDM machining;retrieval 7 months and 7 days in 2015; fromhttp://lab.fs.uni-lj.si/lat/uploads/ edm/bibJoze/10-imbt.pdf) The method as described in (1).

C. Spacing feature

A wide variety of techniques and devices may be used to form and maintain the height (e.g., precise height) of gap 220. As described herein, such techniques are commonly referred to as "spacing features". In the example shown in fig. 2, the spacing features comprise microspheres or other kinds of uniformly sized beads. By way of example, in some embodiments, the spacing features 230 are monodisperse, rigid polymeric microspheres having a precisely defined diameter (e.g., 3.00 μm, with a coefficient of variation of less than 5%). In this example, to establish accurate and uniform spacing of the gap 220, which correlates to the volume of the sample 101 between the chamber top 95 and the surface 103, precision in bead size can be used to ensure that the gap 220 is repeatable over multiple imaging processes.

In some cases, for a given kind of sample unit or accurately specified volume of sample (e.g., for blood counting, or other analysis, where the number of particles 97 is counted for an accurate volume of sample), the volume of sample 101 to be imaged is accurately controlled by the width and length of the top surface of light sensor 102 and the height of the gap 220 (or chamber) between surface 102 and the flat bottom surface of chamber top 95. In some cases, the volume may not need to be precise, but the gap height may need to be a precise amount, or no greater than a certain amount, or no less than a certain amount, or a combination of these conditions.

As shown in fig. 2, in some embodiments, the spacing features 230 are included within a sample, such as a sample with a liquid matrix, in which particles 97 (which may be smaller than beads) are suspended when the sample is delivered to the sensor surface 103. A uniform and precise gap height can be achieved if the top of the chamber is allowed to settle or press on the sample and provided that there are sufficient beads in the sample and that they are fairly evenly distributed in the liquid. To this end, for example, the beads may be present in the sample at a rate of 10000-500000 beads per microliter of sample. If the beads are selected to have near neutral buoyancy in the sample, a uniform distribution of the beads in the sample can be maintained by simple mechanical agitation.

In some cases, the beads may be about the same size as the particles 97. In some embodiments, two different sizes of beads may be included. The larger dimension defines the desired spacing. The smaller size can be calculated to verify that the volume of the sample space is as expected, assuming that the smaller beads are reasonably evenly distributed in the sample and that the number of smaller beads per unit volume of sample is known. The beads may be transparent to allow light to pass through the sensor, or may be colored, or fluorescent, or opaque, or a combination of two or more of these features.

In some embodiments, instead of using the spacing features 230 included within the sample 101, the height of the chamber (e.g., gap 220) formed instead between the bottom surface of the chamber top 95 and the surface 103 may be maintained by an array of posts that protrude from the surrounding surface around the surface 103 (e.g., on the surface of the headboard 106). In such embodiments, the headboard 106 of the receiving surface 103 can be specifically manufactured so that the posts have a predetermined height corresponding to the optical gap 220 required for a particular imaging procedure. In operation, after the sample 101 is introduced, the chamber top 95 can then be lowered onto the surface 103 until the bottom surface of the chamber top 95 contacts the top surface of the pillars. Various aspects of the pillar array (e.g., array pattern, pillar density) may also be adjusted to affect the distribution of particles 97 along surface 103.

In some cases, the amount of sample 101 loaded onto the light sensor 102 is greater than the amount required for imaging. In some embodiments, the sample 101 needs to be in the form of a relatively thin layer (e.g., 1 μm to 100 μ), or have a thickness such that a monolayer of cells of the sample are displaced on the sensor for imaging. In this case, the chamber top 95 may be lowered to contact the sample 101 and adjust the volume of the sample 101 (e.g., the thickness of the sample layer atop the surface 103 of the photosensor 102).

D. Sedimentation after distribution

As described herein, it may be desirable for the concentration of the sample 101 to be imaged to be the same as or have a predetermined relationship with the volumetric concentration of the sample initially dispensed on the surface 103. In some cases, the weight of particulate matter (e.g., particles 97 and spacing features 230) within sample 101 is heavier than other fluid components of the sample (e.g., diluent), which makes the particulate matter prone to build up rather than flowing or moving when a force is applied to the volume of sample 101.

One example of an external force may be gravity, which may cause a settling concentration gradient in sample 101 as particles 97 descend toward the bottom of sample 101 due to gravity. Another example of a force may be a force exerted by the bottom surface of the chamber top 95 during the descent of the chamber top 95, as described herein. In this example, the chamber top 95 accelerates the sample 101 downward beyond the perimeter of the sensor 102, and the heavier suspended particles 97 have more momentum than the fluid components and may not move or accelerate as quickly as other portions of the sample. In this case, particles 97 may remain on the surface 103 of the light sensor 102, resulting in a higher concentration than the volume concentration in the sample 101 dispensed on the surface 103 before removing the excess volume of the sample 101. In yet another example, the force may also include a friction force between sample 101 and various surfaces of the system (e.g., surface 103, surface 1006, etc.) or a shear force generated within the sample due to interaction with such surfaces. Frictional and shear forces can reduce the velocity of the particles 97 relative to the sample flow.

In addition, after the top of the chamber has finished its descent, the sample may continue to flow, causing the particles 97 to move and disrupt their imaging. In some embodiments, the viscosity of the sample can be adjusted to control the concentration of particles 97 and reduce the flow of the sample during imaging. In some examples, the adjustment may be accomplished by adding one or more viscosity control agents to the sample. The settling rate of the particles 97 can be reduced and the fluid can be allowed to exert a stronger force on the spacer beads and particles 97 to counteract their momentum and friction. The increased viscosity may also reduce the likelihood of flow after the top of the chamber has completed its descent. Suitable agents may include dextran, glycerol, starch, cellulose derivatives such as methyl cellulose, any combination of these materials, and other materials.

Alternatively or additionally, one or more reagents may be added to the sample to increase the diluent density, thereby reducing or even eliminating the density difference between the diluent and the spacer beads and/or particles 97. The reduced or eliminated density difference may also control the concentration of particles 97 and reduce the flow of sample during imaging.

The agent for increasing the density of the diluent may be the same agent as the viscosity controlling agent. In some embodiments, thixotropic agents may be used to achieve the same effect, and also allow easier mixing of the particles 97 with the diluent. In some cases, photo-crosslinkable reagents or gelling agents (e.g., temperature-dependent, such as low melting point agarose) can be used to increase the sample viscosity while allowing easy mixing of the particles 97 and diluent. For example, a sample with suspended particles 97 and a gelling agent such as liquid agarose may be initially squeezed by the chamber top 95 to form a monolayer of particles 97 on the surface 103. The temperature of the sample can be cooled to form an agarose gel structure that "traps" the particles 97 in their place within the monolayer, which can then be used, for example, to perform comet-testing for DNA damage. For example, to perform a comet assay, the sample may include a DNA intercalating stain for detecting particles 97 that may be cancer cells. In this case, after gelation, the top of the chamber may be raised briefly to allow the cell lysis medium to penetrate the gel; a voltage gradient may then be created along the length or width of the chamber by electrodes that may also be placed on opposite ends of the chamber (e.g., on two opposite sides of the chamber top 95 that extend to opposite edges of the truncated surface 1102 of the chamber top). In other cases, polyacrylamide, starch, or other gels may be used to achieve rapid and inexpensive electrophoretic analysis of proteins, nucleic acid side-insertion, and other macromolecules. The electric field generated by the electrodes can be used to induce movement of small particles suspended (e.g., not captured within the gel), and this movement can be monitored using the image sensor 102 to measure the surface charge or zeta (zeta) potential of the particles 97.

Chamber top descent

E. Reduction technique

Once sample 101 has been dispensed on surface 103 of light sensor 102, chamber top 95 may be lowered toward surface 103 to remove excess volume of sample 101 atop surface 103, thereby creating a thin layer of particles 97 (e.g., cells dispensed in a fluid sample) to be uniformly distributed on surface 103. In some embodiments, the removal of excess volume is performed in such a way that removal of excess volume does not change the volume concentration of particles 97 above the surface 103 of the photosensor 102, such that the relatively small volume of sample 101 imaged (e.g., about 40nL) represents a large volume of sample (e.g., about 50 μ L or more) dispensed onto the surface 103 of the photosensor 102. In other embodiments, the removal process produces a new concentration of particles 97 within a relatively small volume of the sample 101 that is consistently proportional to the volume concentration of particles 97. In such embodiments, a correction factor may be determined and applied to the captured data to derive a desired sample concentration for imaging. For example, to obtain a desired sample concentration for imaging, sample 101 may be further processed using techniques described further below.

The chamber top 95 may be lowered in various ways, as described in particular below for various embodiments. In the example shown in fig. 2, the chamber top 95 has a flat bottom surface 200 that is lowered towards the surface 103 such that the surface 200 remains substantially parallel to the top surface 103 of the sensor 102. As described herein, this type of droop is referred to as a "linear droop". Figure 3A shows another example where the chamber top 95 is initially positioned in an inclined position such that a first edge of the chamber top 95 contacts the surface 103 along a line of contact and an opposite edge of the chamber top 95 is away from the surface 10. In this configuration, the opposing edges of the chamber top 95 then descend along the axis of rotation defined by the line of contact between the first edge of the chamber top 95 and the surface 103. The chamber top 95 may be lowered with a controlled velocity profile until a point 1006 on the bottom surface of the chamber 95 is flush with the surface 103. As described herein, this type of descent is referred to as a "pivoting descent".

In some cases, data such as position variables or parameters that control the lowering of the chamber top 95 may be selectively selected based on the type of sample 101 used and then stored for later use. The stored data may then be accessed and automatically applied to the configuration of the system 100 using, for example, a controller. The degradation can then be performed with sufficient repeatability for different imaging procedures based on the stored data.

In addition, various mechanisms may be used to control the lowering of the chamber top 95. For example, the chamber top 95 may be lowered manually by a person using physical means (e.g., a circular knob) or automatically using a machine such as an actuator 1010.

In some embodiments, after an initial descent of a first edge of the chamber top 95 facing away from the surface 103, a corresponding point on the bottom surface of the chamber top 95 contacts the sample 101 throughout the descent, while the opposite end of the chamber top 95 may be repeatedly raised and lowered (e.g., not all the way down to a final position). This repeated movement of the chamber top 95 may cause the sample 101 to flow into and out of the space formed between the surface 103 and the chamber top 95, which may be used to create a mixing effect on the sample 101 to evenly distribute the particles 97 along the surface 103 prior to the imaging process.

In some embodiments, the chamber top 95 has a surface 1004 that presses against a surface 1005 of the holder 1012, which facilitates the lowering of the chamber top 95. Surface 1005 may be formed of an encapsulating epoxy deposited on surface 103 to form retainer 1012. The linear point of contact between surface 1004 and surface 1005 may then be used as a hinge for lowering or raising the chamber top 95.

As an example of use, after depositing the sample onto the surface 103 of the light sensor 102, the chamber top 95 is held at an angle by another contact point 1006 elsewhere and slides forward until the surface 1004 is pushed towards the surface 105 so that it cannot slide further. The hinge then allows the chamber top 95 to be rotationally twisted along its axis of rotation such that the edge of the chamber top 95 opposite surface 1004 is lowered toward surface 103. The chamber top 95 is then slid along the surface 1005 until the adjacent edge of the chamber top 95 strikes another barrier 1007 (e.g., also a separate structure that is part of or a side of the package). This allows the positioning of the chamber top in the y-direction to be repeated from test to test (or sample to sample). The contact point 1006 holding the top of the chamber is then lowered, allowing the top of the chamber to hinge down until flush with the sensor. In some embodiments, the contact point is lowered such that its friction with the top of the chamber provides a small force that pushes the top of the chamber toward the ridge rather than pulling it away to reduce or avoid interference with the position of the top of the chamber at the wall 1005. It is possible that the top of the chamber may slide after placement on (or down to) the sensor and as the sample is expelled from the chamber. Guide posts 1008 and/or walls near the sides of the sensor are sometimes used to minimize the distance the chamber top can travel.

In some embodiments, the contact edge 1004 at the top of the chamber has two extended points at opposite ends 1009 to allow sample flow between the points in the hinge direction. This can increase the uniformity of the sample flow in all directions from below the top of the descending chamber, reducing the artificial non-uniform distribution of particles 97 (e.g., cells).

In some cases, the actuator 1010 may be a passive device that is not secured to the chamber top 95 and is used to lower the chamber top 95. The chamber top 95 may rest on the actuator 1010 and descend by gravity or another force (e.g., magnetic, electromagnetic, spring). The descending velocity profile may be controlled by various means including, for example, a rotating weight, a buffer cylinder 1011, a magnet, an electromagnet, and the like.

Although the chamber top 95 is described as descending toward the sensor surface, the described mechanism may be used with any surface (e.g., a slide) in an embodiment (e.g., counting cells or other particles using a standard microscope).

Blood analysis

One particular set of applications for the system 100 relates to the analysis of blood samples. In these applications, the system 100 may be used to detect and analyze cell types (e.g., white blood cells, red blood cells) in blood. The system 100 can be used to count various types of cells, determine normality of blood cells, monitor blood cell function, and analyze blood chemistry.

A. Leukocyte concentration and count calculations

The concentration of White Blood Cells (WBCs) in the blood is relatively low and can be further reduced by any diluent added to the blood at the time of sample preparation. As a result, the total number of leukocytes on the sensor surface to be imaged or counted can be low. Typically, the counting error of a particle is the square root of the count, and a low number of particles to be counted may result in a high percentage error.

In some embodiments, the leukocyte concentration can be increased in a predictable manner. In some embodiments, suitable spacer beads may be used so that the average concentration of Red Blood Cells (RBCs) may be maintained at a desired level on the sensor surface while the blood count is increased. In general, as the chamber top 95 is lowered toward the sample, cells that are in contact with the surface of the chamber top 95 and the surface 103 of the sensor 102 in opposite directions may be captured. For example, cells generally do not move when they are compressed between opposing surfaces. Thus, the size of the spacer beads can be chosen such that the distance between the top of the chamber and the surface of the sensor is smaller than the average diameter of the leukocytes. In some cases, to maintain the concentration of red blood cells, the diameter of the beads may be greater than the average diameter of the red blood cells. The descending chamber top compresses white blood cells having a diameter greater than the diameter of the beads, while not compressing red blood cells having an average thickness less than the diameter of the beads. As the top of the chamber descends to the bead diameter and the total volume of sample decreases, the concentration of leukocytes on the sensor surface increases. An example of a bead diameter may be 5 microns. Other suitable diameters may be selected to control the concentration of different cell types in the sample.

Once the chamber top 95 has been lowered to its final height, the height of the chamber (e.g., height 220 shown in fig. 2) and the surface area of the surface 103 of the sensor 102 may be used to calculate the volume of blood imaged on the surface 103. The white blood cell concentration may be increased in proportion to the cell size relative to the concentration of smaller uncaptured cells, such as red blood cells. The relationship between the size and concentration of leukocytes is integrated over all leukocyte sizes to obtain an average concentration (e.g., the volume concentration in the sample before cell concentration). This concentration effect can lead to useful improvements in count statistics.

Various products can be manufactured and delivered according to the architectures and principles we discuss. The product may include a sensor unit, a sensor unit + reading unit, a sensor unit plus + headboard, a sample chamber, a chamber top (or lid), a sensor unit + pipette, a sensor unit + pump, system equipment, hand-held devices, inserts and accessories for other devices, a pipette, a pre-loaded pipette, an image processor, software, a light source, a sample chamber + light source + sensor + headboard + electronics in a complete device, and combinations of two or more of these and other components.

In considering the wide range of operations performed by sensors and systems, as well as a wide range of applications, it may be useful to recognize that some involve imaging, some involve analysis, and some involve a combination of analysis and imaging.

Examples of the invention

The following examples of embodiments of the system 100 use various techniques for sample loading and processing, and/or for lowering the chamber top onto the sensor surface prior to performing the imaging process. As described in more detail below, each embodiment provides advantages that may improve aspects of the imaging process.

Example 1 open Chamber apparatus

Fig. 5A-5B illustrate perspective views of an open chamber device 1100 that may be used to perform full blood counts, as described in the present disclosure, as well as other types of tests (e.g., bio-dosimetry). In this embodiment, the chamber top 95 is lowered onto the surface 103 of the photosensor 102 using a carrier arm that is lowered using an actuating element. The chamber top 95 is initially placed on the extended tip of the carrier arm such that the chamber top 95 is not rigidly attached to the carrier arm, but is loosely attached to enable the chamber top 95 to settle on top of the extended tip of the carrier arm. In addition, the chamber top 95 is placed in such a way that its descending direction is substantially parallel to the surface 103 of the light sensor 102.

Referring now to fig. 5A and 5B, a system 1100 includes a plate 1110 having an open sample chamber 1160. The surface of the plate 1110 includes the surface 103 of the light sensor 102 and the headboard 104. A more descriptive view of open sample chamber 1160 is shown in fig. 6C. The system 1100 also includes a carrier arm 1120 and a support structure 1140 attached to the actuating device 1130 for positioning the chamber top 95 in a substantially parallel manner over the surface 103 prior to operation.

In operation, the chamber top 95 is initially placed over the surface 103 onto an extension of the carrier arm 120 (shown in fig. 6E as an extended tip 1124). After placement on the extended tips 1124 of the carrier arms 1120, the chamber top 95 is also positioned parallel to the surface 103 by inserting guide rods attached to the chamber top 95 (shown in fig. 6B as guide rods 1104a and 1104B) into holes on the support structure 1140. Inserting the guide rods 1104a, 1104b into the holes of the support structure 1140 ensures that when the carrier arm 1020 is lowered, the corresponding lowering of the chamber top 95 results in a "linear lowering", as described above. A more detailed description of each of the individual components of the system 1100 is provided below.

B. Top of the chamber

Fig. 6A and 6B show perspective and top views, respectively, of a chamber top 95 for use with the open chamber device 1100. As shown, the chamber top 95 includes a set of guide rods 1104a and 1104b that serve to initially position the chamber top 95 on the extended tips 1124 of the carrier arms 1120 and also ensure that the initial position of the chamber top 95 is substantially parallel to the surface 103.

The chamber top 95 additionally includes a membrane 1104 (shown in fig. 6D) comprising truncated pyramidal members 1102 extending from a bottom surface 1106 (shown in fig. 6A) of the chamber top 95. In operation, as the chamber top 95 is lowered using the carrier arm 1230, the top surface of the truncated pyramid 1102 faces the surface 103 as the chamber top 95.

In some cases, film 1104 is a flexible film that is spread over a rigid frame. The membrane is "elastic" in the sense that it is capable of deforming when a force is applied to its surface and then of conforming back to a flat surface after the applied force is removed. For example, when the chamber top 95 is lowered, a flexible membrane may be used to prevent rigid forces from being exerted on the sample top above the surface 103. This ensures that the top of the truncated pyramid 1102 pushes the sample downwards due to only a gentle predetermined force, so that excess volume is displaced from the chamber formed between the truncated top of the pyramid 1102 (i.e. the surface facing the surface 103 of the light sensor 102) and the surface 103 of the light sensor 102.

The top surface of truncated pyramidal member 1102 may be designed such that its surface area corresponds to surface area 103. Furthermore, truncated pyramid member 1102 is constructed of a transparent material (e.g., glass, plastic, acrylic, etc.) such that light 99 generated by light source 119 can pass through truncated pyramid member 1102 and reach light sensor 102 to collect an image of the volume of sample 101 placed between the top surface of truncated pyramid member 1102 and surface 103 of light sensor 102.

Although the truncated pyramid member 1102 described herein is constructed of a transparent material (e.g., glass or plastic) to allow light to be transmitted into the sample and then to the light sensor 102, in some embodiments, the truncated pyramid member 1102 may be constructed of an opaque material for use in dark field illumination microscopy, where light scattered by the sample is only detected on the light sensor 102. In other embodiments, the top surface of truncated pyramidal member 1102 may also be modified to be transparent only to a limited wavelength of light, using a pigment of a particular color within the transparent material of the member or on its top or bottom surface, or by depositing a thin film spectral filter on the top or bottom surface.

The chamber top 95 may additionally include a set of weighting elements 1108 that evenly distribute the weights along the bottom surface of the chamber top 95 such that the chamber top 95 descends substantially parallel toward the surface 103 as the carrier arms 1120 are lowered. Although fig. 8 depicts an example of an arrangement of weighted elements 1108, in other embodiments, weighted elements 1108 may be positioned in other arrangements as long as the arrangement provides a means of lowering chamber top 95 substantially parallel to surface 103.

C. Open sample chamber

Fig. 6C shows an example of a top view of open sample chamber 1160. Open sample chamber 1160 includes a surface and chip 104 as previously described with respect to fig. 1.

D. Carrier arm

Fig. 6E shows an example of a top view of the carrier arm 1120. As described herein, the carrier arm 1120 includes an extended tip 1124 that freely supports the chamber top 95 in its initial placement. In operation, the actuation device 1130 of the system 1100 is used to manually or automatically lower the height of the carrier arm 1020 relative to the surface 1132 of the base of the system 1100 (as shown in fig. 8) such that as the height decreases, the chamber top 95 is lowered toward the surface 103 of the photosensor 102.

The carrier arm 1120 is able to descend to a height relative to the open specimen chamber 1160 such that after a height, for example at the height of the open specimen chamber 1160 from the base of the system 1100, the extended tip 1124 of the carrier arm 1120 no longer supports the chamber top 95 due to the top surface of the truncated pyramidal member 1102 being in contact with the sample 101 placed on top of the surface 103.

Once the height of the carrier arm 1120 from the surface 1132 of the base is less than the height of the sample chamber 1160, the chamber top 95 is free to settle on top of the surface 103 rather than on the carrier arm 1120, which causes an excess volume of sample 101 placed on top of the surface 103 to flow out of the chamber formed by the top surface of the truncated pyramidal member 1102 and the surface 103, as previously described with reference to figure 2. In this regard, gravity exerted on the chamber 105 may be used to form a substantially uniformly distributed volume of the sample 101 on the surface 103 without the use of external forces as described herein with respect to other embodiments.

EXAMPLE 2 Point of Care device

Fig. 7A-7C show perspective views of a point-of-care blood counting apparatus 1200 that can be used in resource-limited areas and/or other areas without access to conventional laboratory bench-top reagents and equipment. In this embodiment, the contact microscope system is housed within a portable housing 1210 that includes a compartment for the mobile device 1220 and a compartment for the portable microscope arrangement, as described in more detail below. In some cases, the portable microscope device may comprise a more complex device as shown in fig. 8, including a latching mechanism and a rotational damper to lower the chamber top 103.

In general, device 1200 is capable of capturing an image of a blood sample without the need for any external equipment other than the sample distribution device shown in FIGS. 4B-4C. The user device 1220 may be any type of mobile computing device capable of performing computing operations and capturing images. In some embodiments, the user device 1220 includes software (e.g., a mobile application) that enables a user to capture images of blood samples without extensive training or sample preparation.

In some cases, device 1200 may be used in resource-limited areas of developing countries where the operator performing the blood count test lacks the training required to perform blood counts using traditional microscopy techniques. In this case, the device 1200 may be used to provide a low-easy-to-use portable device to accurately provide blood cell counts with limited sample preparation and processing. For example, the user device 1220 may provide an interface that enables an operator to dispense a volume of sample 110 into a portable microscope device and then capture an image of the dispensed blood by providing simple user input on the user device 1220. Specific descriptions relating to the components of the portable microscope device are provided in more detail below.

A. Portable microscope device

Fig. 7A-7C show various views of the device 1200, including a compartment for housing a portable microscope device. The apparatus includes a carrier arm 1230, a slot 1232 for holding the top of the chamber, a headboard 1250 with a sample recess 1260, and a sample delivery module 1240 with a pipette well 1242. The chamber top 95 may be attached and/or configured to the carrier arm 1230 in various configurations. In some cases, the chamber top 95 is a separable component that includes a truncated pyramid 1102 shown in fig. 6D. In addition, the device 1200 also includes a light source located directly above the carrier arm 1230 (and chamber top 95) when the cover of the housing 1210 is in the closed position, and a sensor (not shown), such as the light sensor 102, located at the bottom of the sample recess 1260.

In operation, the initial configuration of the carrier arm 1230 is facing up to enable an operator to prepare the device 1200 for an imaging operation, as shown in fig. 7B. The chamber top 95 is inserted into a slot 1232 in the carrier arm 1230 such that when the carrier arm is fully lowered, the top surface 1102 of the truncated pyramid will face the surface 103 within the sample recess 1240. A pipette may then be used and the tip of the pipette inserted through the aperture 1242 of the sample delivery module 1240 to introduce a volume of the sample into the sample recess 1260, as shown in fig. 7C. The aperture 1242 may be sized for use with the particular type of pipette being used and for dispensing a volume of sample into the sample recess 1260. For example, for larger sized pipettes, the bore 1242 may be larger such that the tip of the pipette dispensing the sample volume is above the center point of the sample recess 1260 when the corresponding pipette is inserted into the sample bore 1242. In some cases, sample delivery module 1240 may be interchangeable such that a single device 1200 may be used with different types of pipettes.

The carrier arm 1230 is then lowered toward the headboard 1250 once the volume of the sample is dispensed into the sample recess 1260. For example, when the carrier arm 1230 is lowered with the chamber top 95 toward the headboard 1250, stopping at a position set by the thickness of the slot feature 1232, with the chamber top 95 resting on the spacing feature 230, no longer supported by the lower flange of the slot 1232. In this configuration, after the carrier arm 1230 is lowered to its final position, as described above, the top surface of the truncated pyramid can then be pressed against the sample volume dispensed in the sample recess 1260, causing excess sample volume to flow out of the chamber defined by the top surface of the truncated pyramid 1102 and the surface 103, as described previously with respect to the open chamber device. Once the carrier arm and chamber top 95 are in this position, the lid of the housing 1210 can be closed to exclude extraneous light and an image of the sample can be captured using the user device 1220 as a controller for the light sensor 102 under the sample recess 1260.

B. Improved closing mechanism device

In some embodiments, the portable microscope device of the device 1200 includes the improved closure mechanism device 1300 shown in fig. 8. The device 1300 is similar to that of the device 1200 shown in fig. 7A-7C, but includes additional mechanical components, such as a latch mechanism 1320, a spring (not shown) that drives the carrier arm down once the latch is released, and a rotational damper 1340 for adjusting the rate of carrier arm down, to more effectively lower the carrier arm 1330 onto the surface 103, accurately placing the truncated pyramid of the chamber top 95 on the surface 103 of the photosensor 102 within the sample hole 1240. In this regard, the apparatus 1300 can be implemented into the apparatus 1200 to improve ease of use (e.g., reduce the need to manually lower the carrier arm 1230 in a particular manner) and reduce variation in results between subsequent imaging processes.

EXAMPLE 3 closed Chamber device

Fig. 9A-9D illustrate different views of a closed chamber device 1400 for performing full blood counts, as described throughout this disclosure, as well as other types of tests. In comparison to the previously described open chamber device 1100, the closed chamber device 1400 reduces the need to manually load a sample onto the surface 103 or remove a sample from the surface 103 and enclose the sample so that the operator is not exposed to potentially harmful components of the sample. Additionally, the device 1400 can automatically clean the surface 103 by injecting a volume of cleaning agent into the enclosed chamber.

C. System component

The enclosed chamber device 1440 includes a headboard 1410 attached to an enclosure body 1420 with a set of rigid walls 1430 permanently bonded to the headboard 410 and the enclosure body 1420. The closure body 1420 may be any type of suitably transparent rigid material, such as glass, acrylic, plastic, etc., that enables light from a light source above the closure body 1420 to be transmitted in an enclosed space within the rigid sidewall 1430, as will be described in more detail below. The headboard 1410 may be an integrated circuit board that includes a light sensitive sensor, such as light sensor 102, that has a surface 103 that is exposed to the sample fluid during imaging operations.

Fig. 9B and 9C show a top view and a cross-sectional view, respectively. Once the closure body headboard 1410, rigid side wall 1430, and closure body 1420 are permanently joined together, an enclosed space is formed within the rigid side wall 1430. The exterior of the enclosed space within the rigid side wall 1430 includes a pressure chamber 1432 wherein positive or negative pressure can be applied through holes 1442 in the rigid side wall 1430. The holes 1442 may be placed on any rigid wall as long as the application of negative and positive pressure may be evenly distributed throughout the volume of the pressure chamber 1432.

The pressure chamber 1432 surrounds a set of deformable sidewalls 1440 that surround the fluid chamber 1450. The deformable sidewall 1440 may be made of any suitable solid material that can withstand the pressure applied within the pressure chamber 1432. In some cases, deformable sidewall 1440 may be made of a solid elastomer that is capable of deforming due to pressure applied to pressure chamber 1432. The chamber top 95 of the fluid chamber 1450 is a transparent solid or rigid material that allows light from the light source to pass into the fluid chamber 1450. The chamber top 95 is rigid so that any pressure applied to the pressure chamber 1432 does not deform it, thereby maintaining a smooth flat surface of its surface 103 facing the light sensor 102. The chamber top 95 is secured to a deformable sidewall 1440 to allow the height of the fluid chamber 1450 to be varied due to negative or positive pressure applied to the pressure chamber 1432, as described in more detail below.

D. Operation of

Fig. 9D shows an example of operating the closed chamber device 1440 prior to performing an imaging procedure. As previously described, a sample fluid to be analyzed enters the fluid chamber 1450 through the inlet port 1404 and exits the fluid chamber 1450 through the outlet port 1406. In an initial state, the height of the fluid chamber 1450 is increased to enable injection of the sample fluid into the fluid chamber 1450 (e.g. shown on the left side of fig. 9D). This is accomplished by applying negative pressure to pressure chamber 1432 to create a pressure differential between pressure chamber 1432 and fluid chamber 1450. In some cases, the negative pressure may be provided by applying suction through the aperture 1442 to draw a volume of liquid or gas contained within the pressure chamber 1432. The pressure differential between the pressure chamber 1432 and the fluid chamber 1450 causes the deformable sidewall 1440 to deform to a state 1440a to accommodate the volume of sample fluid entering the fluid chamber 1450, resulting in an increase in the height of the fluid chamber 1450.

Once the appropriate sample volume is delivered in the fluid chamber 1450, the chamber top 95 may be lowered towards the surface 103 to produce a cell monolayer of particles as described in this specification. This may be accomplished by applying a positive pressure to the pressure chamber 1432 such that the positive pressure deforms the deformable sidewall 1440 to the state 1440b to accommodate the increased pressure within the pressure chamber. In some cases, positive pressure may be provided by applying a volume of gas or transparent liquid into the pressure chamber to displace the deformable sidewall 1440 and thus lower the chamber top 95 toward the surface 103. As the height of the fluid chamber 1450 decreases, the excess volume of sample fluid within the fluid chamber exits the fluid chamber 1450 through the outlet port 1406. After the chamber top 95 reaches its final height, an image of the remaining fluid sample within the fluid chamber 1450 may be captured, for example (set by the spacing features 230 previously described with respect to fig. 2).

Other embodiments

A number of embodiments have been described. Nevertheless, it will be understood that various modifications may be made without departing from the spirit and scope of the invention. In addition, the logic flows depicted in the figures do not require the particular order shown, or sequential order, to achieve desirable results. In addition, other steps may be provided, or steps may be eliminated, from the described flows, and other components may be added to, or removed from, the described systems. Accordingly, other embodiments are within the scope of the following claims.

Various products can be manufactured and delivered according to the architectures and principles we discuss. The product may include a sensor unit, a sensor unit + reading unit, a sensor unit plus + headboard, a sample chamber, a chamber top (or lid), a sensor unit + pipette, a sensor unit + pump, system equipment, hand-held devices, inserts and accessories for other devices, a pipette, a pre-loaded pipette, an image processor, software, a light source, a sample chamber + light source + sensor + headboard + electronics in a complete device, and combinations of two or more of these and other components.

In considering the wide range of operations performed by sensors and systems, as well as a wide range of applications, it may be useful to recognize that some involve imaging, some involve analysis, and some involve a combination of analysis and imaging.

Other embodiments are within the scope of the following claims and other claims.

Claims (19)

1. An apparatus for sample processing for a microscope, comprising:

a light sensitive imaging sensor having a sensor surface for receiving a fluid sample such that the fluid sample is in contact with the sensor surface (103);

a body that moves relative to the light sensitive imaging sensor, the body comprising an extension member having a body surface that contacts a portion of the fluid sample; and

a carrier having an extension (1124), wherein the body is configured to be initially placed on the extension (1124) to enable the body to rest on the extension (1124) of the carrier, the extension member protruding towards the fluid sample, wherein the carrier is configured to be moved towards the fluid sample and towards a sensor surface of the light sensitive imaging sensor to enable a body surface of the extension member to contact a portion of the fluid sample, such that when the body surface of the extension member contacts the portion of the fluid sample, the body is able to rest on the fluid sample independently of movement of the carrier rather than remaining resting on the carrier (1120).

2. The apparatus of claim 1, wherein the body allows light to pass onto the light sensitive imaging sensor.

3. The apparatus of claim 1, wherein a sensor surface of the light sensitive imaging sensor for receiving a fluid sample comprises a hydrophilic coating.

4. The apparatus of claim 1, wherein a surface of the body that contacts a portion of the fluid sample comprises a hydrophilic coating.

5. The apparatus of claim 1, further comprising a sample delivery component for preparing and delivering a fluid sample to a sensor surface of the light sensitive imaging sensor.

6. The apparatus of claim 5, wherein the sample transport component comprises at least two volume capillaries, a nozzle for mixing fluids within the at least two volume capillaries, and an output tip through which a fluid sample is transported to a sensor surface of the photosensitive imaging sensor.

7. The apparatus of claim 1, wherein the body is configured to be initially loosely attached to the carrier.

8. The device of claim 1, wherein the body comprises one or more elements configured to guide motion of the body as the carrier moves toward a sensor surface of the light sensitive imaging sensor.

9. The apparatus of claim 1, further comprising means for adjusting a vertical distance between a surface of the body contacting a portion of the fluid sample and a sensor surface of the light sensitive imaging sensor receiving the fluid sample.

10. The apparatus of claim 1, wherein the body comprises a chamber top.

11. The apparatus of claim 1, wherein the body comprises a lid.

12. The apparatus of claim 1, wherein the carrier is driven by one or more springs.

13. The apparatus of claim 1, wherein the carrier is magnetically actuated.

14. The apparatus of claim 1, wherein the carrier comprises a slot configured to hold the body.

15. The apparatus of claim 1, wherein the extension member comprises a truncated pyramid.

16. A method for sample processing for a microscope, comprising:

supporting a body on a carrier (1120), the body comprising an extension member having a body surface; and

moving the carrier towards a fluid sample on a sensor surface of a light sensitive imaging sensor, the extension member protruding towards the fluid sample to move the body from a position in which the body does not contact the fluid sample to a position in which a body surface of the extension member contacts the fluid sample, such that when the body surface contacts the fluid sample, the body surface is parallel to the sensor surface of the light sensitive imaging sensor, the body rests on the fluid sample, and the body becomes supported by the fluid sample independent of further movement of the carrier.

17. The method of claim 16, wherein supporting a body on a carrier comprises resting the body on a carrier such that a center of the body is vertically aligned with a center of the light sensitive imaging sensor.

18. The method of claim 16, wherein moving the carrier toward the fluid sample comprises displacing a portion of the fluid sample from a sensor surface based on a force applied to the fluid sample by the body.

19. The method of claim 16, wherein the extension member comprises a truncated pyramid.