JP3222366U - Microscopy improvements or microscope-related improvements - Google Patents

Abstract

An object of the present invention is to provide an inverted optical microscope in which an objective lens is firmly installed and has high robustness and stability. An inverted optical microscope includes a housing configured to enclose one or more optical components including an objective lens, and the one or more optical components are firmly installed. And a mounting plate portion 18 of the housing 12 configured to achieve this. The housing is a monolithic housing 12, which is a large rigid block of material such as metal, for example, and has one or more removable panels connected to the mounting plate 18 and / or the sealing plate. [Selection] Figure 1B

Description

  The present invention relates to improvements related to microscopes, and in particular to inverted light microscopes, such as interferometric light scattering microscopes.

  Optical microscopes, such as interference scattering microscopes, have conventionally been used to study the structure and kinetics of biological and / or chemical samples. The technique of Interferometric Scattering Microscopy (iSCAT) relies on collecting the light scattered by the object, with a reference light field provided by reflection at the interface. The total intensity of light returning in the direction of the incident beam in the presence of a scattering object is different from that in the absence of such an object due to the interference of scattered and reflected light. Because of the need to achieve high sensitivity in iSCAT, Applicants are able to use iSCAT to detect, capture and even quantify single molecules such as peptides and DNA molecules. I found that. This detection may be unlabeled.

  Optical microscopes, such as inverted optical microscopes, can be susceptible to external influences such as vibrational disturbances, temperature changes or acoustic coupling, which is the sensitivity required for detection of label-free single molecules. This is particularly problematic when working at levels. Usually, this has been addressed by using expensive and bulky optical tables that use active vibration isolation. However, setting up a bulky optical table can be a time-consuming task, and can lead to very complex solutions where external noise sources can not be completely eliminated. . Furthermore, the extra space required for such instruments is not possible for many laboratories. Therefore, it is desirable to be able to provide an optical microscope as a benchtop device.

  Thus, the art develops a bench top version optical microscope setup with high robustness and stability, with the aim of reducing and / or eliminating vibrations and other disturbances for single molecule measurements. There is a demand for and a need for that.

The present invention is created from such background.
According to the present invention, an inverted light microscope is provided, wherein the inverted light microscope is
A monolithic housing surrounding a plurality of optical components, including an objective lens;
A monolithic housing and an integral mounting plate configured to achieve at least a rigid mounting of the objective lens,
The monolithic housing has a thickness greater than 20 mm.

Alternatively, according to the present invention, an inverted light microscope is provided, wherein the inverted light microscope is
Comprising a monolithic housing surrounding a plurality of optical components, including an objective lens,
The monolithic housing is at least 20 mm thick and comprises at least an installation plate configured to achieve a rigid installation of the objective.

  A housing according to any aspect of the present invention may be provided to partially or totally surround a plurality of optical components, including the objective lens. In some embodiments, the housing may be a 3D housing structure, such as a box, capable of enclosing an optical component, including an objective lens, said objective lens being secured to the mounting plate portion / part of the housing Ru. The housing provided by the present invention can significantly reduce or even eliminate acoustical influences or disturbances, such as vibrations, on optical components located in the housing. This can improve stability and accuracy and allow one to perform single molecule measurements. A monolithic housing can provide a 3D structure with walls surrounding the mounting plate, and at each corner, a radius can be designed to help dampen vibrations of a particular frequency. Applicants contemplate that the 3D structure of the housing will help to shift the natural frequency out of the range of the relevant frequency band.

  The housing is monolithic so that the housing can be formed from one substrate or material. A monolithic housing has or comprises a mounting plate portion. The mounting plate is integral with the housing and is part of the housing. The housing may be described as being made of a single piece of material without seams or joints. In some embodiments, the mounting plate can comprise a plurality of openings.

  Furthermore, the inverted microscope with monolithic housing with installation plate according to the invention enables the light microscope setup to function as a stand-alone bench top unit, which is an environment without sophisticated vibration and temperature control means Can operate at

  In some embodiments, the housing can be configured to eliminate vibration in at least one mode. Furthermore, the housing may be configured to support the mounting plate in a 3D housing structure, which may make it impossible to generate certain vibration modes. In some embodiments, the housing is configured to have a 3D shape that can be used to suppress, reduce or shift the natural frequency.

  In some embodiments, multiple optical components may be attached to the housing via the mounting plate using one or more securing means. The fixing means is advantageous as it provides a means of rigidly fixing the optical component to the mounting plate.

  The fixing means for the optical components, including the objective lens, may be Royal Microscopy Society (RMS) or M27 threads. By using the Royal Microscopy Society (RMS) or M27 threads, the optical components, including the objective lens, can be rigidly fixed on the mounting plate. This is very useful because it prevents relative movement of the objective relative to other optical components placed on the mounting plate. Furthermore, a threaded exact hole for each optical component is fixed in its exact position on the mounting plate.

  The housing can have a thickness of at least 20 mm, or the housing can have a thickness of at least 25 mm, at least 30 mm, at least 35 mm, or at least 40 mm. The mounting plate portions or mounting plate sections of the housing may be configured to have a thickness such that they are staggered relative to the remaining portion of the housing.

  The mounting plate can have a thickness of at least 25 mm, or the mounting plate can have a thickness of at least 30 mm, at least 35 mm, at least 40 mm, at least 45 mm, or at least 50 mm. The thickness of the mounting plate portion of the housing can provide rigidity to the device, which in some embodiments can be supported by the structure of other portions of the housing. In one embodiment, the mounting plate portion forms the top or bottom of the housing and may be surrounded by a wall. These walls can support the mounting plate.

  In some embodiments, the inverted light microscope may further comprise a sample area provided on the mounting plate, and the following: interferometer head, sample section lid, sample holder, sample shutter (sample shutter) And / or an actuator and / or a sensor.

Preferably, the inverted light microscope may further comprise a spatial filter rigidly attached to the mounting plate.
In some embodiments, the inverted light microscope can further comprise a sealing plate. The sealing plate may be particularly useful in sealing the microscope to eliminate air flow and temperature gradients inside the housing. In addition, the sealing plate can further help to reduce the effects of beam perturbations and to slow down the alignment drift.

  An inverted light microscope may comprise one or more removable panels configured to be connected to the mounting plate and / or the sealing plate. A removable panel can be advantageous as it can provide a simple and easy way to access the interior of the housing.

  In some embodiments, the housing may be solid. In some embodiments, the housing is made of metal. In a preferred embodiment, the housing may be aluminum, invar or stainless steel. Preferably, the housing is formed of aluminum. Forming the housing from a single metal block, such as aluminum, can help the aluminum improve stability, thermal performance, and ease of assembly.

In some embodiments, the housing can be configured to provide uniform heat absorption and dissipation.
In some embodiments, the thickness of the mounting plate may exceed 50 mm, 70 mm, 100 mm, 150 mm, 200 mm, 250 mm, or 300 mm.

  In some embodiments, the size of the housing is approximately 300 mm deep by 450 mm wide by 150 mm high. In some embodiments, the housing forms a box around the optical components, wherein the mounting plate portion forms the top or bottom section of the housing.

  Other components necessary for operation of the microscope, such as control means or a power supply, may also be present in the housing or outside the housing. External components can be linked using a central line or umbilical.

  The invention will now be described in more detail, by way of example only, with reference to the accompanying drawings.

FIG. 1A is a CAD diagram showing the housing. FIG. 1B is a CAD diagram showing a mounting plate portion of a housing according to the present invention. FIG. 1D is a CAD drawing showing the housing according to FIGS. 1A and 1B. FIG. 3 is a CAD drawing showing the mounting plate part of the housing according to FIGS. 1A, 1 B and 2; FIG. 4 shows a sample area of the housing according to FIGS. 1A, 1 B, 2 and 3; FIG. 7 is a graph showing FFT analysis of a fast imaged fixed reference marker image sequence when the objective lens is fixed. FIG. 7 is a graph showing an FFT analysis of a series of images of fixed reference markers captured at high speed when the objective is installed in a piezoelectric Z-axis drive.

  With reference to FIGS. 1A and 1B, FIGS. 2 and 3, an inverted light microscope setup 10 is presented. An inverted light microscope comprises a housing 12 configured to surround one or more optical components 14, including an objective lens 16. An inverted light microscope has an installation plate portion 18 of the housing 12 configured to provide a rigid installation of one or more optical components 14.

  The housing is a monolithic housing 12, for example, a large rigid block of material such as metal. As shown in FIGS. 1B, 2 and 3, the housing has one or more removable panels 20 connected to the mounting plate 18 and / or the sealing plate. A sealing plate seals the microscope to eliminate air flow and temperature gradients inside the housing for the purpose of reducing the effects of beam perturbations and reducing the speed of alignment drift. All electronic components can be housed in separate boxes connected via umbilicals including all relevant cables and power supplies. The aim is to keep the inverted microscope instrument according to the invention as compact and portable as possible. In addition, the housing 12 can be provided with one or more holes 21 of any size on the side of the housing, as shown in FIG. 3, the holes 21 being in the housing and / or of the housing It can be used to enable electronic wiring outside. Additionally or alternatively, the holes may be useful for mounting optical components such as cameras.

  Optical components may be enclosed within the housing, and the housing may be made of aluminum, having approximate dimensions of 300 mm deep by 450 mm wide by 150 mm high. The weight of the housing is about 10 kg to 100 kg. In some embodiments, the housing exceeds 10 kg, 20 kg, 30 kg, 40 kg, 50 kg, 60 kg, 70 kg, 80 kg, or 90 kg. In some embodiments, the housing may be less than 100 kg, 90 kg, 80 kg, 70 kg, 60 kg, 50 kg, 40 kg, 30 kg, 20 kg, or 10 kg. Preferably, the housing weighs about 25 kg.

  The thickness of the housing may exceed 20 mm. The thickness of the mounting plate portion may exceed 20 mm, or 50 mm. Thus, the housing can have portions or sections of different thickness.

  The weight and thickness of this housing are inverted as vibration is coupled into the microscope instrument to prevent degradation in data quality during detection and analysis of biological and / or chemical samples such as single molecules. It can be of great importance for the stability and performance of scanning light microscopes. In addition, the housing further reduces the need for expensive and bulky optical tables that use active vibration isolation.

  Referring to FIG. 1B, objective lens 16 and optical components 14 such as a spatial filter are fixed to integral mounting plate 18 by fixing means 22 so that all of these components are mechanically rigidly coupled. Ru. In addition, a mounting pillar 17 for the 45 ° mirror is placed below the objective lens 16. The mounting plate 18 may have a thickness greater than 50 mm. The reason is that this reduces the influence of the vibration of the housing. The fastening means 22 may be the threads of the Royal Microscopy Society screwed into the mounting plate 18 or the threads of the M 27, in which the objective lens is fastened. It is very important to ensure that the objective is maximally stable for the remaining beam path. Optional optical components may be included in the inverted microscope of the present invention. Exemplary components may include illumination sources, detectors, lenses such as condenser lenses and tube lenses, and / or beam splitters. The microscope objective discussed herein may be referred to as an objective lens.

  Referring to FIGS. 1A and 4, a sample area 24 is provided on the mounting plate. The sample area 24 can include one or more of the following: microscope head 26, such as an interferometer head, sample holder 28, sample stage lid 30, actuator, and RFID proximity sensor, for example, for shutter operation etc. One or more sensors 32. The microscope head 26 can have an objective 16. The lid 30 may be a metal such as aluminum. The sample area 24 may be fixed to the mounting plate using fixing means 22.

  An RFID proximity sensor 32 may be connected to a mechanical shutter mounted on the mounting plate. As a possibility, the light source may be a laser. The use of proximity sensors allows the high power laser to be incident on the objective only when the lid completely surrounds the sample area, thereby making the microscope a class 1 laser device.

  Referring to FIGS. 1B and 3, the housing is designed such that each of the optical mounts and optical components will have threaded holes 34 only at the locations required for proper alignment. This improves the ease of manufacture and the alignment of the components. The reason is that the components do not shift, and each optical component has only one place that can be installed on the installation plate.

  The microscope of the present invention may be any inverted light microscope. This may include any super-resolution microscope, optical tweezers setup, or scanning probe microscope such as atomic force microscope. Preferably, the microscope is an interference microscope, such as an interference scattering microscope.

The housing may be referred to as a box or shell that encloses the optical component therein. This box or shell is composed of one substrate. If necessary, additional components such as experimental circuit boards may be included in the housing and additional components may be affixed to the installation plate. The experimental circuit board can be used to secure further additional components.
Example A number 1.5 glass coverslip is washed by rinsing continuously with MilliQ and ethanol and placed on top of a microscope. A fiducial marker (particles of impurities or gold nanoparticles) is moved into the field of view of the microscope and a motion picture is taken at the maximum frame rate allowed by the CMOS imaging camera. The point spread function of the particle is then adapted to a 2D Gaussian function, thereby tracking its position over time. Referring to FIGS. 5 and 6, by performing a Fast Fourier Transform (FFT) of this position trace, any intermittent oscillatory coupling to the setup becomes apparent. This process is then repeated, in which the microscope objective is installed in a piezoelectric Z-axis drive instead of being mechanically coupled to the optics installation plate (see FIG. 5). (See Figure 6). With reference to FIGS. 5 and 6, experiments clearly show the following: When the microscope objective is fixed in the optics installation plate (see FIG. 5), the amplitude decreases and the peak of the FFT The numbers are reduced, which correspond to good attenuation of external vibration and noise sources.

Various other aspects and embodiments of the present invention will be apparent to those skilled in the art from consideration of the present disclosure.
As used herein, “and / or” should be construed as specifically disclosing each of the two specified features or components, whether or not the other is present. For example, “A and / or B”, as if each were individually described herein, may each specifically (i) A, (ii) B, (iii) A and B, Are to be interpreted as being

  Unless stated otherwise by context, the descriptions and definitions of the features described above are not limited to any particular aspect or embodiment of the present invention, but apply equally to all aspects and embodiments described.

  Although the invention has been described with reference to several embodiments by way of example, the invention is not limited to the disclosed embodiments and departs from the scope of the invention as defined in the appended claims. One of ordinary skill in the art will recognize that alternative embodiments can be constructed without.

Claims (15)

An inverted light microscope, wherein the inverted light microscope is
Comprising a monolithic housing surrounding a plurality of optical components, including an objective lens,
Said monolithic housing having a thickness of more than 20 mm, comprising an installation plate configured to realize at least a rigid installation of said objective lens,
Inverted light microscope.   The inverted optical microscope according to claim 1, wherein the plurality of optical components are attached to the mounting plate using one or more fixing means.   The inverted light microscope according to claim 1 or 2, wherein the mounting plate has a thickness of at least 25 mm.   The inverted light microscope according to any one of the preceding claims, wherein the housing is configured to eliminate vibrations in at least one mode.   The apparatus further comprises a sample area provided on the mounting plate, and one or more of the following: an interferometer head, a sample holder, a sample shutter, an actuator, and / or a sensor. The inverted light microscope according to any one of claims 1 to 4.   The inverted optical microscope according to any one of claims 1 to 5, further comprising a spatial filter rigidly attached to the mounting plate.   The inverted light microscope according to any one of the preceding claims, further comprising a sealing plate.   8. An inverted light microscope according to any one of the preceding claims, further comprising one or more removable panels configured to be connected to the housing and / or the sealing plate.   9. An inverted light microscope according to any one of the preceding claims, wherein the housing is rigid.   10. An inverted light microscope according to any one of the preceding claims, wherein the housing is metal.   The inverted light microscope of claim 10, wherein the housing is aluminum, invar, or stainless steel.   An inverted light microscope according to any of the preceding claims, wherein the housing is configured to provide uniform heat absorption and heat dissipation in at least one mode.   13. An inverted light microscope according to any one of the preceding claims, wherein the thickness of the mounting plate is greater than 50 mm.   14. An inverted light microscope according to any one of the preceding claims, wherein the size of the housing is approximately 300 mm deep x 450 mm wide x 150 mm high.   The inverted light microscope according to any one of the preceding claims, wherein the fixing means for the objective lens is a thread of Royal Microscopy Society (RMS) or M27.

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