TW201702688A - Optical coherence tomography apparatus and its application - Google Patents

Abstract

Provided herein are devices and systems that apply full-field optical coherence tomography (OCT) technology to three-dimensional skin tissue imaging. A special designed Mirau type objective and an optical microscope module allowing both OCT mode and orthogonal polarization spectral imaging (OPSI) mode are disclosed.

Description

Optical coherence tomography equipment and its application

The present invention relates to an optical coherence tomography apparatus and its use for imaging tissue samples.

Optical coherence tomography (OCT) is a technique for performing high-resolution cross-sectional imaging that provides images of tissue structure (eg, skin tissue) at the micrometer scale. The OCT method measures the interior of the light scattering sample along the OCT beam.

Mohs microscopy surgery is performed under microscopic control from the patient's body for complete excision of basal cell carcinoma (BCC), squamous cell carcinoma (SCC), and less common types of skin cancer. The excised tissue sample (ie, the biopsy) is cut horizontally to provide a tissue section, which is then histologically prepared on a slide. The slides were examined under a microscope to determine if the tumor was completely contained in the resected tissue - this was determined by detecting the absence of a tumor on the edge or edge of the excised tissue. If the tumor is not completely contained in the excised tissue, additional tissue must be resected from the patient's body until all tissue sections taken show that the tumor has been removed from the patient's body. Biological specimen Inspection and histology are the gold standard for tissue diagnosis, so it is generally time consuming for Mohs surgery to require multiple biopsies, so OCT is used to create Mohs microsurgery in an efficient manner. The image of the sample will be very helpful.

The device or system provided by the present invention comprises: a light source module configured to provide light source light to an optical microscope module, the light microscope module uses the light source light and processes the light signal; the Mirau type objective lens module, It utilizes light from the optical microscope module and processes an optical signal generated from a tissue translation module that houses the tissue sample; and a data processing unit including a first detector and a second detector for analyzing the source An optical signal of the tissue sample, wherein the Mirau type objective lens module includes an interference objective lens immersed in the medium, and wherein the optical microscope module includes a first polarization beam splitter and a second polarization beam splitter, the first A polarizing beam splitter is adapted to cause only a portion of the signal light to be orthogonally polarized and projected to the first detector via the second polarizing beam splitter, the second polarizing beam splitter being adapted to cause only the A portion of the signal light is orthogonally polarized and projected to the second detector.

In another aspect, a method for imaging a tissue sample is provided, the method comprising: imaging a test light emerging from a depth direction of a sample through a device or system as described herein, and a second time from the sample A contrast image of the absorption, dispersion, and/or scattering of the substructure is imaged to provide a dynamic state of the sample.

Incorporation

If all publications, patents and specialties mentioned in this manual are mentioned The application is hereby incorporated by reference in its entirety to the extent of the extent of the disclosure of the disclosure of the disclosure of each of each of each of

110‧‧‧Light source module

120‧‧‧Optical microscope module

130‧‧‧Mirau type objective lens module

140‧‧‧Organization translation module

150‧‧‧Data Processing Unit

209‧‧‧Multimode fiber

210‧‧‧Light source module

211‧‧‧Single-clad crystal fiber

212‧‧‧Laser diode

213‧‧‧First Collimation and Focusing Module

214‧‧‧Second collimation and focusing module

220‧‧‧Optical microscope module

221‧‧‧ objective lens

222‧‧‧Optical long wave pass filter

223‧‧‧First beam splitter

225‧‧‧Mirror

226‧‧‧Projection lens

227‧‧‧Second beam splitter

230‧‧‧Mirau type objective lens module

231‧‧‧z-axis piezoelectric transducer (PZT)

233‧‧‧Interference objective

234‧‧‧ objective lens

235‧‧‧Ring container

236‧‧‧First glass plate

237‧‧‧second glass plate

240‧‧‧Organization translation module

241‧‧‧ coverslips

250‧‧‧Data Processing Unit

251‧‧‧First detector

252‧‧‧Second detector

308‧‧‧ imaging fiber bundle

310‧‧‧Light source module

320‧‧‧Optical microscopy module

327‧‧‧Optical focusing mirror

330‧‧‧Mirau type objective lens module

340‧‧‧sample translation module

The features and advantages of the present invention will become more apparent from the detailed description of the embodiments of the invention.

1 is a block diagram representing a device/system of the present invention including a light source module, an optical microscope module, a Mirau type objective lens module, a tissue translation module, and a data processing unit.

2 is another aspect of the apparatus/system of the present invention that provides both an OCT mode and an OPSI mode.

3 is a schematic diagram of a variation of the exemplary inventive device/system of FIG.

4 is a schematic illustration of an exemplary Mirau type objective lens module.

Figure 5 is a radiation spectrum of an exemplary source Ce ³⁺ :YAG single-clad crystal fiber, with an inset showing an end view of the crystal fiber.

Figure 6 shows the optical path difference between water and glass plates measured with one pixel of the CCD.

Figure 7 shows a lateral scan in water showing a lateral resolution of 0.56 μm.

In recent years, optical coherence tomography (OCT) has been widely applied to three-dimensional (3-D) image reconstruction of skin tissue. It is well known that the layer parameters (LP) of the stratum corneum (SC), such as the mean total thickness (a-TT), the average number of layers (a-NOL), and the average cell layer thickness (a-CLT), are non-invasively explored in the epidermis. Etc. It is important to evaluate the skin moisture retention of the epidermis. However, in order to apply OCT technology to skin tissue imaging, an axial resolution better than 1.2 μm in tissue is the threshold for measuring the LP of SC. In addition, the morphology of individual 3-D epidermal cells is also important for early detection of normal and abnormal cells in precancerous diagnosis. These all require sub-micron spatial resolution in the tissue. The full field OCT (FF-OCT) using a two-dimensional CCD/CMOS camera has the opportunity to observe the layer structure of the SC, especially for front side monitoring. Generally, the detection sensitivity of the FF-OCT using a CCD/CMOS camera is about 80 dB, which is related to the camera area size and the en face frame rate.

Keratinocytes and melanocytes are the two major cell types in the epidermis, with a normal size of 10 to 50 μm. Through the keratinization process in about one month, the skin can be divided into several layers, namely the base layer of the bottom, the spinous layer, the granular layer, the transparent layer and the SC at the top. In the epidermis, melanocytes are interspersed in the basal layer and have elongated dendrites. In skin care, proliferation and differentiation of keratinocytes affect epidermal water retention and dry skin diseases.

Apparatus and systems for applying OCT techniques (eg, FF-OCT) to skin tissue imaging are provided herein. In particular, the present invention provides in vitro and in vivo 3-D imaging of skin tissue.

In some embodiments, a device is provided, the device comprising: a light source module configured to provide light source light to an optical microscope module, the light microscope module applying the light source light and processing the light signal; An objective lens module that uses light from the optical microscope module, And processing an optical signal generated from a tissue translation module that houses the tissue sample; and a data processing unit including a first detector and a second detector for analyzing an optical signal from the tissue sample, wherein the Mirau type objective mode The set includes an interference objective immersed in the medium, and wherein the optical microscope module includes: a first polarizing beam splitter adapted to cause only a portion of the signal light to be orthogonally polarized and projected to the first via the second polarizing beam splitter A detector, and a second polarizing beam splitter adapted to only polarize a portion of the signal light and project it to the second detector.

In some embodiments, the light source module includes a spontaneous emission source, an amplified spontaneous emission source, a super luminescent diode, a light emitting diode (LED), a broadband supercontinuum source, a mode-locked laser, and a tunable mine. Emitter, Fourier domain mode-locked source, optical parametric oscillator (OPO), halogen lamp, or doped crystal fiber, such as Ce ³⁺ :YAG crystal fiber, Ti ³⁺ :Al ₂ O ₃ crystal fiber, Cr ⁴⁺ : YAG crystal fiber, etc. In some embodiments, the light source module comprises a Ce ³⁺ :YAG crystal fiber, a Ti ³ +:Al ₂ O ₃ crystal fiber, or a Cr ⁴⁺ :YAG crystal fiber. In some embodiments, the light source module comprises a Ce ³⁺ :YAG crystal fiber.

In some embodiments, a Mirau type objective lens module includes an interference objective lens immersed in a medium that is located in a first glass sheet, a second glass sheet in a sealed container filled with one or more media. In certain embodiments, the interference objective lens is immersed in a medium having optical properties similar to the tissue sample to be analyzed. In certain embodiments, the optical property is a refractive index. In certain embodiments, the medium has a refractive index in the range of from about 1.2 to about 1.8. In certain embodiments, the medium has A refractive index in the range of from about 1.3 to about 1.5. In some embodiments, the medium comprises water, eucalyptus oil, ethanol, glycerin, pyrex, clear glass, or plastic having a refractive index ranging from about 1.3 to about 1.5, or a combination thereof. In certain embodiments, the medium comprises water, eucalyptus oil or glycerin. In certain embodiments, the medium comprises water. In certain embodiments, the medium comprises eucalyptus oil. In some embodiments, the one or more media comprises a first medium and a second medium. In certain embodiments, the first medium comprises water and the second medium comprises eucalyptus oil.

In some embodiments, the tissue translation module includes a cover slip and a lateral motorized linear stage located on the tissue container device. In some embodiments, the tissue container device is a slide or cassette. In certain embodiments, the coverslip acts as a tissue holder.

In some embodiments, the data processing unit includes one or more one-dimensional detectors, one or more two-dimensional detectors, optionally coupled computers, or a combination thereof. In certain embodiments, the data processing unit includes two or more two-dimensional detectors. In certain embodiments, the two or more two-dimensional detectors are charge coupled device (CCD), multi-pixel camera, or complementary metal oxide semiconductor (CMOS) camera, or a combination thereof.

In some embodiments, a system or apparatus is provided, comprising: a Ce ³⁺ :YAG crystal fiber/LED light source module configured to provide source light to an optical microscope module, the optical microscope module utilizing the Light source and process the light signal; the Mirau type objective lens module uses light from the optical microscope module and processes the light signal generated from the tissue translation module; and a data processing unit for analyzing the light signal from the tissue sample, Wherein the Mirau type objective lens module comprises an emu oil, and wherein the optical microscope module comprises a quarter-wave plate as an optical switch, the quarter-wave plate being configured for use in an optical coherence tomography (OCT) mode and Switching between orthogonal polarization spectral imaging (OPSI) modes.

In some embodiments, a system or apparatus is provided, comprising: a Ce ³⁺ :YAG crystal fiber/LED light source module configured to provide source light to an optical microscope module, the optical microscope module application Light source light and process light signals; a Mirau type objective lens module that utilizes light from the optical microscope module and processes light signals generated from the tissue translation module; and includes a first detector and a second detector a data processing unit for analyzing an optical signal from a tissue sample, wherein the Mirau type objective lens module includes an interference objective lens immersed in the medium, and wherein the optical microscope module includes a first polarization beam splitter and a second polarization a beam splitter adapted to cause only a portion of the signal light to be orthogonally polarized and projected to the first detector via a second polarization beam splitter, the second polarization beam splitter being adapted to only A portion of the signal light is orthogonally polarized and projected to a second detector.

Referring to FIG. 1 , an exemplary inventive system/device 100 includes a light source module 110 , an optical microscope module 120 , a Mirau type objective lens module 130 , a tissue translation module 140 , and a data processing unit 150 . The light module 120 is configured to provide appropriate light to the optical microscope module 120, which utilizes source light and processes the light signals. The optical microscope module 120 is associated with the Mirau type objective lens module 130, and the Mirau type objective lens module 130 Light is further processed and injected into tissue samples located in tissue translation module 140. Light returned from the tissue translation module is directed to the data processing unit 150.

In some embodiments, the light source module includes a spontaneous emission source, an amplified spontaneous emission source, a super luminescent diode, a light emitting diode (LED), a broadband supercontinuum source, a mode-locked laser, and a tunable mine. Emitter, Fourier domain mode-locked source, optical parametric oscillator (OPO), halogen lamp, doped crystal fiber (such as Ce ³⁺ :YAG crystal fiber, Ti ³⁺ :Al ₂ O ₃ crystal fiber, Cr ⁴⁺ :YAG Crystal optical fibers, etc., or any other suitable source of light that would be readily recognized by those of ordinary skill in the art to provide suitable light in accordance with the practice of the present invention. In some embodiments, the light source module includes a Ce ³⁺ :YAG crystal fiber, a Ti ³⁺ :Al ₂ O ₃ crystal fiber, or a Cr ⁴⁺ :YAG crystal fiber. For example, the light source module can be a US patent number. One of the light source modules disclosed in U.S. Patent No. 8,416,48, and U.S. Pat.

In some embodiments, the data processing unit includes one or more one-dimensional detectors, one or more two-dimensional detectors, a computer coupled thereto, or a combination thereof. In some embodiments, the data processing unit includes two two-dimensional detectors. The two-dimensional detector may be, for example, a charge coupled device (CCD) or a complementary metal oxide semiconductor (CMOS) camera or the like.

In some embodiments, data processing unit 150 includes two multi-element (ie, multi-pixel) cameras.

2 illustrates an exemplary inventive system/device 200 including a crystal fiber/LED broadband source 210, an optical microscope module 220, and a Mirau type objective module 230 that provide illumination light to an optical microscope module 220 via a multimode fiber 209. The tissue translation module 240 and the data processing unit 250 including the first detector 251 and the second detector 252 are organized. The exemplary light source module 210 includes a Ce ³⁺ :YAG single-clad crystal fiber 211, and the Ce ³⁺ :YAG single-clad crystal fiber 211 is a 1-W, 445-nm laser diode 212 (Nichia), #NDB7875 , Japan) through the first collimation and focusing module 213 (for example, including a 60× aspheric lens, a bandpass filter (Semrock, #FF01-445/45, USA), and a 40× achromatic lens) and a second standard The direct and focus module 214 (eg, including a 40X achromatic objective lens and a 20X achromatic objective lens) is pumped, wherein the function of the band pass filter is to reflect the backward broadband light back to the single-clad crystal fiber 211. The phosphor output from the single-clad crystal fiber 211 is collimated and focused into the multimode fiber 209. The broadband light emerging from the output of the single-clad crystal fiber is coupled into the multimode fiber 209 and then collimated by the objective lens 221 in the optical microscope module 220, wherein the center wavelength and frequency of the light after the single-clad crystal fiber The widths are 560 nm and 95 nm, respectively.

The exemplary optical microscope module 220 includes an objective lens 221, an optical long pass filter 222, and a beam splitter 223. The beam splitter 223 is disposed between the optical long pass filter 222 and the Mirau type objective lens module 230 and guides the light guide. The model is led to the Mirau type objective lens module 230, the mirror 225, and the projection lens 226. In some embodiments, the first beam splitter is a polarizing beam splitter. In some embodiments, an optional use of a polarizing lens is provided between the optical long pass filter and the first beam splitter. Light output from the multimode fiber 209 and reflected by the beam splitter 223 becomes polarized light. The design of the second beam splitter 227 allows this The inventive device/system provides both OCT mode and orthogonal polarization spectral imaging (OPSI) mode. In some embodiments, the second beam splitter is a polarizing beam splitter. In some embodiments, a polarizing lens can be optionally disposed between the first beam splitter and the second beam splitter. The second polarization beam splitter 227 projects the polarized light to the second detector 252. The sample in the tissue translation module 240 and the light beam reflected from the reference arm are directly combined by the first beam splitter 223, and the second polarization beam splitter 227 processes only part of the signal light to be orthogonal. The polarization is projected to the second detector 252 (providing the OPSI mode), and part of the signal light having the same polarization state is reflected by the mirror 225 to the first detector 251 (providing the OCT mode). The system in OPSI mode is capable of detecting the scattered depolarized light in the sample. The OCT mode is particularly effective for imaging samples with deep structures (eg, skin tissue structures). The OPSI mode allows the system of the present invention to detect the absorption, dispersion, and/or scattering contrast images of any substructure or micro-environment of the sample (eg, red blood cells and microvessels). Dynamic state (eg, red blood cells moving in blood vessels). In some embodiments, the data collected from both the first detector and the second detector is further processed by a computer or the like. This exemplary inventive system/device is capable of detecting the overall structure of a tissue sample (e.g., a deep section of a tissue sample) and sub-structures (e.g., red blood cells within an blood vessel), particularly for imaging skin conditions in the body.

After passing through the first polarizing beam splitter 223, the light becomes circularly polarized. The circularly polarized light will become a reverse circular deviation when reflected back from the reference arm and the sample arm by the Mirau type objective lens module 230. Vibration. The beams reflected from the sample and reference arms in the tissue translation module 240 are combined after passing through the polarization beam splitter 223 and then projected onto the second polarization beam splitter 227.

The tissue translation module 240 includes a cover slip 241 that covers a tissue sample (eg, skin tissue) and a lateral motorized linear stage 242 that is positioned on the tissue container device. The tissue container device can be any container suitable for holding tissue. For example, the tissue container device is a slide for holding a biopsy. In some cases, coverslips are used as slides. In certain embodiments, the tissue container device is a coverslip. In some embodiments, the tissue container device is a file for holding a tissue sample, such as a tissue sample from a patient's surgical exposure.

3 provides a variation of the embodiment of FIG. 2 in which imaging fiber bundle 308 is used to transfer light between a Mirau type objective lens module 330 and an optical microscopy module 320, a Mirau type objective lens module 330 and an optical microscopy module. Group 320 processes the light from light source module 310. This variant embodiment provides a mobile/flexible Mirau type objective module 330 for detecting samples in the sample translation module 340. To accommodate this design, an optional collimating lens can be used in the imaging fiber bundle 308 to further align the light to the Mirau type objective lens module. As shown in FIG. 3, the quality of the image is further enhanced by using an optical focusing mirror 327.

Referring to FIG. 4, there is shown an exemplary Mirau type objective lens module of FIG. 2, the Mirau type objective lens module 230 including a z-axis piezoelectric transducer (PZT) 231, the z-axis piezoelectric transducer (PZT) 231 It is optically coupled to a 2D xy linear stage (not shown) and an interference objective 233. In detail, this specially designed The Mirau type interference objective 233 includes an objective lens 234 (for example, Olympus, LUMPLFLN 20×W, NA: 0.5, field of view: 550 μm, Japan) immersed in a first medium (for example, water), a ring receiver 235, for Two fused silica glass plates (thickness: 150 μm, λ/10 flat, first glass plate 236 and second glass plate 237) containing a second medium (for example, eucalyptus oil). The diameter of the water focal field is about 220 μm (using 1/3 field of view). The interference objective 233 is fixed to the z-axis piezoelectric transducer 231 (PI, #P-720, Germany). In some embodiments, the first medium is the same as the second medium, for example, the first medium and the second medium may both be eucalyptus oil.

A cover glass 241 is laminated under the sample. In some embodiments, the coverslip has the same thickness as the glass sheet. The total optical travel of the PZT with open loop control is 112 μm. A 500 μm diameter black ink absorber (n=1.48) on the same plane as the objective lens 234 is used to match the refractive index of the first glass plate 236 to absorb stray light back to the data processing unit (ie, CCD) and for Eliminate the DC term of the intensity. After coating the staggered layers with TiO ₂ /SiO ₂ (T/R=60/40, T: transmittance; R: reflectance; n _{矽 oil} = 1.406), a wide frequency _band is coated on top of the second glass plate 237. Beam coat coating. The reflective coating of the first glass sheet 236 that contacts the second medium (i.e., eucalyptus oil) is about 4% due to n _{矽 oil} = 1.406.

During operation, the objective lens 234 directs light through the first glass sheet 236 to focus the test sample on the tissue translation module 240. The second plate 237 reflects the first portion of the focused light to the first glass plate reflective coating to define a reference light and transmits the second portion of the focused light to the test sample to define the measurement light (measurement light) ). Then, the first The second plate 237 recombines the measurement light reflected (or scattered) from the test sample with the reference light reflected from the reflective coating on the first glass plate, and images the combined light by the objective lens 234 and the imaging lens to Interference is generated on processing unit 250 (e.g., a multi-pixel camera with or without a computer). The PZT 231 is coupled to a 2D x-y linear platform 232.

In some embodiments, the interference objective 233 includes an objective lens 234 immersed in the medium, a first glass plate 236, and a second glass plate 237 located in a sealed container filled with one or more media. The medium described herein is defined as any medium having characteristics that compensate for dispersion in the optical path introduced by the beam passing through the medium. The one or more media in the Mirau type objective module provide a means of reducing dispersion in the case of tomographic imaging as compared to conventional Mirau objectives filled with air. In some embodiments, the Mirau type objective lens module of the present invention includes two or more media (eg, a first medium, a second medium, etc.), wherein at least one of the media has optical characteristics similar to those of the sample to be analyzed, It is arranged to compensate for the dispersion in the optical path introduced by the beam passing through the Mirau type objective. Among various skin optical properties, refractive index is an important one. On a microscopic scale in the range of 1 to 10 μm, a change in refractive index causes light scattering. The refractive index of human skin tissue has been determined based on known methods (for example, Ding et al., Physics in Medicine and Biology, 2006, 51(6), 1479). The refractive index of human skin tissue is about 1.38 to 1.44 compared to the refractive index of 1.00 of air at standard temperature and pressure (STP). The medium having a refractive index of between 1.3 and 1.5 is not excluded, for example, including water (1.33), eucalyptus oil (1.336-1.582, depending on composition), 20% aqueous glucose solution (1.36), ethanol (1.36), glycerin (1.47), Pyrex glass (1.47). In some embodiments, the medium used in the Mirau type objective lens module has a refractive index ranging from about 1.0 to about 2.0, from about 1.2 to about 1.8, from about 1.3 to about 1.6, or from about 1.3 to about 1.5. In certain embodiments, the medium has a refractive index ranging from about 1.3 to about 1.5.

For example, the objective lens 234 is immersed in water (ie, a first medium) or a liquid having optical characteristics close to the optical characteristics of water. This is because the sample to be imaged (eg, living cells, skin tissue) contains the most water. Thereby, imaging of living cells can be performed in a satisfactory manner. In some embodiments, the one or more media are liquids, gels, special glasses, special plastics, or any other suitable material having optical properties that are close to the optical properties of the test sample. In certain embodiments, the medium is a liquid. In some embodiments, the liquid medium includes water, glycerin, ethanol, eucalyptus oil, and the like. In certain embodiments, the liquid medium comprises water. In certain embodiments, the liquid medium comprises eucalyptus oil. In certain embodiments, the liquid medium comprises glycerin. In some embodiments, the medium is a clear glass or plastic having a refractive index in the range of from about 1.3 to about 1.5.

As shown in Figure 2, PZT 231 is biased by an amplified signal from a DAQ card (NI, #PCI-4461, USA) with open loop mode. The z-axis position and input voltage of the PZT are recorded by the Michelson interferometer through the count wavenumber and phase difference of the He-Ne laser. Therefore, the hysteresis motion of the PZT is experimentally compensated by the recorded position-voltage curve. For example, Figure 5 is the emission spectrum of Ce ³⁺ :YAG SCF (exemplary light source), with the inset showing the end view of the SCF. The interferometric signal intensity of the A-scan reflected from the boundary between the water and the glass plate is measured by one pixel of the CCD (see Figure 6). The carrier envelope strength from the carrier signal in Figure 5 is calculated after the bandpass filter and the Hilbert transform. The detection sensitivity calculated by the known method is about 81 dB. The noise floor of the system of the present invention is substantially suppressed by the stronger confocal gate (e.g., NA: 0.8 versus 0.5) effect, and in turn further reduces the ghost effect. An exemplary interference objective correspondingly provides Ra = 0.91 μm (see Figure 5) and Rt = 0.56 μm (see Figure 7) in the axial and transverse directions on the surface of the aqueous medium (or on the SC surface (after water binding, n = 1.47), experimental resolution of Ra = 0.90 μm and Rt = 0.51 μm; however, according to Equation 1, the theoretical spatial resolution at the water surface that satisfies the diffraction limit is Ra = 0.56 μm and Rt = 0.43 μm (or in SC) Surface, Ra = 0.55 μm and Rt = 0.39 μm).

Wherein △ Z _eff is meant a _confocal △ Z (confocal water gate (confocal gate), _{the water} is equal to λ ₀ n / NA ^2, for a 40 × objective (NA: 0.8) of about 1.16μm) and △ Z _homology ( The coherent gate in water is equal to 0.44 λ ₀ ² /n _water Δλ, and the effective axial resolution contributed to the Ce ³⁺ :YAG source with the same objective lens is about 1.09 μm). The n sample and n water are correspondingly the refractive indices of the sample and water. λ ₀ and Δλ are the center wavelength and bandwidth of the light source. In Fig. 4, a 40x interference objective lens 234 is used for water immersion. Surprisingly, it has been found that when a 20x interference objective lens (where NA is 0.5) is used, sample scanning becomes more efficient but still achieves similar 3-D imaging results (eg, resolution). Thus, in some embodiments, the objective lens used in the device/system of the present invention has an NA of 0.5 or less. Since both water and eucalyptus oil have similar refractive indices similar to those in the sample tissue, those of ordinary skill in the art will readily appreciate mutual substitution, or use only water, or only eucalyptus oil, or in accordance with implementations of the present invention. Use any other suitable media. For example, an interference objective lens immersed in an eucalyptus oil (first medium) and a second medium eucalyptus oil is prepared to overcome the evaporability of the water-based Mirau type objective lens module.

Typically, FF-OCT uses a single-shot CCD at 0°, 90°, 180°, and 270° to obtain a frontal image by calculating stack information via a phase step technique, where the triangle oscillations through PZT Exercise to phase shift it. As the exposure time of a front image increases, the detection sensitivity becomes better. Then, the 3-D image is reconstructed by stacking the frontal image along the z-axis. Unlike the classic FF-OCT, the inventive apparatus/system including the Mirau type objective lens reconstructs a 3-D image stack by sequential interferometric signals. This secondary consideration will result in better depth imaging of the device/system of the present invention.

The device or system of the present invention is capable of imaging tissue samples efficiently and easily, and is particularly useful in assisting skin treatment, for example, the device or system of the present invention can be used to assist in Mohs surgery. During the surgical procedure, after each removal of the tissue, when the patient is waiting, the tissue can be examined for cancer to determine if additional tissue removal is required. Mohs outside Surgery is one of the many ways to achieve complete edge control during skin cancer removal, depending on the complete perimeter edge and deep edge assessment. The device or system of the present invention can image tissue tissue in situ or after removing tissue tissue from the patient's body, thereby providing an effective means of assisting Mohs surgery. In some embodiments, a method for imaging a tissue sample is provided, the method comprising: imaging a test light emerging from a depth direction of a sample through the apparatus or system described herein, and from the sample The contrast image of the absorption, dispersion, and/or scattering of the secondary structure is imaged to obtain the dynamic state of the sample. In some embodiments, the tissue sample is skin tissue. In certain embodiments, the method can be used to image skin tissue conditions. In certain embodiments, skin conditions are determined through a complete peripheral perimeter and deep edge assessment.

Although the preferred embodiment of the present invention has been shown and described, it is to be understood by those of ordinary skill in the art that Various changes, modifications, and substitutions may be made by those skilled in the art, and it is understood that the invention may be practiced with various alternatives to the embodiments of the invention described herein. The scope of the invention is intended to define the scope of the invention, and to cover the methods, structures, and equivalents thereof.

209‧‧‧Multimode fiber

210‧‧‧Light source module

211‧‧‧Single-clad crystal fiber

212‧‧‧Laser diode

213‧‧‧First Collimation and Focusing Module

214‧‧‧Second collimation and focusing module

220‧‧‧Optical microscope module

221‧‧‧ objective lens

222‧‧‧Optical long wave pass filter

223‧‧‧First beam splitter

225‧‧‧Mirror

226‧‧‧Projection lens

227‧‧‧Second beam splitter

230‧‧‧Mirau type objective lens module

231‧‧‧z-axis piezoelectric transducer (PZT)

233‧‧‧Interference objective

240‧‧‧Organization translation module

241‧‧‧ coverslips

250‧‧‧Data Processing Unit

251‧‧‧First detector

252‧‧‧Second detector

Claims (27)

A device comprising: a light source module configured to provide a source of light to an optical microscope module, the optical microscope module utilizing the source light and processing an optical signal; a Mirau type objective lens module Using light from the optical microscope module and processing an optical signal generated from a tissue translation module housing a tissue sample; and a data processing unit including a first detector and a second detector For analyzing an optical signal from the tissue sample, wherein the Mirau type objective lens module includes an interference objective lens, the interference objective lens is immersed in the medium, and wherein the optical microscope module includes a first beam splitter and a a second beam splitter adapted to cause only a portion of the signal light to be orthogonally polarized and projected to the first detector via the second beam splitter, the second beam splitter The device is adapted to only polarize a portion of the signal light and project it to the second detector. The device of claim 1, wherein the light source module comprises a spontaneous emission light source, an amplified spontaneous emission light source, a super-radiation light-emitting diode, a light-emitting diode (LED), a broadband super-continuous spectrum light source, Mode-locked laser, tunable laser, Fourier-domain mode-locked source, optical parametric oscillator (OPO), halogen lamp, Ce ³⁺ :YAG crystal fiber, Ti ³⁺ :Al ₂ O ₃ crystal fiber, Cr ^{4 +} : YAG crystal fiber. The device of claim 3, wherein the light source module comprises a Ce ³⁺ :YAG crystal fiber, a Ti ³⁺ :Al ₂ O ₃ crystal fiber, or a Cr ⁴⁺ :YAG crystal fiber. The device of claim 3, wherein the light source module comprises a Ce ³⁺ :YAG crystal fiber. The apparatus of claim 1, wherein the Mirau type objective lens module comprises an interference objective lens immersed in a medium, a first glass plate in a sealed container filled with one or more mediums, and a Second glass plate. The device of claim 1, wherein the interference objective lens is immersed in a medium having optical properties similar to the tissue sample to be analyzed. The device of claim 6, wherein the optical property is a refractive index. The device of claim 7, wherein the medium has a refractive index in the range of from about 1.2 to about 1.8. The device of claim 8, wherein the medium has a refractive index in the range of from about 1.3 to about 1.5. The device of claim 9, wherein the medium comprises water, eucalyptus oil, ethanol, glycerin, pyrex glass, transparent glass or plastic having a refractive index in the range of from about 1.3 to about 1.5, or combination. The device of claim 10, wherein the medium comprises water, eucalyptus oil or glycerin. The device of claim 11, wherein the medium comprises eucalyptus oil. The device of claim 5, wherein the one or more media comprises a first medium and a second medium. The device of claim 1, wherein the first medium comprises water and the second medium comprises eucalyptus oil. The device of claim 1, wherein the optical microscope module further comprises an objective lens, an optical long pass filter, and a polarization beam splitter. The device of claim 1, wherein the tissue translation module comprises a cover slip and a lateral motorized linear stage located on a tissue container device. The device of claim 16, wherein the tissue container device is a slide or a cartridge. The device of claim 16, wherein the cover slip is used as the tissue container. The device of claim 2, wherein the first detector or the second detector are each independently a one-dimensional detector or a two-dimensional detector, an optionally coupled computer, or a combination thereof . The device of claim 19, wherein the first detector or the second detector is a two-dimensional detector. The device of claim 20, wherein the two-dimensional detector is a charge coupled device (CCD), a multi-pixel camera, or a complementary metal oxide semiconductor (CMOS) camera, or a combination thereof. The device of claim 1, wherein the light source module comprises a Ce ³⁺ :YAG crystal fiber, and the Mirau type objective lens module comprises an eucalyptus oil as one or more media. A system for imaging a tissue sample, comprising: a light source module configured to provide a light source light to an optical microscope module, the optical microscope module applying the light source light and processing the light signal; a Miraau type objective lens module that utilizes light from the optical microscope module and processes an optical signal generated from a tissue translation module that houses a tissue sample; and a data processing unit that includes a first detector And a second detector for analyzing an optical signal from the tissue sample, wherein the Mirau type objective lens module includes an interference objective lens immersed in the medium, and wherein the optical microscope module includes a first polarization a beam splitter and a second polarization beam splitter adapted to cause only a portion of the signal light to be orthogonally polarized and projected to the first detection via the second polarization beam splitter The second polarizing beam splitter is adapted to only polarize a portion of the signal light and project it to the second detector. A method for imaging a tissue sample, comprising: imaging a test light emerging from a depth direction of a sample by the apparatus of claim 1 or the system of claim 23 And imaging the contrast image of the absorption from the substructure of the sample. The method of claim 24, wherein the sample is skin tissue. The method of claim 25, wherein the method is for imaging a skin tissue condition. The method of claim 26, wherein the determining the condition of the skin is assessed by a complete surrounding perimeter and a deep edge.