Method of phase contrasting
11619581 · 2023-04-04
Assignee
Inventors
- Garth Jason Simpson (Lafayette, IN, US)
- Fengyuan Deng (Watertown, MA, US)
- Changqin Ding (West Lafayette, IN, US)
- Chen Li (West Lafayette, IN, US)
Cpc classification
International classification
Abstract
A system including a first micro-retarder array, wherein the first micro-retarder array is configured to convert a purely polarized light of an incident light into two components. The system additionally includes an optical device, wherein the optical device is configured to collimate the two components to two foci planes. Moreover, the system includes a second micro-retarder array, wherein the second micro-retarder array is configured to combine a set of two components of the incident light, thereby producing a second purely polarized light. Further the system includes a detector, wherein the detector is configured to receive the second purely polarized light.
Claims
1. A method for phase contrasting, wherein the method comprises: converting a purely polarized light of an incident light into two orthogonal components, wherein the two orthogonal components comprises: a first purely polarized divergent component and a second purely polarized convergent component, wherein the first purely polarized divergent component is orthogonal to the second purely polarized convergent component; focusing each of the first purely polarized divergent component and the second purely polarized convergent component into a focal plane, thereby producing axially offset two foci planes, placing a first portion of a sample at one focus plane of the two foci planes; combining a phase altered purely polarized light component with a phase unaltered purely polarized light component to form a second purely polarized light; detecting the second purely polarized light by a channeled detector; and calculating a first phase change of the first portion of the sample from the second purely polarized light.
2. The method of claim 1, further comprising: placing a second portion of the sample at the one focus plane of the two foci planes; combining a second phase altered purely polarized light component with a second phase unaltered purely polarized light component to form a third purely polarized light, wherein the second phase altered purely polarized light component comprises a third phase shifted divergent purely polarized light or a fourth phase shifted convergent purely polarized light, wherein the third phase shifted divergent purely polarized light is orthogonal to the fourth phase shifted convergent purely polarized light; detecting the third purely polarized light by the channeled detector; and calculating a second phase change of the second portion of the sample from the third purely polarized light.
3. The method of claim 1, wherein the incident light is emitted from a laser device, an LED, or any light source.
4. The method of claim 1, wherein the converting, and the combining is performed by a micro-retarder array.
5. The method of claim 1, wherein the focusing is performed by an optical device.
6. The method of claim 1, wherein the channeled detector comprises a single channel photo-diode, a single channel photodiode connected to a lock-in amplifier, a lock-in camera, or a camera.
7. The method of claim 4, wherein the combining is performed by a micro-retarder array.
8. The method of claim 2, wherein the channeled detector comprises a single channel photo-diode, a lock-in amplifier, a lock-in camera, or a camera.
9. The method of claim 7, wherein the micro-retarder array comprises: a patterned substrate, wherein the patterned substrate comprises a liquid crystal polymer.
10. The method of claim 9, wherein the liquid crystal polymer comprises a half-wave retardance material.
11. The method of claim 9, wherein the patterned substrate comprises a plurality of concentric circles.
12. The method of claim 11, wherein each concentric circle of the plurality of concentric circles is a different orientation of a fast axis of a half-wave retardance material.
13. The method of claim 1, wherein the purely polarized light comprises a linearly polarized light.
14. The method of claim 13, wherein the two orthogonal components comprises a divergent right circular polarized light component and a converging left circular polarized light component.
15. A system comprising: a first micro-retarder array configured to convert a purely polarized light of an incident light into two components; an optical device configured to collimate the two components to two foci planes; a second micro-retarder array configured to combine a set of two components of the incident light, thereby producing a second purely polarized light; a detector; a light source; a half-wave plate, wherein the light source is configured to transmit the incident light to the half-wave plate; a photoelastic modulator; and a quarter-wave plate, wherein the photoelastic modulator, the half-wave plate, and the quarter-wave plate are configured to produce the purely polarized light from the incident light.
16. The system of claim 15, further comprising: a second optical device, wherein the second optical device is configured to collimate the set of two components of the incident light to the second micro-retarder array.
17. The system of claim 13, further comprising a sample, wherein the sample is placed at a foci plane of the two foci planes.
18. The system of claim 15, wherein a component of the set of two components of the incident light is altered.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
(1) One or more embodiments are illustrated by way of example, and not by limitation, in the figures of the accompanying drawings, wherein elements having the same reference numeral designations represent like elements throughout. It is emphasized that, in accordance with standard practice in the industry, various features may not be drawn to scale and are used for illustration purposes only. In fact, the dimensions of the various features in the drawings may be arbitrarily increased or reduced for clarity of discussion.
(2)
(3)
(4)
(5)
(6)
(7)
(8)
(9)
DETAILED DESCRIPTION
(10) The following disclosure provides many different embodiments, or examples, for implementing different features of the present application. Specific examples of components and arrangements are described below to simplify the present disclosure. These are examples and are not intended to be limiting. The making and using of illustrative embodiments are discussed in detail below. It should be appreciated, however, that the disclosure provides many applicable concepts that can be embodied in a wide variety of specific contexts. In at least some embodiments, one or more embodiment(s) detailed herein and/or variations thereof are combinable with one or more embodiment(s) herein and/or variations thereof.
(11) Axially-offset Differential Interference Contrast Microscopy (ADIC) microscope was constructed based on a bright field microscope frame with the addition of several polarizing optics, as illustrated in
(12) A series of masks with varied thickness were made through photolithography resources within the Birck Nanotechnology Center at Purdue University. Silica beads sized in diameter of 8 μm were used. For quantitative phase imaging, the silica beads were dispersed in nail polish sonically before sealed between cover slip and a glass slide. Mouse tail sections were used as well. Mouse tails were first decalcified in the solution of 23% formic acid, 4% formalin, and 1% methanol for 2 hours, followed by sectioning longitudinally to ensure that sections were retrieved from the central region of the tail. The mouse tail section was then fixed in 10% formalin and embedded in paraffin prior to microtoming into 4 μm thick slides. After sectioning, the mouse tail was stained by hematoxylin and eosin.
(13) The Jones vector describing the detected signal after the sample can be expressed through the Eq (2) as a sum of changes from the two foci. In Eq. (2), t.sub.1 and t.sub.2 are sample transmittances from the two foci separately. δ is the phase change induced by the sample, with opposite signs for RCP and LCP focus planes (sample and reference planes).
(14)
When a half wave plate is inserted to change the incident polarization states, the intensity detected through a polarizer at horizontal polarization state is given by the Eq. (3), in which γ is the rotated angle of half wave plate. The transmittance from the sample is recovered as (t.sub.1+t.sub.2).sup.2.
I(γ)∝t.sub.1.sup.2+t.sub.2.sup.2+2t.sub.1t.sub.2.Math.cos(δ+4γ) (3)
(15) For the LIA detection, PEM and QWP were placed between HWP and beam expansion, the past axis of HWP and QWP were rotated to 22.5° and 45° separately. The detected signal intensity after a polarizer at horizontal polarization state can be written as Eq. (4). As shown in Eq. (5), Δ is the retardance modulation introduced by the PEM as a function of time (τ) with modulation amplitude of 2A. The retardance modulation frequency for PEM was f=50 kHz.
I(τ)∝2(t.sub.1.sup.2+t.sub.2.sup.2)+2t.sub.1t.sub.2.Math.sin(Δ(τ)−δ) (4)
Δ(τ)=2A.Math.sin(2πfτ) (5)
When A is relatively small, the Taylor expansion of Eq. (4) is shown as Eq. (6) with the first seven power series, corresponding to the first four harmonics of the LIA detection. The approximation in Eq. (6) will result in negligible errors for the PEM modulation as long as A<π/2. In our experiments, the modulation amplitude of the PEM was set as A=0.3π.
(16)
(17) According to Eq. (6), quadrature components (sin) only exist for odd harmonic elements while in-phase components (cos) only exist for even harmonic elements of the Taylor series. The quadrature components (sin) of the first harmonic (1f) and the in-phase components (cos) of the second harmonic (2f) LIA detection are written as Eq. (7) and Eq. (8), respectively.
(18)
Combining Eq. (7) and Eq. (8) will result in the quantitative phase shift retrieval function shown below.
(19)
Unlike the HWP rotation strategy, the transmittance image recovered from the LIA detection is defined as t.sub.1t.sub.2 instead of (t.sub.1+t.sub.2).sup.2 since no DC components were collected from the LIA detection. It is noteworthy that the HWP rotation strategy can recover phase value in the range of [−π,π) while the LIA detection can only retrieve phase shift in the range of (−π/2,π/2) due to the limitation of inverse tangent.
(20) We first performed 3D raster scans of a 1951 USAF resolution test chart to demonstrate the ability of the μRA to generate two axially-offset foci once installed in an optical microscope. Comparing to the result obtained from the bright field microscope, the 3D image stack acquired with the μRA shows two crisp images corresponding to the axially-offset dual-foci among slightly blurred images at the rest planes, as illustrated in
(21) To retrieve the quantitative phase contrast images from ADIC imaging using the HWP rotation strategy, we measured a whole set of images with the HWP rotating through a 90-degree range while PD detecting at the horizontal orientation. The rotation interval was set as 3 degree such that 30 γ-x-y images were obtained prior to the phase retrieval process. Then the pixel-by-pixel nonlinear fit was applied to the intensity trace as a function of γ on each pixel referring to Eq. (3). The quantitative phase images were retrieved as the δ map while the transmittance images were recovered as (t.sub.1+t.sub.2).sup.2.
(22) To demonstrate our method with rapid LIA detection, we fed both the 1f (50 kHz) and 2f (100 kHz) reference signal generated by the PEM to the LIA and acquired the ADIC images of the same FoV at two frequencies separately. The raw images (cos components and sin components) collected from 1f and 2f LIA measurements were shown in
(23) The agreement between the two QPI strategies applied to the ADIC microscopy, HWP rotation and LIA detection were tested by imaging the same FoV of a mouse tail section and dispersed 8 μm silica beads shown as
(24) Due to the different equations used for quantitative phase calculation, the recovered phase ranged in [−,π) for HWP rotation strategy and (−π/2, π/2) for LIA detection strategy separately, as discussed in the above paragraphs. The limits of detection (LoD) for the two strategies of ADIC microscopy were provided by retrieving quantitative phase contrast images of background without samples as shown in [
(25) Due to the different recovered phase ranged for HWP rotation and LIA detection strategies, QPI images recovered from the two strategies showed different phase wrapping influences.
(26) Example 1: A method for phase contrasting, wherein the method includes converting a purely polarized light of an incident light into two orthogonal components, wherein the two orthogonal components includes a first purely polarized divergent component and a second purely polarized convergent component. The first purely polarized divergent component is orthogonal to the second purely polarized convergent component. The method additionally includes focusing each of the first purely polarized divergent component and the second purely polarized convergent component into a focal plane, thereby producing axially offset two foci planes. Furthermore, the method includes placing a first portion of a sample at one focus plane of the two foci planes. Additionally, the method includes combining a phase altered purely polarized light component with a phase unaltered purely polarized light component to form a second purely polarized light, wherein the phase altered purely polarized light component includes a first phase shifted divergent purely polarized light component or a second phase shifted convergent purely polarized light component. The first phase shifted divergent purely polarized light component is orthogonal to the second phase shifted convergent purely polarized light component. Moreover, the method includes detecting the second purely polarized light by a channeled detector. Furthermore, the method includes calculating a first phase change of the first portion of the sample from the second purely polarized light.
(27) Next, the method includes placing a second portion of the sample at the one focus plane of the two foci planes. Additionally, the method includes combining a second phase altered purely polarized light component with a second phase unaltered purely polarized light component to form a third purely polarized light, wherein the second phase altered purely polarized light component includes a third phase shifted divergent purely polarized light or a fourth phase shifted convergent purely polarized light. The third phase shifted divergent purely polarized light is orthogonal to the fourth phase shifted convergent purely polarized light. Moreover, the method includes detecting the third purely polarized light by the channeled detector. Further, the method includes calculating a second phase change of the second portion of the sample from the third purely polarized light.
(28) In accordance with the above example, the incident light is emitted from a laser device, an LED, or any light source. The converting, and the combining is performed by a micro-retarder array. The focusing is performed by an optical device. The channeled detector comprises a single channel photo-diode, a single channel photodiode connected to a lock-in amplifier, a lock-in camera, or a camera. The micro-retarder array includes a patterned substrate, wherein the substrate includes a liquid crystal polymer. The liquid crystal polymer comprises a half-wave retardance material. The patterned substrate comprises a plurality of concentric circles. Each concentric circle of the plurality of concentric circles is a different orientation of a fast axis of a half-wave retardance material.
(29) Example 2: A system including a first micro-retarder array, wherein the first micro-retarder array is configured to convert a purely polarized light of an incident light into two components. The system additionally includes an optical device, wherein the optical device is configured to collimate the two components to two foci planes. Moreover, the system includes a second micro-retarder array, wherein the second micro-retarder array is configured to combine a set of two components of the incident light, thereby producing a second purely polarized light. Further the system includes a detector, wherein the detector is configured to receive the second purely polarized light. The system includes a light source and a half-wave plate. The light source is configured to transmit the incident light to the half-wave plate. The half-wave plate is configured to produce the purely polarized light from the incident light. Additionally, the system includes a photoelastic modulator and a quarter-wave plate, wherein the photoelastic modulator, the half-wave plate, and the quarter-wave plate are configured to produce the purely polarized light from the incident light. The system also includes a second optical device, wherein the second optical device is configured to collimate the set of two components of the incident light to the second micro-retarder array.
(30) Upon operation, a sample is placed at a foci plane of the two foci planes. A component of the set of two components of the incident light is altered. The purely polarized light includes a linearly polarized light. And, the two orthogonal components comprises a divergent right circular polarized light component and a converging left circular polarized light component.
(31) Although the present disclosure and its advantages have been described in detail, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the disclosure as defined by the appended claims. Moreover, the scope of the present application is not intended to be limited to the particular embodiments of the process, design, machine, manufacture, and composition of matter, means, methods and steps described in the specification. As one of ordinary skill in the art will readily appreciate from the disclosure, processes, machines, manufacture, compositions of matter, means, methods, or steps, presently existing or later to be developed, that perform substantially the same function or achieve substantially the same result as the corresponding embodiments described herein may be utilized according to the present disclosure. Accordingly, the appended claims are intended to include within their scope such processes, machines, manufacture, compositions of matter, means, methods, or steps.
(32) While several embodiments have been provided in the present disclosure, it should be understood that the disclosed systems and methods might be embodied in many other specific forms without departing from the spirit or scope of the present disclosure. The present examples are to be considered as illustrative and not restrictive, and the intention is not to be limited to the details given herein. For example, the various elements or components may be combined or integrated in another system or certain features may be omitted, or not implemented.