
- Photonics Research
- Vol. 13, Issue 4, 1049 (2025)
Abstract
1. INTRODUCTION
By detecting the material modulation for the polarization of incident light, the composition and fine structure of the material can be revealed by using specialized optical techniques and computational methods [1,2]. Compared to intensity-based methods, polarization technologies can obtain multi-dimensional characteristics and improve the sensitivity and specificity of material detection. Currently, polarization measurement is attracting increasing attention across various research fields, including material characterization [3], biomedical studies [4], and clinical applications [5].
Interferometric imaging, including optical coherence tomography (OCT) and holographic microscopy, can extract three-dimensional (3D) structures from interference signals. Integrating interferometric imaging with polarization measurement has become an effective approach for obtaining 3D polarization information. Among these methods, polarization-sensitive OCT (PS-OCT) [6–8], as a representative interferometric imaging method with polarization measurement function, can offer high-resolution, non-invasive, and non-destructive imaging [9,10]. By introducing polarization modules, PS-OCT provides a series of polarization parameters related to birefringence and dichroism, demonstrating great potential in ophthalmology [11], cardiology [12], dermatology [13], and more.
With the growing demand for diagnostic applications, polarization measurement methods of interferometric imaging have continually evolved. At present, there are three kinds of methods. (1) Single-input methods [14–17] use the single polarized probe light and can obtain polarization information in a single acquisition. However, this method limits the variety of polarization parameters and may introduce phase jump errors [17]. (2) Multi-input methods [18–20] use multiple polarized probe lights and offer accurate and comprehensive polarization parameters. However, multiple signals are acquired in a time sequence, decreasing the acquisition speed. (3) Single-detector-based methods [21–23] simplify the setup. However, they rely on the time-sequenced or the space-division method, limiting the acquisition speed or the axial imaging range.
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As a representative multi-input method, polarization modulation methods control the polarization state of the probe light by an electro-optic modulator [19,24], obtaining accurate and rich polarization information while reducing the acquisition speed. Modulating polarization in the spectral domain rather than the time domain offers the potential to overcome the limitation of acquisition speed. Currently, this idea is primarily used in polarimetry [25–27] represented by polarization spectral intensity modulation (PSIM). These methods employ wavelength-sensitive birefringent materials and the polarizer to modulate polarization as an intensity function of wavelength, followed by demodulating the spectrum to extract polarization information.
However, integrating PSIM into interferometric imaging has challenges. PSIM causes the polarization to change continuously with the wavelength, which makes it difficult to meet the achromatic polarization state required for current interferometric polarization modulation methods. Besides, PSIM introduces an intensity modulation on the spectrum, which could interfere with depth-resolved imaging. To address these challenges, specific polarization calculation methods and spectral-polarization-modulation (SPM) strategies should be developed to mitigate intensity modulation and ensure compatibility with interferometric imaging.
In this study, we proposed a rapid and simple interferometric polarization measurement method and integrated it into an OCT system to establish an advancing interferometric imaging system called SPM-OCT. This method can extract birefringent and dichroic parameters from the polarization-modulated signal without reducing the acquisition rate or the axial imaging range. The standard polarization elements, a skinned mouse leg, and gold nanorods (GNRs) were imaged by SPM-OCT and a home-made PS-OCT, respectively. A comparison of their imaging results proved that the SPM-OCT system can obtain the birefringence and dichroic parameters precisely and avoid the phase jump errors in the polarization image.
Besides, label-free interferometric imaging methods face challenges in distinguishing molecular signals from the background or the incoherent processes. To achieve molecular imaging, specific probes are typically required [28]. GNRs, as multifunctional probes, offer great potential for interferometric molecular imaging in oncology [29], ophthalmology [30], and respiratory medicine [31]. Intensity-based OCT molecular imaging requires a probe with different scattering intensities than the tissue [32,33]. Although studies have shown that GNRs can enhance the intensity of OCT [34], it is still difficult for intensity-based OCT to distinguish GNRs from biological tissues with high contrast. By comparing the polarization parameters of biological tissues and GNRs, it was found that the dichroic parameters obtained from our proposed method can distinguish GNRs from biological tissues.
2. METHODS AND MATERIALS
A. Principle of Polarization Parameter Calculation Based on Polarization Modulation
In this study, we proposed a polarization parameter calculation based on polarization modulation in interferometry. The schematic diagram of the interferometric system is shown in Fig. 1. Assume that the linear polarization direction of the light source is modulated in the
Figure 1.The system schematic diagram. PM Source, polarization-modulated light source; BS, beam splitter; RM, reflective mirror; Ref. Plane, reference plane.
The light output from the light source is divided into the reference arm and the sample arm through the beam splitter, respectively. The reference light returns directly to the beam splitter without changing the polarization state, whose electric field (
The returned sample light and the reference light interfere with each other at the beam splitter. The interference terms
The interference signal (
On the one hand, assume that the sample at the optical length difference
Based on Eqs. (1)–(6), the interference signal of the birefringent sample is proportional to the following formula:
By performing an inverse Fourier transform on the interference signal in the
Then, the
Based on Eqs. (8) and (9), the amplitudes of
Besides, the phase of
The retardation
On the other hand, assume that the sample is dichroic and the Jones matrix of the sample
Based on Eqs. (1)–(5) and (15), the interference signal of a dichroic sample is proportional to the following formula:
By performing an inverse Fourier transform on the interference signal in the
Then, the
Based on Eqs. (17) and (18), the amplitudes of
Besides, the phase of
We defined the diattenuation ratio
It can be seen that when the modulation frequency of the source is
B. Theory of the SPM Method
In this study, we proposed an SPM method that can obtain a complete polarization modulation signal in the spectral domain by a single acquisition. The SPM module is shown in Fig. 2. It includes the vertical linear polarizer (LP), the multi-order wave plate (MP) with the fast axis direction being
Figure 2.The schematic diagram of the SPM module.
The light passing the polarizer is in a vertical linear polarization state. Its Jones vector (
The wave plate can be considered as a birefringent device. Based on Eq. (6), the Jones matrix of the MP with the fast axis direction being
Besides, the Jones matrix of the AP with the fast axis direction being 0 rad to the vertical linear polarizer direction can be expressed as
Therefore, the Jones vector of the output light from the SPM module (
It can be seen that the Jones vector of the output light is the same as the expression of the polarization-modulated light source required for interferometric imaging with polarization modulation [Eq. (1)]. To avoid potential dimensional misunderstandings, we used
C. System Setup of SPM-OCT
The schematic diagram of the SPM-OCT system is shown in Fig. 3. The source light is emitted from a super luminescent diode (SLD) (BLM2-D, Superlum) with a center wavelength of 840 nm and a 3 dB bandwidth of 100 nm. Then, the light passes through a three-paddle polarization controller (PC) and is coupled to the SPM module. The SPM module includes a vertical linear polarizer (LP), two multi-order wave plates (MPs) (WPMH05M-830, Thorlabs) with the fast axis direction being 45 deg to the LP direction, and a vertical achromatic quarter wave plate (AP) (AQWP05M-980, Thorlabs). The purpose of using two MPs is to generate a suitable polarization modulation frequency that is easier to distinguish.
Figure 3.The schematic diagram of the SPM-OCT system. SLD, super luminescent diode; PC, polarization controller; FC, fiber collimator; LP, linear polarizer; MP, multi-order wave plates; AP, achromatic quarter wave plate; BS, non-polarizing beam splitter; DC, dispersion compensator; ND, neutral density filter; RM, reference mirror; GV, galvanometer scanning mirror; SL, scanning lens; SP, spectrometer.
The output light of the SPM module is separated into the sample and reference arms by a 50:50 non-polarizing beam splitter (BS). The reference light passes a dispersion compensator (DC) and a neutral density filter (ND) and is reflected by a reference mirror (RM). In the sample arm, after backscattering from the sample through the galvanometer scanning mirror (GV) and the scanning lens (SL), the sample light returns to the BS and interferes with the reference light. Then, the interference light is acquired by the commercial spectrometer (SP) (Cobra-S 800, Wasatch Photonics). The optical fiber used to connect the spectrometer to the collimator is a non-polarization-maintaining, low-birefringence optical fiber. Our system is performed with an axial resolution of
Besides, to verify the imaging capability of the SPM-OCT system, we also used the imaging result from a home-made single-input PS-OCT system as a reference. The accuracy of this PS-OCT system has been verified [35], which has been applied in many biological studies [36–38].
D. Imaging Algorithm of SPM-OCT
The flowchart of the SPM-OCT imaging algorithm is shown in Fig. 4(a). The specific steps are as follows.
Figure 4.(a) is the flowchart of the SPM-OCT imaging algorithm. (b) is the schematic diagram of the split-spectrum method.
Step 1: Acquire the spectral signal
Step 2.1: Perform a one-dimensional (1D) inverse Fourier transform (
Step 2.2: The intensity information
Step 3.1: Split the spectral signal
The premise for our proposed method is to distinguish the signal of the modulation frequency corresponding to the birefringence parameter (
Step 3.2: Calculate the modulation signals of birefringence (
Step 3.3: Calculate the retardation
The reason why the SPM-OCT system does not have phase jumps can be deduced theoretically. For a typical single-input PS-OCT, the interference signals of two polarization channels (
The optic axis is calculated from the phase of
It can be seen from Eq. (36) that the calculated phase (
Compared with the typical single-input PS-OCT, the SPM-OCT calculates the optic axis based on the Eq. (8). The coefficient of the
E. Calculation of the Contrast-to-Noise Ratio
The contrast-to-noise ratio (CNR) is used to evaluate the imaging contrast between the biological tissue and GNR. The CNR is defined as follows [40]:
F. Sample Preparation
The BALB/c nude mouse was used for the imaging experiment. After the mouse was sacrificed by anesthetic overdose, the skin of its leg was removed, and the skinned leg was imaged by PS-OCT and SPM-OCT, respectively.
To make a GNR phantom, the GNR was dispersed in the agar solution, then the agar solution was solidified to form a GNR phantom, and the phantom was imaged using PS-OCT and SPM-OCT, respectively. All experimental protocols using animals were approved by the Institutional Animal Care Committee of Nankai University.
3. RESULTS
A. Validation of SPM-OCT Measurements
To evaluate the accuracy of polarization measurements of the SPM-OCT system, a standard quarter-wave plate (QWP) (AQWP05M-980, Thorlabs) and a linear polarizer (LP) (LPVIS050-MP2, Thorlabs) were scanned by the SPM-OCT system, respectively. The QWP was used to verify the accuracy of birefringence parameters, including retardation (
The modulation signals obtained in the
Figure 5.(a1) and (a2) are the modulation signals obtained in the
By calculating the modulation signals obtained by measuring QWP and LP, the birefringence and dichroic parameters are obtained. A comparison of measured and theoretical values of birefringence parameters, including retardation (
As described in our previous study [35], the standard deviation of the retardation measured by PS-OCT is 4.31°. Besides, the optic axis values measured by PS-OCT slightly deviate from the ground truth. Comparing the validation results of SPM-OCT and PS-OCT, it can be seen that SPM-OCT has a lower retardation error than PS-OCT. The factors of SPM-OCT measurement errors include the retardation error of the achromatic quarter wave plate (AP) and the linear relationship between the wave numbers and the retardation of multi-order wave plates (MPs).
Besides, a comparison of measured and theoretical values of dichroic parameters, including the diattenuation ratio (
Figures 5(d1)–5(d4) are the retardation, optic axis, diattenuation ratio, and transmission axis B-scan images of the measured polarization elements at several angles. Angles shown in Figs. 5(d1) and 5(d2) correspond to the fast axis of the QWP. Angles shown in Figs. 5(d3) and 5(d4) correspond to the transmission axis of the LP. The QWP is constructed by aligning the fast axis of a birefringent plate with the slow axis of another birefringent plate. The LP consists of ellipsoid nanoparticles embedded in glass. The sample light enters the element and focuses on its lower surface [the horizontal line shown in Fig. 5(d)], whose polarization information is used to verify the polarization measurement capability of the system. The other interfaces of the polarization element are far from the focal plane, which are almost invisible due to their weak intensity. The different depth positions of the QWP and LP are because we adjusted the optical path length of the reference arm to accommodate the different thicknesses of these elements.
In summary, the consistency between the polarization measurement results and theoretical values demonstrates the high precision and reliability of the SPM-OCT system in polarization measurement.
B. Polarization Imaging Capability of SPM-OCT in Biological Tissues
The skinned mouse leg includes low-birefringence (lipid) and high-birefringence (muscle) tissues, making it an ideal biological tissue for evaluating the polarization measurement capability of the imaging system. To analyze the polarization measurement capability of SPM-OCT in biological tissues, SPM-OCT and PS-OCT were used to image the skinned mouse leg. Figures 6(a)–6(c) are the intensity, intensity-retardation, and intensity-optic axis B-scan images of the skinned mouse leg, respectively. Labels 1 and 2 represent the images obtained by PS-OCT and SPM-OCT, respectively. Among them, intensity-retardation and intensity-optic axis images are two-parameter merged images, which use intensity information to determine its brightness and polarization information to determine its hue. The optic axis ranges of SPM-OCT and PS-OCT are
Figure 6.(a)–(c) are the intensity, intensity-retardation, and intensity-optic axis B-scan images of the skinned mouse leg, respectively. Labels 1 and 2 represent the images obtained by PS-OCT and SPM-OCT, respectively. Scale bars are 1 mm.
Comparing Figs. 6(a1) and 6(a2), it can be seen that the intensity images obtained by PS-OCT and SPM-OCT are similar, and the high resolution of these systems ensures the clear presentation of lipid texture (circled by the yellow dotted line). As shown in Figs. 6(b1) and 6(b2), the retardation images obtained by PS-OCT and SPM-OCT are also similar. Among them, the lipid tissue shows low retardation values due to its low birefringence, while the muscle tissue with a high birefringence shows the retardation fluctuating with depth [38].
It is important to note that the optic axis images obtained by single-input PS-OCT may display erroneous information known as the phase jump. For example, the direction of the anisotropic structure of muscle fibers determines the value of the optic axis. Due to the directional continuity of the muscle fibers, the optic axis of the normal muscle usually does not have drastic changes within the imaging range of OCT. However, in the optic axis image obtained by single-input PS-OCT, a part of the muscle will show erroneous numerical mutations. As shown in Figs. 6(b1) and 6(c1), the region corresponding to the retardation changes from 0 to
Due to its three-dimensional imaging capability, OCT can not only provide B-scan images that display the tomographic structure but also provide
Figure 7.(a)–(c) are the intensity, intensity-retardation, and intensity-optic axis
Then, we enlarged the fiber tissue shown in the
The comparison results in this section proved that SPM-OCT not only can obtain various polarization parameters compared to the available PS-OCT but also can avoid the phase jump errors in the optic axis image, which greatly reduces the misreading of polarization information.
C. Feasibility of Distinguishing Biological Tissues and GNRs by SPM-OCT
In this section, the SPM-OCT was used to image the skinned mouse leg and GNR phantom, respectively, and explored the feasibility of distinguishing biological tissues and GNRs based on SPM-OCT. Figures 8(a)–8(d) are the
Figure 8.(a)–(d) are the retardation, optic axis, diattenuation ratio, and transmission axis images of the sample, respectively. Labels 1 and 2 represent the skinned mouse leg and GNR phantom, respectively.
The skinned mouse leg image mainly shows two types of tissues: muscle and lipid. As shown in Fig. 8(a1), muscles could show high retardation values due to their high birefringence, while low birefringent lipids show low retardation values. The optic axis image [Fig. 8(b1)] reflects the direction of the birefringent sample. The diattenuation ratio image [Fig. 8(c1)] shows the magnitude of the dichroism. It can be seen that the diattenuation ratio value of muscle and lipid is small, which is consistent with the current understanding that biological tissues have low dichroism [6]. The transmission axis image [Fig. 8(d1)] shows the transmitting direction of the dichroism. It can be seen that although the dichroism of biological tissues is small, Fig. 8(d1) can still show the different transmission axes of biological tissue.
The images of the GNR phantom show several GNRs dispersed in the solid agar. As shown in Fig. 8(a2), GNRs with birefringence show a certain retardation. The optic axis image [Fig. 8(b2)] shows the birefringent direction of the GNR. The diattenuation ratio image [Fig. 8(c2)] shows that the GNR has higher dichroism than biological tissue. The transmission axis image shows the dichroic direction of the GNR. It can be seen that there are differences in the optic axis and the transmission axis. This is because the GNR is not completely dispersed into individual particles, and the optic axis and the transmission axis reflect different combined effects of multiple GNRs. Comparing the polarization images of biological tissues and GNRs, the most obvious difference is the huge difference in the diattenuation ratio values between them. This difference is expected to be a key parameter for distinguishing biological tissues and GNRs.
To evaluate the effect of SPM-OCT in distinguishing biological tissues and GNRs, we quantified and compared the retardation and diattenuation of muscle, lipid, and GNR. The quantified areas of muscle and lipid are selected by dotted lines in Figs. 8(a1) and 8(c1). The box plots of the quantification results are shown in Fig. 9. The distribution of the cross-coupling coefficients is shown in the boxplots (Fig. 9). The 25%–75% indicates the range from the first quartile (Q1) to the third quartile (Q3), which is represented by a square box. The difference between the Q3 and the Q1 is called the interquartile range (IQR). The 1.5IQR rule is a statistical method used for identifying outliers in a dataset. The 1.5IQR range indicates the range from
Figure 9.Box plots of retardation and diattenuation of lipid, muscle, and GNR.
It can be seen that the median retardation value of the lipid is low, while that of the muscle is high. The retardation of GNR is higher than that of the lipids but lower than that of the muscle. The CNR of GNR relative to lipid and muscle in the retardation image is 3 dB and 0 dB, respectively, which shows that it is difficult to identify GNR in lipid and muscle using the retardation image.
In contrast, the diattenuation ratio of GNR is higher than that of biological tissue. This result confirms that the diattenuation values of most biological tissues are extremely low [41] and that GNRs have high diattenuation [42]. Outliers with high values are associated with the aggregation of multiple GNRs. Suppose a multidimensional polarization vector describes the polarization properties of the GNR. When there are multiple GNRs in the detection spot, the polarization vector detected by the system is the superposition of the polarization vectors of multiple GNRs, resulting in a change in the obtained polarization information.
The CNR of GNR relative to lipid and muscle in the diattenuation image is 25 dB and 20 dB, respectively, which are significantly higher than that of some intensity-based OCT molecular imaging methods [32,33]. This result demonstrated that the dichroic parameters obtained from SPM-OCT can distinguish GNRs from biological tissues.
4. DISCUSSION
Integrating polarization measurement with interferometric imaging has shown great potential for generating three-dimensional structural and polarization data simultaneously. However, these integrations, represented by PS-OCT, still face two major challenges. First, there is a trade-off between the acquisition time and the diversity of polarization parameters. Second, the dual-detector configurations increase the complexity and cost of the system, while single-detector-based methods face constraints in acquisition speed and imaging range. So far, we have developed various single-input PS-OCT systems, effectively applied in biological tissue evaluation [35–37] and high-resolution dataset construction [43–45]. Building upon our extensive research, it is imperative to develop a compact and efficient interferometric polarization measurement method.
To address these problems, we conducted the following work in this study. (1) An efficient SPM method was proposed to extract birefringence and dichroism parameters by interferometric imaging, meeting the demand for rapid and multi-parameter polarization measurement. (2) The SPM method was successfully integrated into an OCT system, creating an advanced interferometric imaging system (SPM-OCT) capable of capturing multi-dimensional polarization parameters without sacrificing imaging speed or axial range. The improvement in imaging speed of SPM-OCT is achieved by reducing acquisition times. Compared with the multi-input PS-OCT system using time multiplexing, the acquisition speed of SPM-OCT is twice that of dual-input PS-OCT and three times that of triple-input PS-OCT. As a single-input system, SPM-OCT can obtain the birefringence and dichroism parameters through a single acquisition, achieving a better suppression effect on motion artifacts caused by breathing and heartbeat. (3) The imaging results demonstrated that the SPM-OCT system can provide both birefringence and dichroic parameters while mitigating phase jump errors, differentiating biological tissues from GNRs based on the dichroic parameter.
At present, most of the interferometric polarization modulation methods belong to time-sequenced mechanisms based on electro-optic modulator [18,24]. Since many interferometric imaging methods are wide-spectrum-based methods, we designed an SPM method to generate the polarization modulation signal in the wavenumber domain. The modulation signals can be obtained through a single acquisition, thus reducing the acquisition time. Besides, the system structure only changes the light source of a conventional interferometric imaging system. Therefore, the system design is very beneficial to endow existing systems with polarization detection functions, making it highly valuable for the product upgrading of interferometric imaging systems.
The SPM-OCT combines the OCT and the SPM method, which has the advantages of rapid acquisition, comprehensive polarization parameters, and simplified single-detector configuration. The imaging capability of SPM-OCT was verified by imaging standard polarization elements, biological tissues, and GNR phantoms, proving the high reliability, accuracy, and application value of the SPM-OCT system. It should be noted that structurally anisotropic nanomaterials, including gold nanorods, usually have significantly higher dichroism than biological tissues. The high sensitivity of SPM-OCT to dichroic materials makes it a potential method for achieving OCT molecular imaging.
Future research will proceed in the following directions. (1) Combine the SPM method with various interferometric imaging methods to endow and optimize their polarization detection function. (2) Incorporate the depolarization calculation methods from existing PS-OCT frameworks and design parameters to describe depolarization and local polarization obtained by SPM-OCT. (3) Refine both hardware components and signal processing techniques to further improve the resolution and accuracy of the system. (4) Investigate ways to expand the design and application of the SPM-OCT system in the field of medical diagnostics.
5. CONCLUSION
In this study, we developed the SPM method to perform simple and rapid interferometric polarization measurement and constructed the SPM-OCT system to extract birefringence and dichroic parameters without reducing the acquisition rate or the axial imaging range. Through imaging experiments on standard polarization elements, biological tissues, and GNR phantoms, we demonstrated that our method not only offers accurate birefringent and dichroic parameters but also effectively avoids phase jump errors. The ability of SPM-OCT to differentiate GNRs from biological tissues further underscores its potential for clinical and research applications. The efficient polarization measurement mechanism of the SPM method is anticipated to promote the development of interferometric imaging and inspire new research directions in polarization imaging technology.
References
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[11] S. Banerjee, S. Aoki, K. Kitamoto. Regional contrast comparison between OCT intensity and polarization-sensitive OCT images for myopic and macular defect suspects. Invest. Ophthalmol. Vis. Sci., 64, 1071(2023).

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