How to mark lesions

Abstract

Disclosed herein is a method for marking lesions. The disclosed method can assist surgeons in locating small nodules by marking the nodules with a reference coil immersed in a fluorescent dye. In some preferred embodiments, the reference coil is placed at the time of biopsy using a robotic surgical system. In alternative embodiments, non-robotic peripheral navigation platforms can also be effectively used as an adjunct to surgical resection. The dye marks the location of the nodule, making it visible and palpable during surgery. The surgeon can then more precisely target the cancerous tissue. The disclosed method allows the reference coil to be placed several days before surgery.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application Nos. 63 / 478,994, filed January 9, 2023; 63 / 369,228, filed July 23, 2022; and 63 / 369,129, filed July 22, 2022, which claims the benefit of U.S. Provisional Patent Application No. 18 / 173,891, filed February 24, 2023, the disclosures of which are incorporated herein by reference in their entireties. [Background technology]

[0002] Pulmonary nodules are now more easily identified due to the increasing use of computed tomography (CT) in chest and lung cancer screening efforts. The rate of early-stage lung cancer is also increasing. While surgical resection is the standard of care at this stage, interest in and growing literature supporting lung-sparing surgery for both new lung cancers and secondary pulmonary metastases necessitates accurate localization techniques to provide optimal results. An ideal localization procedure provides multiple options for surgeons to clearly identify these nodules and allow the best opportunity for preserving as much healthy lung tissue as possible.

[0003] However, surgical localization of pulmonary nodules can present multiple challenges. Several tumor factors, including nodule size, distance from the pleural surface, and nodule nature (ground glass, solid, or mixed), as well as factors such as associated structural lung disease, the presence of anthracite pigment on the lung surface, and pleural adhesions, have important implications for localization and surgical resection. Other potential barriers include access to treatment / procedure listings, institutional logistical challenges, and reliance on open surgical techniques.

[0004] Diagnosis and treatment of pulmonary nodules can be combined during surgical resection, but this approach can lead to the removal of benign lesions and excess healthy lung tissue if the lesion is difficult to localize. Preoperative bronchoscopically guided marking of pulmonary nodules can facilitate this process, but current technology is limited and often suffers from the problem of dye markings disappearing if surgery is performed several days later.

[0005] With the increasing interest in lung-sparing surgery for both de novo lung cancer and secondary pulmonary metastases, accurate localization is now more important to provide desirable outcomes. An ideal localization procedure provides surgeons with multiple options for accurately identifying pulmonary nodules.

[0006] Therefore, methods for accurate marking of lesions remain necessary for accurate identification of pulmonary nodules after a clinically significant period of time has passed. Summary of the Invention [Means for solving the problem]

[0007] This disclosure describes a novel method for addressing the problem of surgical localization of lung nodules, increasing the efficiency between diagnosis and subsequent surgery. The disclosed method can assist surgeons in locating small nodules by marking the nodule with a reference coil immersed in a fluorescent dye, such as indocyanine green (ICG). In some preferred embodiments, the reference coil is placed at the time of biopsy using a robotic bronchoscope system. In alternative embodiments, non-robotic peripheral navigation platforms can also be effectively used as an adjunct to surgical resection. The dye marks the location of the lung nodule, making it visible and palpable during surgery. Surgeons can then more precisely target cancerous tissue, enabling lung-sparing surgery. The disclosed method allows the reference coil to be placed several days before surgery.

[0008] Although the above description describes a method for marking lesions in the lungs, the disclosed method can be used to mark lesions in other tissues and organs in the body.

[0009] Additionally, while the above description describes methods of marking lesions using particular coils, it is contemplated that any biocompatible coil suitable for use in the human body can be used in the disclosed methods, including coils constructed from biocompatible metals or alloys, biocompatible polymers, or other biocompatible materials.

[0010] Additionally, although the above description describes the use of particular biocompatible fluorescent dyes, it is contemplated that any biocompatible fluorescent dye suitable for use in the human body can be used in the disclosed methods.

[0011] Also, while the above description describes the process of dipping the dye-dipped coil into the dye immediately prior to placing the coil as a marker, it is contemplated that the coil can instead be pre-dipped into an appropriate amount of fluorescent dye and that this pre-dipped coil be used in the disclosed method. The shelf life of the pre-dipped coil will vary depending on the particular dye and material comprising the coil.

[0012] Finally, while the above description describes the use of a coil that is fully immersed in dye, it is also contemplated that only a portion of the coil may be immersed in dye and that such a partially immersed coil may be used in the disclosed method. [Brief explanation of the drawings]

[0013] [Figure 1] FIG. 1 illustrates an embodiment of a dye immersion coil for use in the disclosed method. [Figure 2A-B] Figures 2A-2B show an embodiment of a dye-dipped coil delivered to a nodule site via a robotic catheter, with Figure 2A showing a cross-sectional CT image with a fiducial marker placed in the right upper lobe with a ground-glass lesion and Figure 2B showing a cross-sectional CT image with a fiducial marker placed in the right upper lobe with a solid nodule. [Figure 3]Figure 3 shows visualization of an embodiment of a dye-immersed coil as a fluorescent target on the pleural surface. [Figure 4A] FIG. 4A shows a diagram of an embodiment of a robotic pulmonary wedge resection performed successfully using light emission provided by a dye-immersion coil. [Figure 4B] FIG. 4B shows an embodiment of a wedge resection showing the reference coil in the center of the specimen. DETAILED DESCRIPTION OF THE INVENTION

[0014] This disclosure describes a novel method for addressing the problem of surgical localization of pulmonary nodules, increasing the efficiency between diagnosis and subsequent surgery. The disclosed method can assist surgeons in locating small nodules by marking the nodule with a reference coil immersed in a fluorescent dye, such as indocyanine green (ICG). In some preferred embodiments, the reference coil is placed at the time of biopsy using a robotic bronchoscope system. In alternative embodiments, non-robotic peripheral navigation platforms can also be effectively used as an adjunct to surgical resection. The dye marks the location of the pulmonary nodule, making it visible and palpable during surgery. Surgeons can then more precisely target the cancerous tissue, enabling lung-sparing surgery.

[0015] The disclosed method allows for the placement of a reference coil several days before surgery.

[0016] In one example study conducted at multiple sites, an ICG dye-soaked coil was placed in patients' biopsy-confirmed cancerous lung nodules in preparation for surgery several days later. The coil placement occurred 0–9 days before surgery. Because the dye was present in the coil, it remained fluorescent and visible as a bright neon green for at least 5 days after surgery. The dye's bright green light allowed each thoracic surgeon to precisely locate the area to be resected, minimizing the amount of lung tissue removed and resulting in a better outcome for the patient. Furthermore, the coil is palpable and visible via intraoperative fluoroscopy or ultrasound, allowing surgeons multiple options for locating the lesion based on their experience and expertise. When available, imaging techniques such as cone-beam CT or magnified fluoroscopic imaging can also be used to assist in assessing the coil's position relative to the nodule, thereby supporting accurate resection.

[0017] The extended time allowed between use of the disclosed method of placing markers and subsequent surgery provides a significant advantage to patients in small, rural, regional hospitals where the availability of on-site thoracic surgery is limited. Patients may require time to schedule, prepare for, and travel to larger medical centers for surgery. The disclosed method of fiducial marking means that when a patient arrives at the surgery center to undergo surgery, proper localization and visualization are possible to assist the surgeon in the resection.

[0018] The disclosed methods benefit patients by providing surgeons with more efficient, faster, and accurate surgical procedures that better preserve healthy tissue. The disclosed methods substantially improve the success of lung-sparing surgery for both primary and secondary malignancies of the lung.

[0019] Although the above description describes a method for marking lesions in the lungs, the disclosed method can also be used to mark lesions in other tissues and organs in the body.

[0020] Additionally, while the above description describes methods of marking lesions using particular coils, it is contemplated that any biocompatible coil suitable for use in the human body can be used in the disclosed methods, including coils constructed from biocompatible metals or alloys, biocompatible polymers, or other biocompatible materials.

[0021] Additionally, although the above description describes the use of particular biocompatible fluorescent dyes, it is contemplated that any biocompatible fluorescent dye suitable for use in the human body can be used in the disclosed methods.

[0022] Also, while the above description describes the process of dipping the dye-dipped coil into the dye immediately prior to placing the coil as a marker, it is contemplated that the coil can instead be pre-dipped into an appropriate amount of fluorescent dye and that this pre-dipped coil be used in the disclosed method. The shelf life of the pre-dipped coil will vary depending on the particular dye and material comprising the coil.

[0023] Finally, although the above description describes the use of a coil that is fully immersed in dye, it is also contemplated that only a portion of the coil may be immersed in dye and that such a partially immersed coil may be used in the disclosed method. [Example]

[0024] The following examples are provided as specific illustrations of the disclosed methods, however, it should be understood that the invention is not limited to the specific details set forth in the examples.

[0025] Additionally, any range of numerical values recited above or in the following paragraphs describing or claiming various aspects of the invention, e.g., ranges expressing a particular set of properties, units of measure, conditions, physical states, or percentages, is intended to expressly and literally incorporate herein by reference or otherwise any numerical values falling within such ranges, including any numerical values or any subset of ranges encompassed within any such recited range. The term "about," when used as a modifier of or in connection with a variable, is intended to convey that the numerical values and ranges disclosed herein may be flexible as understood by one of ordinary skill in the art, and that practice by one of ordinary skill in the art of the disclosed invention using temperatures, concentrations, amounts, contents, and properties outside the literal ranges will achieve desired results.

[0026] Case studies The following case studies illustrate the disclosed methods.

[0027] Materials and Methods One patient from El Camino Hospital / Palo Alto Medical Foundation in Mountain View, CA, and three patients from Aurora Medical Center in Kenosha, Kenosha, WI, with incidentally detected pulmonary nodules were selected for a proof-of-concept study of the disclosed method. The selection was based on the scheduled time between the marking procedure and thoracic surgery (0, 4, 5, and 9 days between placement of the ICG dye-immersed coil and surgical resection).

[0028] The material used in the procedure was a fiducial coil (G10417 Tornado Embolization Coil 7 mm × 3 mm × 0.035 in × 8 cm, Cook Medical, Bloomington, IN, USA) immersed in indocyanine green dye (ICG) (NDC-70100-424-02, HUB Pharmaceuticals, Scottsdale, AZ, USA) for 10 minutes before being replaced by robotic bronchoscopy using the Ion Endoluminal System (Intuitive Surgical, Sunnyvale, CA, USA) 0–9 days before thoracic surgery.

[0029] All patients underwent robot-assisted navigational bronchoscopy (RANB) using the Ion Endoluminal System (Intuitive Surgical, Sunnyvale, CA, USA) with cone-beam computed tomography or 3D fluoroscopy used as an additional tool for secondary confirmation. El Camino Hospital used a ceiling-mounted Artis Zee cone-beam scanner (Siemens Medical Solutions, Malvern, PA, USA) (n = 1), and Aurora Medical Center in Kenosha used a mobile C-arm Cios Spin (Siemens Medical Solutions, Malvern, PA, USA) (n = 3). All patients had incidentally detected new nodules (two solid nodules and two ground-glass opacity nodules). None of the patients had a history of allergic reactions to contrast agents. Treatment was planned as diagnostic bronchoscopy with biopsy, followed by marking (if intraprocedural cytopathology showed atypical cells or confirmed malignancy), and then surgical resection (scheduled for a later date at the Kenosha, WI facility or the same day at the Mountain View, CA facility).

[0030] This procedure was performed using robotic-assisted navigation bronchoscopy by marking the lesion with an ICG dye-soaked Cook Tornado Coil, as shown in Figure 1. Using a standard 1 ml tuberculin syringe with a Luer lock, 0.25 ml of a diluted mixture of ICG (25 mg of ICG dye mixed with 10 ml of sterile water to achieve an ICG dye concentration of 2.5 mg / ml) was aspirated and injected into the Cook Tornado coil cartridge sheath, priming the sheath and placing the coil within it, immersing it in the ICG dye. A 10-minute waiting period was required for the dye to completely immerse the coil. This time was selected to allow sufficient soaking of the synthetic fibers of the coil. After the waiting period, an additional 0.25 ml of the same ICG dye mixture was injected into the coil sheath for additional priming, just before placing the ICG dye-soaked coil into the target nodule. The ICG-impregnated coil was then retrofitted onto the superDimension delivery catheter and placed by pushing the guidewire provided in the catheter kit (superDimension™ Marker Delivery Kit, Medtronic, MN, USA).

[0031] An ICG immersion coil was placed within the target lesion as shown in Figure 2 , and then, on the same day as a single anesthesia procedure at Mountain View, one patient was taken directly to the operating room for a robotic wedge resection (da Vinci, Intuitive Surgical, Sunnyvale, CA, USA) and three patients from Kenosha were taken to the recovery room and then discharged. These patients underwent robotic thoracic surgery at another Aurora Medical Center facility with available thoracic surgery services 4, 5, and 9 days later, respectively.

[0032] result Four patients underwent treatment according to embodiments of the disclosed methods described herein. All lesions were <2 cm on pre-treatment chest CT (two ground-glass nodules measuring 12 mm and 15 mm and two solid nodules measuring 10 mm and 15 mm), located in the outer third of the lung, and without bronchial manifestations.

[0033] ICG dye-immersed coils were placed on biopsy-confirmed cancerous lung nodules in patients in preparation for surgery, both immediately after the procedure (day 0) and several days later (days 4, 5, and 9). Using the Firefly Fluorescence Imaging Vision System (da Vinci Fluorescence Imaging Vision System, da Vinci, Intuitive Surgical, Sunnyvale, CA, USA) on a da Vinci robotic system (da Vinci, Intuitive Surgical, Sunnyvale, CA, USA), localization of the nodules was successful in all cases by visualizing the dye-immersed coil as a neon-green target on the pleural surface, as shown in Figure 3. Using the neon-green illumination provided by the dye-immersed coil, localization and resection were precise, and margin-negative wedge resections were successfully performed in all four cases, as shown in Figures 4A-4B. Patients who met the criteria for further anatomical lung resection based on the frozen pathology results of the wedge resections then underwent further resections during that same anesthesia episode. No intraoperative or postoperative complications were observed. Because the coil is made of platinum and spaced synthetic fiber, the ICG dye adheres to the coil, remaining fluorescent and visible as a bright neon green for at least nine days. The bright green dye emission allows thoracic surgeons to precisely identify the resection site, potentially reducing the amount of lung tissue removed. Previously, it was impossible to localize nodules nine days after ICG dye marking without the coil because the dye effect fades within a few hours due to its metabolism and half-life. The ICG dye-soaked coil marks the nodule location, making it visible under the da Vinci robotic system's Firefly Fluorescence imaging system, allowing surgeons to more precisely target lesions and enable lung-sparing surgery. The coil is also palpable with surgical grasping forceps and visible intraoperatively under fluoroscopy or ultrasound, giving surgeons multiple options for using the chemical and mechanical properties of the dye-impregnated coil to localize lesions, depending on their experience and expertise.

[0034] Analysis of exemplary treatments Localization of nodules facilitates surgical resection and prevents the removal of benign disease and healthy lung tissue. However, successful localization has no clear definition and is usually subjectively assessed by the proceduralist. Failure can occur if the surgeon is unable to clearly visualize or palpate the lesion, leading to increased procedure time, resection of healthy lung tissue, changes in the surgical approach, and possible failure to remove the intended target. The failure to detect pulmonary nodules greater than 5 mm from the pleural surface and smaller than 10 mm in size exceeds 60% during video-assisted thoracoscopic approaches. In addition, pure ground-glass lesions are difficult to palpate. In the exemplary patient described, all lesions were <15 mm, located in the outer third of the lung, and showed no signs of bronchial tubercle. Two lesions were solid and two were ground-glass lesions. No dye dispersion was observed up to 9 days after marking, and all lesions were localized intraoperatively.

[0035] The ICG-immersed reference coils were used because they are made of platinum and porous synthetic fibers, allowing ICG dye to adhere to them. The deployed coil configuration, with a wide base and tapered design, is suitable for placement in small airways and lung parenchyma. The deployed coil is also easily felt with fingers or grasping forceps. The deployed configuration of the dye-impregnated coils is well visualized by fluoroscopy, ultrasound, or illumination. This design allows the fluorescent, bright neon green dye to remain visible for many days, as confirmed in a model patient at least 9 days later. The bright green dye emission allows thoracic surgeons to accurately identify the resection site, potentially reducing the amount of lung tissue removed. Previously, it was impossible to locate nodules 9 days after ICG dye marking without the use of coils because the dye effect fades within a few hours due to its metabolism and half-life. The ICG dye-impregnated coil marks nodule location and is visible under the da Vinci robotic system's Firefly Fluorescence imaging system, allowing surgeons to more precisely target lesions and enable lung-sparing surgery. The coil is also palpable with surgical graspers and visible under intraoperative fluoroscopy and ultrasound, giving surgeons multiple options for using the dye-impregnated coil's chemical and mechanical properties to localize lesions, depending on their experience and expertise.

[0036] We hypothesize that dye retention after several days is due to interactions between the ICG dye and the synthetic fibers of the coil. ICG has a short half-life upon injection, is prone to nonspecific protein binding, and is prone to poor photostability; however, these limitations can be overcome by conjugating ICG to other substances, such as lipids, metals, or nanoparticles. Consequently, we aimed to conjugate ICG to the synthetic fibers of the coil to alleviate these limitations while preserving its beneficial properties, namely, enhanced visualization.

[0037] While dye-immersed coils were placed in biopsies of confirmed or suspected malignant lesions in exemplary patients, the disclosed method can also be used for non-diagnostic lesions where suspicion is high and surgical biopsy or resection is planned. The additional time allowed before surgery with this ICG fiducial marker technique may provide significant benefits to patients in small rural or underserved hospitals where in-house thoracic surgery is scarce or limited. Similarly, larger academic hospitals where a single anesthesia procedure for localization followed by immediate surgery may be difficult to plan due to scheduling conflicts or procedural logistics may also benefit from this approach. Patients may also require time to schedule, prepare, and travel to larger medical centers.

[0038] We demonstrate that when robot-assisted nodule marking is performed 0 to 9 days after surgical resection, the intensity of neon-green emission from the ICG dye-immersed coil remains constant rather than fading or dispersing over time at the pleural surface. This approach facilitates patient access to later surgical procedures and allows for proper localization and visualization to assist the surgeon in resection. This may enhance the ability to successfully perform lung-sparing surgery for both primary and secondary lung malignancies.

[0039] In vitro experiments In vitro assessment of ICG retention with fiducial coil markers was performed.

[0040] In an exemplary experiment, a 25 mg ICG vial was filled with 10 cm of ICG, the same dilution utilized in vivo. 3 The diluted ICG solution was reconstituted with 100 ml of sterile water. Two types of coils were selected for the experiment: a Cook Medical Tornado 7 mm x 3 mm coil with nylon fibers and a Boston Scientific Vortex 182 mm x 3 mm coil with polyester fibers. The diluted ICG solution was slowly injected into the coil cartridge, creating a 0.5 cm 3 The coil was then soaked in diluted ICG. The ICG infusion coil was then left in place for 10 minutes. An additional 0.5 cm was added before implantation into the tissue.3 of diluted ICG was flowed into the cartridge sheath of the coil.

[0041] Freshly harvested, refrigerated beef liver was used in the experiment. The whole organ was cut into large pieces (approximately 10 cm x 5 cm). Two small incisions, at least 5 cm apart, were made in the liver tissue with a blade. Each incision was approximately 2 cm deep, as confirmed by a tape measure. One Tornado 7 x 3 mm ICG-impregnated coil and one Vortex 2 x 3 mm ICG-impregnated coil were then inserted into the tissue with forceps to a depth of 2 cm, at least 5 cm apart. The incisions were then sutured with 2-0 silk and tape was applied over them to mark the incision sites. Five such pieces were formed. These pieces were then individually packed in airtight plastic bags and placed in a container. The container was then placed in a freezer.

[0042] Seventy-seven days after the initial implantation, the cooled tissue was removed from the freezer. The tissue was removed from the container and allowed to thaw at room temperature for approximately 60 minutes. The tape and sutures were removed from the surface of the tissue. The incision site remained sealed in all specimens, and no coils were visible on visual inspection. The room in which the results were analyzed was darkened, and then each segment of tissue was individually illuminated using the firefly light on the Intuitive Surgical Xi robot. All coils glowed and were easily visible.

[0043] conclusion A novel method is described for addressing the problem of surgical localization of lesions or nodules in tissues or organs. The disclosed method increases efficiency between diagnosis and subsequent surgery. The disclosed method can assist surgeons in localizing small nodules by marking the nodules with a reference coil immersed in fluorescent dye. The reference coil can be used, for example, to mark lung nodules and can be placed during biopsy using, for example, a robotic bronchoscopy system. Alternatively, non-robotic peripheral navigation platforms can also be effectively used as an adjunct to surgical resection. The dye marks the location of the nodule, making it visible and palpable during surgery. The surgeon can then more precisely target the cancerous tissue. The disclosed method allows the reference coil to be placed several days before surgery.

[0044] While the above examples illustrate methods for marking lesions in the lung or liver, the disclosed methods can be used to mark lesions in other tissues and organs within the body. Additionally, while the examples demonstrate the use of robot-assisted navigational bronchoscopy (RANB) for placement of dye-labeled reference coils in lung tissue, it is contemplated that other techniques can be used for placement of dye-labeled reference coils in lung tissue or other organs or tissues, including, but not limited to, image-guided endoscopic techniques using an endoscope, catheters, or needles or catheters, and image-guided percutaneous techniques using needles or catheters. Such techniques include, for example, endoscopic ultrasound and CT-guided needle placement. Furthermore, while the examples illustrate methods for marking lesions using specific coils, it is contemplated that any biocompatible coil suitable for use in the human body can be used with the disclosed methods, including coils constructed from biocompatible metals or alloys, biocompatible polymers, or other biocompatible materials. Furthermore, while the examples illustrate the use of specific biocompatible fluorescent dyes, it is contemplated that any biocompatible fluorescent dye suitable for use in the human body can be used with the disclosed methods. Also, while the examples illustrate a procedure in which the dye-dipped coil is dipped in the dye immediately prior to placement as a marker, the coil can alternatively be pre-dipped in an appropriate amount of fluorescent dye, and it is contemplated that this pre-dipped coil will be used in the disclosed methods. The shelf life of the pre-dipped coil will vary depending on the particular dye and materials comprising the coil. Finally, while the examples illustrate the use of a coil that is completely dipped in the dye, it is also contemplated that only a portion of the coil is dipped in the dye, and that such a partially dipped coil will be used in the disclosed methods.

[0045] The foregoing description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the invention(s) disclosed herein. While various inventive aspects have been disclosed in the context of specific embodiments, implementations, and examples that have been illustrated, those skilled in the art will recognize that the invention extends beyond the specifically disclosed embodiments to other alternative embodiments and / or uses of the invention, as well as obvious modifications and equivalents thereof. In addition, while several variations of various inventive aspects have been shown and described in detail, other variations within their scope will be readily apparent to those skilled in the art upon review of this disclosure. It should also be understood that the scope of the disclosure includes various combinations or subcombinations of the specific features and aspects of the embodiments disclosed herein, and therefore various features, modes of implementation, and aspects of the disclosed subject matter can be combined with or substituted for one another. Generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the disclosure. Thus, the present disclosure is not intended to be limited to the embodiments shown herein, but is intended to be accorded the widest scope consistent with the principles and novel features disclosed herein.

[0046] All references cited herein are expressly incorporated herein by reference.

Claims

1. 1. A method for marking a lesion, comprising: a. immersing a reference coil in a solution containing a fluorescent dye to create a dye-immersed reference coil; and b. placing the dye-immersed reference coil over or within a target lesion or nodule in mammalian tissue. in order, marking the lesion using the dye-immersed reference coil configuration; The method, wherein after the dye-immersion reference coil is placed on or within the lesion, the dye-immersion reference coil is visible using an imaging technique.

2. 10. The method of claim 1, wherein the dye-immersed reference coil is positioned over the target lesion or nodule using an image-guided endoscopic technique using an endoscope, needle, or catheter, or an image-guided percutaneous technique using a needle or catheter.

3. 10. The method of claim 1, wherein the dye-immersed reference coil is positioned over the target lesion or nodule using a robot-assisted navigational bronchoscopy (RANB) technique.

4. The method of claim 1 , wherein the dye is indocyanine green.

5. The method of claim 2 wherein the dye is indocyanine green.

6. The method of claim 3 , wherein the dye is indocyanine green.

7. The method of claim 1 , wherein the mammalian tissue is human tissue.

8. 8. The method of claim 7, wherein the human tissue is human lung tissue.

9. 9. The method of claim 8, wherein the dye-immersed reference coil is positioned over the target lesion or nodule using a robotic-assisted navigational bronchoscopy (RANB) technique.

10. 10. The method of claim 9, wherein the dye is indocyanine green.

11. 1. A method of marking a lesion, comprising placing a dye-immersed reference coil over or within a target lesion or nodule in mammalian tissue, marking the lesion or nodule using the dye-immersed reference coil configuration; After placing the dye-immersion reference coil on or within the lesion or nodule, the dye-immersion reference coil can be identified using an imaging technique.

12. 12. The method of claim 11, wherein the dye-immersed reference coil is positioned over the target lesion or nodule using an image-guided endoscopic technique using an endoscope, needle, or catheter, or an image-guided percutaneous technique using a needle or catheter.

13. 12. The method of claim 11, wherein the dye-immersed reference coil is positioned over the target lesion or nodule using a robotic-assisted navigational bronchoscopy (RANB) technique.

14. The method of claim 11 , wherein the dye is indocyanine green.

15. The method of claim 12, wherein the dye is indocyanine green.

16. 14. The method of claim 13, wherein the dye is indocyanine green.

17. 12. The method of claim 11, wherein the mammalian tissue is human tissue.

18. 18. The method of claim 17, wherein the human tissue is human lung tissue.

19. 20. The method of claim 18, wherein the dye-immersed reference coil is positioned over the target lesion or nodule using a robotic-assisted navigational bronchoscopy (RANB) technique.

20. 20. The method of claim 19, wherein the dye is indocyanine green.

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