Facilitating clinically relevant skin tumor diagnostics with spectroscopy-driven machine learning.

In the dawning era of artificial intelligence (AI), health care stands to undergo a significant transformation with the increasing digitalization of patient data. Digital imaging, in particular, will serve as an important platform for AI to aid decision making and diagnostics. A growing number of studies demonstrate the potential of automatic pre-surgical skin tumor delineation, which could have tremendous impact on clinical practice. However, current methods rely on having ground truth images in which tumor borders are already identified, which is not clinically possible. We report a novel approach where hyperspectral images provide spectra from small regions representing healthy tissue and tumor, which are used to generate prediction maps using artificial neural networks (ANNs), after which a segmentation algorithm automatically identifies the tumor borders. This circumvents the need for ground truth images, since an ANN model is trained with data from each individual patient, representing a more clinically relevant approach.

Photoacoustic imaging of periorbital skin cancer ex vivo: unique spectral signatures of malignant melanoma, basal, and squamous cell carcinoma.

Radical excision of periorbital skin tumors is difficult without sacrificing excessive healthy tissue. Photoacoustic (PA) imaging is an emerging non-invasive biomedical imagi--ng modality that has potential for intraoperative micrographic control of surgical margins. This is the first study to assess the feasibility of PA imaging for the detection of periocular skin cancer. Eleven patients underwent surgical excision of periocular skin cancer, one of which was a malignant melanoma (MM), eight were basal cell carcinomas (BCCs), and two squamous cell carcinomas (SCCs). Six tumors were located in the eyelid, and five in periocular skin. The excised samples, as well as healthy eyelid samples, were scanned with PA imaging postoperatively, using 59 wavelengths in the range 680-970 nm, to generate 3D multispectral images. Spectral unmixing was performed using endmember spectra for oxygenated and deoxygenated Hb, melanin, and collagen, to iden--tify the chromophore composition of tumors and healthy eyelid tissue. After PA scanning, the tumor samples were examined histopathologically using standard hematoxylin and eosin staining. The PA spectra of healthy eyelid tissue were dominated by melanin in the skin, oxygenated and deoxygenated hemoglobin in the orbicularis oculi muscle, and collagen in the tarsal plate. Multiwavelength 3D scanning provided spectral information on the three tumor types. The spectrum from the MM was primarily reconstructed by the endmember melanin, while the SCCs showed contributions primarily from melanin, but also HbR and collagen. BCCs showed contributions from all four endmembers with a predominance of HbO2 and HbR. PA imaging may be used to distinguish different kinds of periocular skin tumors, paving the way for future intraoperative micrographic control.

Laser Speckle Contrast Imaging of the Blood Perfusion in Glabellar Flaps Used to Repair Medial Canthal Defects.

BACKGROUND: The glabellar flap is a common technique for surgical repair after tumor excision in the medial canthal area. However, the outcome may be affected by partial flap necrosis. Little is known about the impact of surgery on blood perfusion and the postoperative course of reperfusion due to the absence of reliable and noninvasive perfusion monitoring techniques. The aim of this study was to use a modern imaging technique to assess blood perfusion in glabellar flaps. METHODS: Glabellar flaps were used to repair medial canthal defects following tumor excision in 7 patients. Blood perfusion was monitored using laser speckle contrast imaging: during surgery, immediately postoperatively (0 weeks), and at follow-up, 1, 3, and 6 weeks after surgery. RESULTS: Perfusion decreased gradually along the length of the flap, and reached a minimum 15 mm from the flap base. Perfusion in the proximal 20 mm of the flap was completely restored after 1 week, while the distal part of the flap was gradually reperfused over 6 weeks. Both the functional and esthetic surgical outcomes were excellent. CONCLUSIONS: The rapid reperfusion of the glabellar flap may be explained by its connection to the vascular network via the flap pedicle. In flaps longer than 20 mm, the distal part can be considered a free skin transplant, and a combination of a glabellar flap and a free skin graft could then be considered.

Blood Perfusion of Human Upper Eyelid Skin Flaps Is Better in Myocutaneous than in Cutaneous Flaps.

BACKGROUND: The aim of this study was to monitor blood perfusion in human upper eyelid skin flaps and examine how the perfusion is affected by the thickness of the flap. METHODS: Twenty upper eyelids were dissected as part of a blepharoplasty procedure in patients. The medial end of the blepharoplasty flap remained attached to mimic a flap design often used in reconstruction in the periocular area, a myocutaneous flap in which the blood supply follows the fibers of the orbicularis muscle and is thus parallel to the long axis of the flap. The muscle was thereafter dissected from the flap to create a cutaneous flap. Blood perfusion in the 2 types of flaps was compared using laser speckle contrast imaging. RESULTS: Blood perfusion decreased gradually from the base to the tip of all the flaps. Perfusion was significantly higher in the myocutaneous flaps than in the cutaneous flaps (p < 0.0004): 69% in the myocutaneous flaps and 43% in the cutaneous flaps, measured 5 mm from the base. Blood perfusion was preserved to a greater extent distally in the myocutaneous flaps (minimum value seen at 25 mm) than in the cutaneous flaps (minimum seen at 11 mm). CONCLUSIONS: Blood perfusion was better preserved in myocutaneous flaps, including both skin and the orbicularis oculi muscle, than in cutaneous flaps. This may be of clinical interest in patients with poor microcirculation in which a long flap is required for reconstructive surgery.

Automatic threshold selection algorithm to distinguish a tissue chromophore from the background in photoacoustic imaging.

The adaptive matched filter (AMF) is a method widely used in spectral unmixing to classify different tissue chromophores in photoacoustic images. However, a threshold needs to be applied to the AMF detection image to distinguish the desired tissue chromophores from the background. In this study, we propose an automatic threshold selection (ATS) algorithm capable of differentiating a target from the background, based on the features of the AMF detection image. The mean difference between the estimated thickness, using the ATS algorithm, and the known values was 0.17 SD (0.24) mm for the phantom inclusions and -0.05 SD (0.21) mm for the tissue samples of malignant melanoma. The evaluation shows that the thickness and the width of the phantom inclusions and the tumors can be estimated using AMF in an automatic way after applying the ATS algorithm.

Comparison of photoacoustic imaging and histopathological examination in determining the dimensions of 52 human melanomas and nevi ex vivo.

Surgical excision followed by histopathological examination is the gold standard for the diagnosis and staging of melanoma. Reoperations and unnecessary removal of healthy tissue could be reduced if non-invasive imaging techniques were available for presurgical tumor delineation. However, no technique has gained widespread clinical use to date due to shallow imaging depth or the absence of functional imaging capability. Photoacoustic (PA) imaging is a novel technology that combines the strengths of optical and ultrasound imaging to reveal the molecular composition of tissue at high resolution. Encouraging results have been obtained from previous animal and human studies on melanoma, but there is still a lack of clinical data. This is the largest study of its kind to date, including 52 melanomas and nevi. 3D multiwavelength PA scanning was performed ex vivo, using 59 excitation wavelengths from 680 nm to 970 nm. Spectral unmixing over this broad wavelength range, accounting for the absorption of several tissue chromophores, provided excellent contrast between healthy tissue and tumor. Combining the results of spectral analysis with spatially resolved information provided a map of the tumor borders in greater detail than previously reported. The tumor dimensions determined with PA imaging were strongly correlated with those determined by histopathological examination for both melanomas and nevi.

Erratum to: Successful repair of a full upper eyelid defect following traumatic amputation by simply suturing it back in place, JPRAS Open volume 19, March 2019, pages 73-76.

[This corrects the article DOI: 10.1016/j.jpra.2018.12.002.].

Laser Speckle Contrast Imaging of a Rotational Full-Thickness Lower Eyelid Flap Shows Satisfactory Blood Perfusion.

Full-thickness eyelid flaps from the lower eyelid are frequently used to repair larger upper eyelid defects. Perfusion monitoring has recently been implemented in several reconstructive surgical procedures, however, perfusion monitoring of a rotational eyelid flap has not yet been described. The authors' employed laser speckle contrast imaging to monitor blood perfusion in a rotational flap from the lower eyelid, used to cover a large tumor defect in the upper eyelid. Perfusion in the flap decreased by only 50% during surgery and was almost completely restored 5 weeks later at flap division (91%). The excellent surgical outcome in the present case is deemed to be the result of satisfactory blood perfusion of the flap.

Novel Evidence Concerning Lacrimal Sac Movement Using Ultra-High-Frequency Ultrasound Examinations of Lacrimal Drainage Systems.

PURPOSE: Current hypothesis regarding the mechanism of active tear drainage is based on studies performed ex vivo or under nonphysiological conditions. Novel ultra-high-frequency ultrasound has the advantage of generating images with superior resolution, enabling measurements of low flow in small vessels, and the tracking of tissue motion in real time. The purpose of this study was to investigate the lacrimal drainage system and active drainage using this modality. METHODS: The upper lacrimal drainage system was investigated with 40-70 MHz ultrasound in 22 eyes in 13 patients. Irrigation confirmed a lacrimal obstruction in 10 eyes. Motion tracking was used to map movement of the lateral lacrimal sac wall and to measure flow when possible. RESULTS: The anatomy of the upper lacrimal drainage system was mapped in vivo, including the proximal canaliculi, which have not previously been imaged. The lacrimal sac lumen is slit shaped in its resting state but is distended when irrigated or if a nasolacrimal duct obstruction is present. Thus, the healthy lacrimal sac is not a cavity, and the medial retinaculum does not act against a stretched structure. Motion tracking visualized the "lacrimal pump," showing that the direction of motion of the lateral lacrimal sac wall is mainly in the sagittal plane during blinking. CONCLUSIONS: Ultra-high-frequency ultrasound allows detailed physiological monitoring of the upper lacrimal drainage system in vivo. Our findings suggest that current theories of active tear drainage need to be reappraised.

Erratum to "Blood perfusion in Hewes tarsoconjunctival flaps in pigs measured by laser speckle contrast imaging". JPRAS open, volume 18, December 2018, pages 98-103.

[This corrects the article DOI: 10.1016/j.jpra.2018.07.001.].

Unique spectral signature of human cutaneous squamous cell carcinoma by photoacoustic imaging.

Cutaneous squamous cell carcinoma (cSCC) is a common skin cancer with metastatic potential. To reduce reoperations due to nonradical excision, there is a need to develop a technique for identification of tumor margins preoperatively. Photoacoustic (PA) imaging is a novel imaging technology that combines the strengths of laser optics and ultrasound. Our aim was to determine the spectral signature of cSCC using PA imaging and to use this signature to visualize tumor architecture and borders. Two-dimensional PA images of 33 cSCCs and surrounding healthy skin were acquired ex vivo, using 59 excitation wavelengths from 680 to 970 nm. The spectral response of the cSCCs was compared to healthy tissue, and the difference was found to be greatest at wavelengths in the range 765 to 960 nm (P < .05). Three-dimensional PA images were constructed from spectra obtained in the y-z plane using a linear stepper motor moving along the x-plane. Spectral unmixing was then performed which provided a clear three-dimensional view of the distribution of tumor masses and their borders.

Perfusion in Upper Eyelid Flaps: Effects of Rotation and Stretching Measured With Laser Speckle Contrast Imaging in Patients.

PURPOSE: The aim of this study was to investigate how the blood perfusion in human upper eyelid skin flaps is affected by the length of the flap and the degree of stretching and rotation of the flap. METHODS: Twenty-nine upper eyelids were dissected as part of a blepharoplastic procedure in patients. The 1-cm wide proximal end of the flap remains attached, to mimic a random pattern skin flap (hereafter called a "skin flap"). Blood perfusion was measured with laser speckle contrast imaging before and after the flap was stretched with forces of 0.5, 1, and 2 N. The flap was then rotated 90°, and the same tensions were applied. RESULTS: Blood perfusion decreased gradually from the base to the tip of the flap. The flap was only well perfused in the proximal 1 cm (60% at 0.5 cm and 37% at 1.0 cm) and was minimally perfused beyond 2 cm (22% at 2.0 cm). Stretching the nonrotated flaps affected perfusion slightly (decreased to 43% at 0.5 cm). Simply rotating the flaps by 90° had no significant effect on the perfusion. The combination of rotation (90°) and stretching reduced the perfusion to 22% at 2 N, when measured 0.5 cm from the base. CONCLUSIONS: Blood perfusion in upper eyelid skin flaps decreases rapidly with distance from the base of the flap. Rotating and stretching the skin flap reduces blood perfusion even further, leading to minimal perfusion in this kind of flap at distances greater than 1.5 cm from the base.

Spectral Signatures in the Different Layers of the Human Eyelid by Photoacoustic Imaging.

BACKGROUND AND OBJECTIVES: The eyelids are susceptible to a number of skin cancers, which are challenging to excise radically without sacrificing excessive healthy tissue. Photoacoustic (PA) imaging is an emerging non-invasive biomedical imaging modality that could potentially be used for intraoperative micrographic control of the surgical margins of eyelid tumors. In this study, non-cancerous human eyelid tissue was characterized using PA as a first step in the development of this technique. STUDY DESIGN/MATERIALS AND METHODS: Twelve full-thickness samples from nine patients were analyzed ex vivo using PA imaging. Two-dimensional PA images were acquired using 59 wavelengths in the range of 680-970 nm to obtain the spectral signatures of the skin, orbicularis oculi muscle, and the tarsal plate. Three-dimensional images were obtained by scanning the tissues using a linear stepping motor. Spectral unmixing was performed to visualize the chromophore distribution. RESULTS: The resulting PA spectra could be used to differentiate between the orbicularis oculi muscle and the other two structures (P < 0.05). The signals from the skin and the tarsal plate were more similar in appearance, probably due to similarities in their molecular composition. Spectral unmixing provided a clear visualization of the overall architecture of the eyelids. CONCLUSIONS: PA imaging can be used to differentiate between the orbicularis oculi muscle and the eyelid skin and tarsal plate. The main structures of human eyelids could be visualized in three dimensions using PA imaging. This technique could potentially be used to examine eyelid tumors intraoperatively in the future. However, further studies on tumors in vivo are needed before considering such clinical use. Lasers Surg Med. © 2019 Wiley Periodicals, Inc.

The Effect of Canthotomy on Blood Perfusion During the Repair of Lower Eyelid Defects.

PURPOSE: Canthotomy is frequently used to mobilize extra tissue when repairing larger lower eyelid defects. The aim of this study was to explore the effect of canthotomy on blood perfusion and oxygen tension. METHODS: Eight pigs underwent a wedge resection of the lower eyelid and canthotomy (with cantholysis involving the lateral palpebral artery). The wedge resection was performed 8, 6, and 4 mm from the canthotomy. Perfusion and oxygen tension were monitored in the eyelid between the wedge resection and canthotomy using laser Doppler velocimetry and a Clark electrode. Verapamil was administered, and measurements were also performed 12 hours after surgery, to investigate the possible effects of vasospasm RESULTS:: The wedge resection alone did not affect perfusion. Canthotomy led to a reduction in perfusion; being 60% when the length of remaining eyelid was 8 mm, 32% when it was 6 mm, and 24% when it was 4 mm. Similar results were observed for oxygen tension. Vasospasm did not affect the results. CONCLUSIONS: Canthotomy in combination with a wedge resection of the lower eyelid affects blood perfusion. A smaller length of remaining eyelid tissue will have less perfusion. This may not have any implications in cases of direct closure, but may play a role when the eyelid is to provide oxygen and nutrients to avascular grafts.

Blood Perfusion in Rotational Full-Thickness Lower Eyelid Flaps Measured by Laser Speckle Contrast Imaging.

PURPOSE: Large upper eyelid defects can be repaired by rotational full-thickness lower eyelid flaps. The aim was to measure the blood perfusion in such flaps, and how it is affected by the length of the flaps, and the degree of rotation and stretching. METHODS: Nine patients underwent the Quickert procedure for entropion repair in which a full-thickness eyelid flap of approximate width 0.5 cm and length 2 cm was dissected in the lower eyelid. This generates a full-thickness eyelid flap similar to that used to repair large upper eyelid defects. Perfusion was measured using laser speckle contrast imaging, before and after the flap was rotated 90° and 120°, and stretched using forces of 0.5, 1, and 2 N. RESULTS: Blood perfusion decreased gradually from the base to the tip of the flap; being 75% of the reference value 0.5 cm from the base, 63% at 1.0 cm, 55% at 1.5, 23% at 1.75 cm, and 4% at 2.0 cm. Rotating the flaps by 90° or 120° had little effect on the perfusion. Stretching reduced the perfusion from 63% to 32% at 2 N, when measured at 1 cm. The combination of stretching and rotation did not lead to any further decrease. CONCLUSIONS: Blood perfusion in lower eyelid rotational flaps seems to be more sensitive to stretching than to rotation. Provided the flap is no longer than 1.5 cm, the perfusion will be greater than 20%, even when rotated, which should be sufficient for adequate survival and healing.

Revascularization of Free Skin Grafts Overlying Modified Hughes Tarsoconjunctival Flaps Monitored Using Laser-Based Techniques.

PURPOSE: It has recently been shown that the flap pedicle does not supply blood to a tarsoconjunctival graft in the modified Hughes procedure in patients. This raises questions concerning the rate of revascularization of the free skin graft commonly used to reconstruct the anterior lamella. The aim of this study was, thus, to monitor the course of revascularization in free skin grafts overlying modified Hughes tarsoconjunctival flaps, using laser-based techniques. METHODS: Free skin grafts from the upper eyelid or upper arm in 9 patients were used to cover a tarsoconjunctival flap according to the modified Hughes procedure. Blood perfusion was monitored using laser speckle contrast imaging, and vascular reactivity was studied with laser Doppler velocimetry after heating the tissue to 44°C. Measurements were made at the time of surgery (baseline) and at 1, 3, 8, and 16 weeks postoperatively. RESULTS: The gradual increase in perfusion of the free skin grafts during the healing process indicates revascularization. A slight increase in perfusion was seen already after 1 week. Perfusion reached 50% of the baseline after 3 weeks, and complete restoration of perfusion was seen after 8 weeks. The vascular function monitored with heat-induced hyperemia increased in a similar fashion. CONCLUSIONS: Full-thickness skin grafts revascularize within 3 to 8 weeks, despite overlying a tarsoconjunctival flap, which has recently been reported to be avascular. This provides further evidence that it should be possible to repair large eyelid defects using free full-thickness eyelid grafts.

Successful repair of a full upper eyelid defect following traumatic amputation by simply suturing it back in place.

There is a general belief that a full-thickness eyelid defect is best repaired using a vascularized flap in combination with a free graft, and that a free full-thickness eyelid graft would not survive due to poor blood perfusion. However, we describe a case in which an upper eyelid was traumatically amputated. The eyelid was sutured in place and healed well in situ. The long-term outcome was good regarding motility and function. This raises the question of whether a blood-supplying pedicle is necessary for the survival of the graft when repairing large eyelid defects.

Maximal haemostatic effect is attained in porcine skin within 7 min of the administration of a local anaesthetic together with epinephrine, refuting the need for a 30 min waiting time.

OBJECTIVE: Based on clinical experience gained over many years, the maximal haemostatic effect following administration of local anaesthetics containing epinephrine is generally believed to occur within 10 min. Surprisingly, it was found in a recent study, in which bleeding was quantified intraoperatively, that maximal haemostasis did not occur until 30 min. If this is indeed the case, then it would be necessary to extend the preoperative waiting time to minimize perioperative bleeding.We have carried out a carefully controlled study on the time delay between administration of a local anaesthetic containing epinephrine and maximal haemostasis in a surgical setting. METHODS: Lidocaine 20 mg/ml (2%) or lidocaine + epinephrine 12.5 µg/ml (1:80,000) was injected into the skin of eight pig flanks. Bleeding was induced after 3, 5, 7, 15 and 30 min by making a 10 mm incision at each injection site. Blood was collected for 1 min and weighed. RESULTS: A gradual reduction in bleeding was observed, with maximal reduction after only 7 min (54%, p < 0.05, 95% CI: 44-64%). No further significant reduction in bleeding was observed (62% at 15 and 66% at 30 min, p = n.s. compared to 7 min). CONCLUSIONS: Maximal haemostatic effect in the current setting was observed within 7 min of injection of lidocaine with epinephrine. This is in good agreement with previous empirical findings, and we see no reason to prolong the preoperative waiting time.

Hypoperfusion following the injection of epinephrine in human forearm skin can be measured by RGB analysis but not with laser speckle contrast imaging.

BACKGROUND: The time taken for epinephrine to achieve its optimal effect during local anesthesia has recently become the subject of debate. The time from injection to commencement of surgery is traditionally quoted to be 7 to 10 min, while recent reports claim that it may take 30 min to achieve maximum hypoperfusion, which would prolong the time required for surgical procedures. The discrepancy may be related to difficulties associated with the techniques used to measure blood perfusion. The aim of this study was to test two methods of determining the time to maximum hypoperfusion. METHODS: Laser speckle contrast imaging (LSCI) and red, green, blue (RGB) analysis of images obtained with a commercial digital camera, were used to monitor the effect of infiltration with commonly used local anesthetic preparations: lidocaine (20 mg/ml) + epinephrine (12.5 µg/ml), lidocaine (10 mg/ml) + epinephrine (5 µg/ml), and lidocaine (20 mg/ml) alone, in healthy subjects. RESULTS: LSCI showed a paradoxical increase in signal after the injection of local anesthetics containing epinephrine, probably due to a change in the laser penetration depth resulting from blanching of the skin. However, RGB analysis of digital photographs gave more reliable results, showing skin blanching that corresponded to the expected effect of epinephrine in local anesthetics. The time to maximum effect was found to be 7 (range 5-10) minutes for 12.5 µg/ml epinephrine, and 9 (range 7-13) minutes for 5 µg/ml epinephrine in lidocaine. CONCLUSIONS: RGB analysis of digital images proved to be a valid technique for monitoring the effect of local anesthetics with epinephrine in human skin. The technique requires only a commercial digital camera and constitutes a cheap, simple method. The optimal delay between epinephrine injection and incision, to minimize bleeding, was found to be 7 to 9 min, which is in good agreement with common surgical practice.

Perfusion Monitoring Shows Minimal Blood Flow From the Flap Pedicle to the Tarsoconjunctival Flap.

BACKGROUND: A previous study in pigs has shown that the pedicle of the tarsoconjunctival flap does not appear to have adequate blood perfusion. The aim of this study was to monitor perfusion in tarsoconjunctival flaps in patients with large lower eyelid defects resulting from tumor surgery. METHODS: The modified Hughes procedure was performed in 13 patients. Blood perfusion was monitored using laser Doppler velocimetry and laser speckle-contrast imaging. RESULTS: Blood flow decreased gradually from the pedicle base to the end of the flap and was 19% at the flap base, 11% in the middle of the flap, and 4% in the distal end of the flap. The flaps survived, and there was no tissue necrosis. CONCLUSIONS: Tarsoconjunctival tissue survival does not seem to be dependent on a conjunctival flap. Free tarsoconjunctival grafts or composite grafts might be considered as viable alternatives in reconstruction of major eyelid defects.