Project title:

“Development and clinical validation of a novel cost effective multi-modal methodology for early diagnostics of skin cancers.”

Project contract number:

1.1.1.2/VIAA/1/16/052

Project partners:

University of Oulu, Faculty of Electrical Engineering, Laboratory of Optoelectronics and measurement techniques.

Bulgarian Academy of Sciences, Academician Emil Djakov Institute of Electronics, Laboratory of Biophotonics.

Spectral Molecular Imaging, Inc. Beverly Hills, California.

Project implementation deadline:

01.01.2018. - 31.12.2020.

Total project funding:

133 805,88 EUR

ERAF funding:  113 734,99 EUR 

State budget financing: 13 380,58 EUR 

UL funding: 6 690,31 EUR

Research manager:

Ilze Ļihačova (Biofotonikas laboratorija)

Aim of project:

To develop and clinically test a new, inexpensive multimodal methodology for early diagnosis of skin cancers (melanoma, basal cell and squamous cell carcinoma), postoperative scarring and early detection of tumor recurrence.

Acknowledgement for publications:

This work has been supported by European Regional Development Fund project “Development and clinical validation of a novel cost effective multi-modal methodology for early diagnostics of skin cancers” (No. 1.1.1.2/16/I/001, agreement No. 1.1.1.2/VIAA/1/16/052).

Projekta rezultāti: 

2 scientific articles in international peer-reviewed journals with calculated citation index, 4 conference abstracts, 1 popular science article, 3 mobility trips, 3 scientific networking.

Publications:

  1. A. Lihachev, I. Lihacova, E. V. Plorina, M. Lange, A. Derjabo, and J. Spigulis, "Differentiation of seborrheic keratosis from basal cell carcinoma, nevi and melanoma by RGB autofluorescence imaging," Biomed Opt Express 9(4), 1852-1858 (2018). DOI 10.1364/BOE.9.001852
  2. I. Lihacova, K. Bolochko, E. V. Plorina, M. Lange, A. Lihachev, D. Bliznuks, and A. Derjabo, "A method for skin malformation classification by combining multispectral and skin autofluorescence imaging," SPIE Proc 10685, 1068535 (2018). DOI 10.1117/12.2306203.
  3. K. Bolochko, D. Bliznuks, I. Lihacova, and A. Lihachev, "Quality enhancement of multispectral images for skin cancer optical diagnostics," SPIE Proc 10679, 1067903 (2018). DOI 10.1117/12.2306609.
  4. A. Sdobnov, A. Bykov, A. Popov, I. Lihacova, A. Lihachev, J. Spigulis, and I. Meglinski, “Combined multi-wavelength laser speckle contrast imaging and diffuse reflectance imaging for skin perfusion assessment,” SPIE Proc 11075, Novel Biophotonics Techniques and Applications V, 110751F (2019); https://doi.org/10.1117/12.2526921.
  5. Gunita Nagle “Kā izķert melanomu,” IR, (Jun 12, 2019), https://ir.lv/2019/06/12/ka-izkert-melanomu/?fbclid=IwAR2vntxGelERu8BtzAaXFsHZCNAsf2egWG0kZPCHd464GeHRx8nh3iBCg7I
  6. Ilze Ļihačova, Aleksejs Ļihačovs, Dmitrijs Bļizņuks, "Portable device for early noninvasive diagnosis of skin cancer," Latvian Academy of Sciences Yearbook 2020, 56 pp., http://www.lza.lv/images/stories/YearBook_2020.pd

1st quarter (01.01.2018 - 31.03.2018)

Literature study on the applications of circularly polarized light for the analysis of skin formations. Investigation of various experimental schemes using circularly polarized lighting and planning of an experiment on the reflection of circularly polarized light from the skin surface. Planning of the existing prototype for obtaining multispectral images, transformation for the analysis of skin surfaces illuminated with circularly polarized light. Addition of the developed and published skin autofluorescence method to the existing p 'map analysis method in order to obtain a more sensitive multimodal methodology for skin cancer diagnosis. Analysis of multispectral images and autofluorescence images has been initiated for various groups of skin pathologies not previously studied by the p 'diagnostic method - seborrheic keratoses, hyperkeratoses, unpigmented basal cell carcinomas and hemangiomas - as well as melanocytic nevus and melanomas. A method for the analysis of the maximum values ​​of p 'maps was introduced and applied to the data of the existing skin formation groups. Mean values ​​of autofluorescence intensities for different groups of skin formations were calculated. The two methods - the analysis of the maximum values ​​of the p 'maps and the autofluorescence imaging method - were combined. Thus, a new diagnostic parameter with 100% sensitivity and 100% specificity to distinguish melanoma from other groups of skin formations was created. With this method it is not possible to distinguish basal cell carcinoma from groups of benign skin formations. One article was published in the journal Biomedical Optics Express, 2 articles were submitted for SPIE Proceedings conference materials.

2nd quarter (01.04.2018 - 30.06.2018)

  1. A report presentation of the first quarter of the Postdoc project was prepared, and on April 19 it was presented at the IAPS UL seminar.
  2. Participation in the SPIE Photonics 2018 conference (April 23 - 26, Strasbourg, France):
    • Preparation and presentation of the poster report "A method for skin malformation classification by combining multispectral and skin autofluorescence imaging"; Thesis available: www.spie.org/EPE/conferencedetails/biophotonics-photonic-solutions
    • Co-author of the oral report “Quality enhancement of multispectral images for skin cancer optical diagnostics.” Thesis available: www.spie.org/EPE/conferencedetails/optics-photonics-digital-technologies-multimedia-applications
    Two articles have been published in the journal SPIE Proceedings.
  3. Mobility cooperation agreements prepared, signed and forwarded to the University of Oulu partner.
  4. An in-depth study has been carried out on optical aids that could be used to obtain circularly polarized radiation for further experiments. As a result, it was decided to use a polymer film (it is easier to adapt to the experiment) to obtain circular EM radiation in the range of 400 - 700 nm and to test whether it will be possible to obtain any detectable tissue structure properties in this region. Data were selected for further calculations from newly acquired measurements in the LOC clinic using linearly polarized radiation.

3rd quarter (01.07.2018 - 30.09.2018)

  • A device has been developed for the study of the reflection of circularly polarized light from the skin surface. The created device consists of a multispectral camera Nuance EX, which consists of a linear polarizer, a light source (halogen lamps + LEDs) and a film for obtaining circular electromagnetic radiation, which is placed in the path of the light source and converts non-polarized light into circularly polarized. To test how different skin structures reflect polarized light at different angles, the film angle was changed in 45 ° increments and images were taken in the spectral range from 450 to 950 nm (in 10 nm increments). The result was multispectral data for whole skin, benign nevus and seborrheic keratosis measured at 8 different angles (0 °, 45 °, 90 °, 135 °, 180 °, 225 °, 270 ° and 315 °).
  • Thesis was submitted: Janis Spigulis, Uldis Rubins, Edgars Kviesis-Kipge, Andris Grabovskis, Aleksejs Lihacovs, Ilze Lihacova, Dmitrijs Bliznuks, Ilona Kuzmina, Ilze Oshina, Marta Lange, “Advanced imaging technologies for distant assessment of in-vivo skin,” in which I co-authored the Northern Optics & Photonics Conference in Lund, Sweden, September 12 - 14, 2018.
  • July 30 to September 23 - vacation.
  • Published thesis: Janis Spigulis, Uldis Rubins, Edgars Kviesis-Kipge, Andris Grabovskis, Aleksejs Lihacovs, Ilze Lihacova, Dmitrijs Bliznuks, Ilona Kuzmina, Ilze Oshina, Marta Lange, “Advanced imaging technologies for distant assessment of in-vivo skin” Northern Optics & Photonics  2018 conference proceedings. The thesis is available here: photonicsweden.org/wp-content/uploads/NorthernOptics2018_proceedings.pdf
  • An informative presentation and poster about the project and its current and planned activities have been prepared. Participation in the Scientists' Night on September 28, the program is available at http://www.asi.lu.lv/New/programma2018.pdf.

4th quarter (01.10.2018 - 31.12.2018)

  • Mobility to the University of Oulu in Finland (8 - 19 October). The University of Oulu, Optoelectronics and Measurement Techniques Laboratory, was visited under the guidance of Igor Meglinski. Networking, presentation of the goals, tasks and methodologies of the project “Development of a new and cost-effective multimodal methodology and clinical approbation for early diagnosis of skin cancer” was carried out. A tour of the laboratory was visited, working tables of various experiments were performed, including tables for circular and linear polarization measurements. It was found that circular polarization is not so easily "disrupted" compared to linear polarization, therefore, by correctly detecting the reflected radiation, it is possible to obtain information from the deeper layers of the skin, information about collagen. Collagen imaging could greatly improve the existing methodology, as melanoma collagen structures are known to be more heterogeneous than in healthy skin and other skin formations. It was found out what literature to study to better understand circular polarization. Mr Meglinski was interested in the diagnostic criterion for melanoma that I developed and together we discussed its physical explanation. As a result, we decided to study this parameter more closely and, in collaboration with Anton Sdobnov (University of Oulu), to perform an additional study using contrast analysis of laser beams.
  • A concept for the next skin formation diagnostic prototype was developed. Including circular polarization. As a result of networking, it was found that circularly polarized radiation, when reflected from the skin surface, changes its direction of rotation in the opposite direction, but when reflected from collagen in the skin, it does not change its direction of rotation, so the first detector information on the structure of collagen in the skin.
  • Work has been done to differentiate basal cell carcinoma from benign skin formations. Correction was first made in the green and red channels based on white reference measurements taken immediately prior to skin formation measurements. It was hypothesized that basal cell carcinomas may have a different slope of the line drawn between the green channel intensities and the red channel intensities.
  • A mobility cooperation agreement with the Bulgarian Academy of Sciences was prepared, signed and forwarded.
  • A script was developed for the correction of clinical measurement data using linear polarization (for the green and red channels) and for the calculation of the slope coefficient (between the G and R channel intensities). Started linear polarized data processing - slope coefficient calculations. The first results did not show a significant difference between basal cell carcinomas, nevus and hyperkeratosis. It is planned to supplement the data set and to include in the calculations only basal cell carcinomas, which are also confirmed histologically or cytologically.
  • From December 10 to 24, incapacity for work.
  • Report (Milestone M1.1) on the application of circularly polarized radiation to low-cost skin imaging techniques for future medical applications and Deliverable D1.1. or a summary of the method for determining the optical properties of the skin.

5th quarter (01.01.2019 - 31.03.2019)

  • 02.01.2019. - 13.01.2019 incapacity for work.
  • A presentation on the work done in 2018 was prepared (presented on January 8 at the meeting of the University of Latvia, presented by A. Lihačov due to incapacity for work).
  • Submitted paper: A. Sdobnov, A. Bykov, A. Popov, I. Lihacova, A. Lihachev, J. Spigulis, and I. Meglinski, “Combined multi-wavelength laser speckle contrast imaging and diffuse reflectance imaging for skin perfusion assessment” European Conferences on Biomedical Optics (ECBO) to be held on 23-27 June in Munich, Germany.
  • On January 24, participation in the IAPS UL seminar with a presentation for the position of the leading researcher at the IAPS UL, where the current activities and results of the project are presented.
  • On February 8, a letter of thanks was received from the President of the Latvian Academy of Sciences for the 2018 achievement in applied science for the submitted achievement “Portable device for early contactless diagnosis of skin cancer”, which includes the methodology developed in the postdoctoral project.
  • On February 20, a presentation on this achievement was given entitled "Portable device for early contactless diagnosis of skin cancer."
  • Prepared and submitted conference paper: Marta Lange, Emilija Vija Plorina, Ilze Lihacova, Janis Spigulis, “Skin cancer screening - better safe than sorry,” SHS Web of Conferences, 7th International Interdisciplinary Scientific Conference SOCIETY. HEALTH. WELFARE.
  • On February 14 - 22, networking with M.Sc. Martin Kolar from Brno University of Technology, Czech Republic. The problems of the methodology developed in the postdoctoral project were explained in detail and possible further cooperation opportunities were found. Coordination of experiments planned for the mobility trip, identification of inventory, discussion of samples with a contact person (E. Borisova) from the Institute of Electronics, Bulgarian Academy of Sciences.
  • March 24 - 29 mobility trip to Emil Djakov Institute of Electronics, Bulgarian Academy of Sciences, Biophotonics Laboratory. During the visit, measurements were performed using an 808 nm, 100 mW laser, NIRQuest 2.1. (from 900 to 2100 nm) and USB4000 (200 to 1100 nm) spectrometers. Measurements were performed on a nevus sample ex-vivo. No IR autofluorescence was observed. In addition, measurements were made using a 785 nm, 100 mW laser and an OceanOptics USB mini spectrometer. Infrared autofluorescence spectra were obtained for various skin pathologies - unpigmented and pigmented basal cell carcinomas, pigmented nevus, dysplastic nevus and melanoma. A postdoctoral project was presented at a scientific seminar, as well as networking with Bulgarian scientists.

6th quarter (01.04.2019 - 30.06.2019)

  • From April 3 to 5, the World Congress on Advanced Treatments and Technologies in Skin Cancer was held in Vienna, Austria, to learn about the latest trends in skin cancer diagnosis and treatment.
  • Infrared autofluorescence data (excitation with a 785 nm 100 nW laser) were analyzed in April. Data were obtained at the Institute of Electronics in Bulgaria from various ex-vivo skin samples (basal cell carcinoma, squamous cell carcinoma, melanoma, dysplastic nevus, nevus - various skin layers) stored in formaldehyde. After normalization, all spectra showed the same shape, so it was chosen to analyze the intensities of the samples. Pigmented skin formations were found to have a lower autofluorescence intensity than non-pigmented ones, which is contrary to the literature. It was also found that the different skin layers of the nevus - epidermis, dermis and hypodermis have very similar intensities, which indicates that there is no direct correlation with the intensity with melanin concentration. In contrast, in vivo measurements in the skin showed that the surface of the finger with a higher concentration of melanin has a higher intensity of autofluorescence than the underside of the fingertip, which is in agreement with the results available in the literature, but does not agree with the ex-vivo sample data. On April 25, the results obtained in Bulgaria were presented at the IAPS UL seminar.
  • In May, a communication was conducted with Bulgarian and Russian scientists to interpret the obtained autofluorescence results. It was concluded that these experiments would require a filter with a higher optical density OD > 10. As the laser line was not completely blocked by the selected filter, the data are difficult to interpret.
  • The following publication was prepared and submitted: A. Sdobnov, A. Bykov, A. Popov, I. Lihacova, A. Lihachev, J. Spigulis, and I. Meglinski, “Combined multi-wavelength laser speckle contrast imaging and diffuse reflectance imaging for skin perfusion assessment. "
  • Vacation from May 7 to 16.
  • On May 17, a postdoctoral project was presented at the event "Knowledge Agora" organized by the University of Latvia.
  • From May 20. - 2 June leave.
  • On May 29, the "REGIOSTARS" competition award was received, where the project "Portable device for early contactless diagnosis of skin cancer", based on multi-modal methods developed in the postdoctoral project, won first place (https://regiostars.lv/portfolio-item/1-vieta-portativa-ierice-adas-veza-agrinai-bezkontakta-diagnostikai-projekta-pieteicejs-latvijas-universitate/?fbclid=IwAR24pWBzbyTFo48HtEgMNmHVefoptjgijgs9RSiocc-wi1b7rHbflBV8BGI ).
  • An interview was given to IR magazine. A popular scientific article has been published: Gunita Nagle “How to catch melanoma,” which mentions the development, topicality and results of the method developed in the postdoctoral project.
  • Additional polarization experiments were performed on a group of 10 nevus. Multispectral images were obtained using 4 different polarization methods, where linear polarizers were placed 1) parallel, 2) perpendicular to the light source and receiver, and where 3) equally directed and 4) counterclockwise circular polarizers placed opposite the light source and the receiver were used.
  • From 18 to 19 June, a networking event for young researchers from the Baltic region ("Engaging in research policy discussions. An event for early career researchers from the Baltic region") was attended at the Tallinn Academy of Sciences. At the event, I gave a speech outlining the main shortcomings related to the implementation of projects. Networking with young scientists not only from the Baltic region, but also from other parts of Europe. Information about the event is available here: https://www.facebook.com/pg/teadusteakadeemia/photos/?tab=album&album_id=2380985112146714&__tn__=-UCH-R.
  • From 23 to 27 June, participation in the European Conference on Biomedical Optics with a report: A. Sdobnov, A. Bykov, A. Popov, I. Lihacova, A. Lihachev, J. Spigulis, and I. Meglinski, “Combined multi- wavelength laser speckle contrast imaging and diffuse reflectance imaging for skin perfusion assessment. ” The conference thesis is available in the SPIE application and will also be available here: https://spie.org/conferences-and-exhibitions/european-conferences-on-biomedical-optics?utm_id=rebo19gb. During the conference, presentations and events were attended, as well as networking with young scientists in the field of biomedical optics.
  • Milestone 2.1. Preparation of "Recommendations for NIR AF display using LEDs and RGB / CMOS sensors".

7th quarter (01.07.2019 - 30.09.2019)

  • Due to the ambiguity of the results obtained at the Bulgarian Academy of Sciences, Institute of Electronics, a NIR autofluorescence imaging system was developed to determine whether it is possible to obtain NIR-induced skin AF. Experiments were performed using an 808 nm laser diode light source, an RGB / CMOS camera and an 825 nm, OD ≥ 4 long pass filter. As the OD of the existing filter at 808 nm is lower (OD808 = 4) than in the measurements made in Bulgaria with a 785 nm laser (at 785 nm for the specific filter OD785 = 5), it was not possible to filter the laser excitation radiation sufficiently and the skin autofluorescence signal could not be obtained.
  • From 01.08.2019. - 30.08.2019 annual vacation.
  • An experimental stand for imaging autofluorescence of skin induced by near infrared radiation was designed. A Raman long pass 785 nm filter with OD ≥ 6, CMOS camera will be used in the experimental bench. Participation with an informative poster about the activities of the postdoctoral project on Scientists' Night 27.09.2019.

8th quarter (01.10.2019 - 31.12.2019)

  • An experiment was performed in October to obtain near-infrared-induced autofluorescence images of the skin. An 808 nm laser diode, an Edmund Optics 825 nm longpass filter and a CMOS camera were used in the experiment. No autofluorescence was observed, probably due to the filter not completely blocking (OD ≥ 4) the laser diode radiation. A 760 nm LED, a spectrometer and a changed filter to a 785 nm Raman longpass edge filter with a higher OD (OD ≥ 6) were used to obtain skin autofluorescence. As the 760 nm diode has a very wide emission band (diode emission was also observed after 785 nm), LED radiation could obscure the weak autofluorescence. To better understand near-infrared-induced autofluorescence of the skin, an expert Ivan Bratchenko was invited to the laboratory, who has succeeded in obtaining Raman spectra of skin autofluorescence and claims that the skin has autofluorescence induced by near-infrared radiation. An interview was given to the Latvian Television program “Cognitive Impulse” (Part 2 of the program of 21.11.2019: https://ltv.lsm.lv/lv/raksts/21.11.2019-izzinas-impulss.id173231/LU.
  • In November, an experiment using 660 nm LEDs was performed to avoid LED radiation beyond the 785 nm Raman long pass filter. A 785 nm Raman longpass edge filter and spectrometer were used in the experiment, but still no autofluorescence was observed. From November 11 to 15, doctoral student Deyan Ivanov from a project partner institution in Bulgaria (Bulgarian Academy of Sciences, Institute of Electronics, Sofia, Bulgaria) was admitted to the laboratory to obtain near infrared-induced autofluorescence from skin samples in formalin. In collaboration with researcher Mindaugas Tamošiūnas from Vytautas Magnus University (Vilnius, Lithuania), pigmented and unpigmented skin formations were measured using a Raman spectroscopy system (with 785 nm laser excitation). During Deyan Ivanov's visit, experiments as well as networking activities were carried out. From November 18 to 23, scientist Ivan Bratchenko, an expert in the measurement of near-infrared-induced autofluorescence in the skin using the Raman spectroscopy system (with 785 nm laser excitation), was admitted to the laboratory. In collaboration with Lithuanian scientist Mindaugas Tamošiūnas, melanoma and benign cells were prepared, the spectra of which were recorded using the Raman spectroscopy system. In addition, Raman spectra of skin and pigmented nevus were recorded. Scientific networking activities were carried out.
  • In December, a popular scientific article by I. Lihacova, A. Lihachev, D. Bliznuks “Portable device for early noninvasive diagnosis of skin cancer” was prepared and submitted to the Yearbook of the Latvian Academy of Sciences 2020. Data on melanoma and non-melanoma cells obtained in December were processed. In the first approximation, the most significant differences between the near-infrared-induced autofluorescence spectra of melanoma and non-melanoma cells that could be used for imaging were observed in the spectral range of 940 – 960 nm.

9th quarter (01.01.2020 - 31.03.2020)

  • In January, Raman spectra obtained from melanoma and non-melanoma cells were analyzed. The results suggest that there is no clear correlation between melanoma and non-melanoma 785 nm induced autofluorescence intensities. The shape of the autofluorescence spectrum of melanoma cells corresponds to I. Bratchenko et al. 2017 published results, which allow to distinguish melanoma cells from non-melanoma cells using near-infrared-induced form of autofluorescence spectrum in the range from 800 to 900 nm. In order to distinguish melanoma cells from non-melanoma cells, it is necessary to obtain the shape of the near-infrared-induced autofluorescence spectrum, but this method is not suitable for a fast, simplified imaging technique. The obtained results were presented at the UL conference in 2020. February 14.
  • In February, applications were prepared and submitted to two conferences: I.Lihacova, A. Lihachev, M.Lange, E.V. Plorina, E. Cibulska, D. Bliznuks, A. Derjabo, N. Kiss, “Non-invasive multimodal imaging system for skin cancer detection,” “The 2nd World Congress on Advanced Treatments in Skin Cancer (Skin-Cancer2020)” in Berlin , Germany, April 2-3. Due to Covid-19, the Congress was canceled indefinitely, but the abstract was adopted, it will be presented as soon as possible. I.Lihacova, A. Lihachev, M.Lange, E.V. Plorina, E. Cibulska, D. Bliznuks, A. Derjabo, N. Kiss, “Optical multimodal non-invasive diagnostics of skin cancer,” “International Conference on Biomadical Photonics 2020,” Montpellier, France, 16-18. in April. The abstract was adopted, but due to Covid-19, the conference was canceled and carried over to the following year. Participation in the 78th LU conference in 2020. On February 14, the 2nd year report of the project was presented with a poster presentation on the results of the 2nd year of the project, as well as the evaluation of the Scientific Council.
  • In March, a cooperation agreement was prepared between the IAPS University of Latvia and the Venereology, Dermatology and Oncology Center in Semmelveis, Hungary, where measurements with a multimodal imaging prototype will be performed as soon as possible. Work has begun on collecting and organizing the accumulated data, selecting and preparing useful data for automatic reading and processing, as well as work on the development of image processing software for the segmentation of whole skin / skin formation.

10th quarter (01.04.2020 - 30.06.2020)

  • Leave from 1.04.2020. - 30.04.2020.
  • For automatic calculation of multispectral p 'parameter and primary diagnosis of melanoma, skin, marker and formation segmentation algorithm is developed using artificial neural networks. So far, a marker recognition algorithm has been developed and work is underway to recognize whole skin.
  • In parallel, a database read algorithm has been developed that reads data obtained at different times during prototyping and imaging and differs in the name creation and measurement structure. Work has begun on the creation of p 'and autofluorescence maps and value calculations, automatic result output.
  • Two theses were submitted for the conference Biophotonics Riga 2020, which will take place on August 24-25, Riga, Latvia: 1) I. Lihacova, A. Lihachev, M. Lange, EV Plorina, E. Cibulska, D. Bliznuks, A. Derjabo N. Kiss, “Optical multimodal non-invasive diagnostics of skin cancer.” 2) A. Lihachev, I. Lihacova, M. Lange, E. V. Plorina, E. Cibulska, D. Bliznuks, A. Derjabo, N. Kiss, “Imaging of LED excited autofluorescence for skin lesions assessment.”
  • Published in the popular scientific article "Portable device for early diagnosis of skin cancer", Latvian Academy of Sciences Yearbook 2020, 56 p. Available: http://www.lza.lv/images/stories/YearBook_2020.pdf
  • Multispectral data collected at the Latvian Oncology Center in Riga are used for clinical validation of the methodology. There is also a collaboration with the Faculty of Dermatology, Venereology and Dermatooncology, University of Semmelweiss, Budapest, Hungary, where clinical measurements with a prototype developed in synergy with the ERDF project “Portable Device for Early Contactless Diagnosis of Skin Cancer” and multispectral data storage are also performed. At the moment, data collection at the Latvian Oncology Center is not taking place due to the consequences of covid-19, but measurements have resumed in Budapest.