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Correction: 10.1039/d1na90120f Studies of extracellular vesicles (EVs), their trafficking and characterization often employ fluorescent labelling. Unfortunately, little attention has been paid thus far to a thorough evaluation of the purification of EVs after labelling, although the presence of an unbound dye may severely compromise the results or even lead to wrong conclusions on EV functionality. Here, we systematically studied five dyes for passive EV labelling and meticulously compared five typical purification methods: ultracentrifugation (UC), ultracentrifugation with discontinuous density gradient (UCG), ultrafiltration (UF), size exclusion chromatography (SEC), and anion exchange chromatography (AEC). A general methodology for evaluation of EV purification efficiency after the labelling was developed and tested to select the purification methods for the chosen dyes. Firstly, we found that some methods initially lead to high EV losses even in the absence of the dye. Secondly, the suitable purification method needs to be found for each particular dye and depends on the physical and chemical properties of the dye. Thirdly, we demonstrated that the developed parameter E-rp (relative purification efficiency) is a useful tool for the pre-screening of the suitable dye-purification method combinations. Additionally, it was also shown that the labelled EVs properly purified from the unbound dye may show significantly reduced contrast and visibility in the target application, e.g. in the live cell fluorescence lifetime imaging.

Hyperspectral imaging (HSI) applications for biomedical imaging and dermatological applications have been recently under research interest. Medical HSI applications are non-invasive methods with high spatial and spectral resolution. HS imaging can be used to delineate malignant tumours, detect invasions, and classify lesion types. Typical challenges of these applications relate to complex skin surfaces, leaving some skin areas unreachable. In this study, we introduce a novel spectral imaging concept and conduct a clinical pre-test, the findings of which can be used to develop the concept towards a clinical application. The SICSURFIS spectral imager concept combines a piezo-actuated Fabry–Pérot interferometer (FPI) based hyperspectral imager, a specially designed LED module and several sizes of stray light protection cones for reaching and adapting to the complex skin surfaces. The imager is designed for the needs of photometric stereo imaging for providing the skin surface models (3D) for each captured wavelength. The captured HS images contained 33 selected wavelengths (ranging from 477 nm to 891 nm), which were captured simultaneously with accordingly selected LEDs and three specific angles of light. The pre-test results show that the data collected with the new SICSURFIS imager enable the use of the spectral and spatial domains with surface model information. The imager can reach complex skin surfaces. Healthy skin, basal cell carcinomas and intradermal nevi lesions were classified and delineated pixel-wise with promising results, but further studies are needed. The results were obtained with a convolutional neural network. peerReviewed

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A spectral imaging device (500) comprises: - an optical modifier system (SYS1) to form axial light beams (LB2) from received light beams (LB1), the axial light beams (LB2) being parallel with an optical axis (AX1) of the imaging device (500), - a Fabry-Perot interferometer (FPI) to provide filtered axial light beams (LB3) by filtering light of the axial light beams (LB2), - an image sensor (SEN1), and - an array (ARR1) of lenses (LNS0,0, LNS0,1) to form a plurality of sub-images (S0,0, S0,1) on the image sensor (SEN1) by focusing light of the filtered light beams (LB3).Patent family as of 30.12.2022FI129998 B20221215FI2020000540520200422 FI20205405 A20211023FI2020000540520200422 WO21214393 A120211028WO2021FI5030120210421Link to current patent family on right

Drug delivery aims to optimize the systemic distribution of administered drugs to reach their target tissue in an effective and specific manner. This approach is arising also in the field of cancer therapy, since traditional small molecule chemotherapeutics can cause severe side effects and more sophisticated biomolecular therapeutics cannot always reach the target cells on their own. For these purposes, therapeutics can be packed into carriers that improve their pharmacokinetics by providing a suitable, protected environment for the cargo to travel in the body, armed with targeting molecules that guide them to the site of interest. Synthetic drug carriers include liposomes, porous silica and different polymer particles that can be modified to contain necessary surface structures for targeted delivery and improved biocompatibility. While they have shown much promise in research settings, their success in cancer treatment has been limited to only a handful of commercially available formulations, some of which have presented liver toxicity as a result of continuous administration. Additionally, targeting cancer cells is difficult because cancer is a very individual disease, and heterogeneity exists even within tumors, so finding any universal cancer-specific markers to target is extremely challenging. Nature has adopted a similar approach for delivering molecules containing biologically functional cargo from cell to cell. These naturally occurring carriers are called extracellular vesicles (EVs), in contrast to intracellular vesicles that mediate cargo trafficking inside of the cells. EVs are 50 – 1000 nm in diameter, consisting of a lipid bilayer with embedded membrane proteins that encloses water-soluble biological cargo within. Their exact composition and cargo vary from cell to cell and depending on the condition of the cells that produce them. Additionally, cells have multiple pathways for EV secretion and production. For these reasons EVs comprise a very heterogeneous population of vesicles in composition and they also have multiple biological functions that have not been completely clarified yet. EVs have been found in practically all body fluids and organisms that have been studied, taking part in the normal and pathological functions of the organism. For example, procoagulant EVs are found in blood and saliva that participate in hemostasis, while EVs secreted by cancer cells promote the survival, growth and metastasis of the tumor. They are able to affect cells via membrane interactions and by delivering functional cargo into their recipient cells. It has been shown that EVs are even able to target cells selectively by binding to specific cellular receptors, enabling targeted cargo delivery. Given their natural cargo delivery properties, EVs present remarkable potential for drug delivery applications. As cancer cells also use EVs to communicate with each other, their EVs reflect the same heterogeneity that exists within the cells themselves, and may provide useful insights for cancer-targeted drug delivery. However, the internalization mechanisms of drug-carrying EVs and especially the fate of the drugs they carry is not well understood. In this thesis, the use of EVs was assessed for the delivery of chemotherapeutic drug paclitaxel and an oncolytic adenovirus that represent small molecular and biological chemotherapeutics, with cancer cell-derived EVs as the model carrier. The studies presented here focus on the preparation, characterization, intracellular tracking and effectiveness of these EVs for cancer therapy. The obtained results provide novel methods and knowledge for the future development of EV-based therapeutics for the treatment of cancer. First, EVs were loaded with paclitaxel by incubation, which enhanced the cytotoxic effect of the drug, changing its internalization from passive diffusion to endocytosis. A novel approach using fluorescence lifetime microscopy was introduced for tracking the release of paclitaxel from the EVs inside of the cells, identifying distinct patterns of subcellular drug release in individual cells and its intracellular kinetics. This method was able to show details about the drug release mediated by EVs that could not be observed with conventional fluorescence microscopy. Additionally, the relationship between EVs and adenoviruses was explored, revealing a previously undocumented pathway of spreading adenoviral infection via EVs. This EV-mediated infective delivery of the viral genome occurred separately from the classical pathway of adenoviral life cycle, and produced infective particles resembling EVs more than virions. EVs can in theory enhance the pharmacokinetics of oncolytic viruses by hiding them from the immune system and providing them alternative pathways for cell targeting and internalization, however it was found that infective EVs do react with adenovirus neutralizing antibodies. Taken together, these results suggest that EVs can act as versatile carriers of therapeutic cargo for cancer treatment, ranging from small molecule chemotherapeutics to oncolytic viruses, though they require further development. New methods are reported constantly for preparing and studying therapeutic EVs with new, innovative approaches that will help in the future treatments of cancer and other diseases. Solunulkoiset vesikkelit (EV, Extracellular Vesicle) toimivat solujen välisinä viestinviejinä ja omaavat siksi huomattavia sovellutusmahdollisuuksia lääkkeenkuljetuksessa. Koska syöpäsolut myös käyttävät EV:itä kommunikoidakseen toistensa kanssa, niiden EV:t voivat tarjota hyödynnettävissä olevaa tietoa syöpään kohdennettavaan lääkkeenkuljetukseen. Lääkeaineita kantavien EV:iden sisäänottomekanismeja ja erityisesti lääkeaineiden kohtaloa ei kuitenkaan tunneta tarkasti. Tässä väitöskirjatyössä selvitettiin EV:iden käyttöä kemoterapiassa käytetyn paklitakselin ja onkolyyttisen adenoviruksen kuljetuttajina, jotka edustavat pienmolekulaarista ja biologista kemoterapiaa, käyttäen syöpäsolujen EV:itä mallikantajina. Tutkimuksissa keskityttiin näiden EV:iden valmistukseen, karakterisointiin, solunsisäiseen kulkeutumiseen ja toiminnallisuuteen syövän hoidossa. Saadut tulokset tarjoavat uusia menetelmiä ja tuntemusta tulevaisuuden EV-pohjaisten syöpähoitojen kehittämiseen. Ensin, EV:t lastattiin paklitakselilla inkuboimalla niitä yhdessä, mikä tehosti lääkeaineen sytotoksista vaikutusta, ohjaten sen sisäänottoa passiivisesta diffuusiosta endosytoosiin. Lisäksi esiteltiin uudenlaista lähestymistapaa paklitakselin EV:istä vapautumisen seuraamiseksi solujen sisällä fluoresenssin elinaikamikroskopialla, minkä avulla tunnistettiin lääkeaineen solunsisäisen vapautumisen mekanismeja yksittäisissä soluissa ja sen solunsisäistä kinetiikkaa. Lisäksi selvitettiin EV:iden ja adenovirusten välistä suhdetta, josta havaittiin aiemmin tunnistamaton reitti adenovirusinfektion levittämiseksi EV:illä. Tämä EV-välitteinen infektiivinen virusgenomin kuljetus ilmeni itsenäisenä adenoviruksen klassisesta elinkaaresta, infektiivisten partikkelien muistuttaen enemmän EV:itä kuin virioneja. Nämä tulokset viittaavat siihen, että EV:t voivat toimia monipuolisina kantajina syövän hoidossa käytettävälle terapeuttiselle lastille pienmolekyyleistä onkolyyttisiin viruksiin, tosin niiden käyttö edellyttää lisää kehitystyötä. Uusia menetelmiä esitetään jatkuvasti terapeuttisten EV:iden valmistamiseen ja tutkimukseen uusin, innovatiivisin lähestymistavoin, jotka tulevat edistämään tulevaisuudessa syövän ja muiden sairauksien hoitoa.

A microelectromechanical (MEMS) Fabry-Perot interferometer includes a transparent substrate; a first metallic mirror structure on the transparent substrate, including a first metal layer and a first support layer; a second metallic mirror structure above the first metallic mirror structure on an opposite side of the first metallic mirror structure in view of the transparent substrate, the second metallic mirror structure including a second metal layer and a second support layer, wherein the first and the second support layer are parallel and including at least one of aluminum oxide or titanium dioxide; a Fabry-Perot cavity between the first and the second support layer, whereby the Fabry-Perot cavity is formed by providing an insulation layer on the first mirror structure, and at least partially removing the insulation layer after providing the second mirror structure; and electrodes for providing electrical contacts to the first and the second metal layer.Patent family as of15.12.2021CN110809711 A 20200218 CN201880043636 20180629 EP3635351 A1 20200415 EP20180828785 20180629 EP3635351 A4 20200617 EP20180828785 20180629 FI128101 B 20190930 FI20170005641 20170703 FI20175641 A 20190104 FI20170005641 20170703 JP2020525830 T2 20200827 JP20190571313T 20180629 JP6870123 B2 20210512 JP20190571313T 20180629 US10732041 BB 20200804 US20190721975 20191220 US2020124475 AA 20200423 US20190721975 20191220 WO19008229 A1 20190110 WO2018FI50520 20180629Link to currentpatent family on right

An arrangement for hyperspectral imaging, comprising an imaging sensor (170,270); a band-pass filter element (130,230): at least one imaging optics element (120,160,220,260) configured to form an image on the imaging sensor (170,270); and a first adjustable multi passband filter (150a,255); wherein the first (150a,255) adjustable multi passband filter is configured to be adjusted by tilting.Patent family as of 10.12.2021CN108780008 A 20181109 CN201780017747 20170308 CN108780008 B 20210507 CN201780017747 20170308 EP3427023 A1 20190116 EP20170765914 20170308 EP3427023 A4 20191120 EP20170765914 20170308 FI126608 B 20170315 FI20160005213 20160315 FI20165213 A 20170315 FI20160005213 20160315 JP2019511870 T2 20190425 JP20180548218T 20170308 US2017269351 AA 20170921 US20170459164 20170315 US9835847 BB 20171205 US20170459164 20170315 WO17158231 A1 20170921 WO2017FI50155 20170308Link to current patent family on right

The invention relates to a system and a method for optical measurement of a target, wherein the target is illuminated, either actively illuminated, reflecting ambient light, or self illuminating, and a measurement radiation beam received from the target or through it is detected. The measurement system has optical fibres for guiding radiation from/to target positions. Radiation of several target positions is simultaneously filtered by a Fabry-Perot interferometer and detected by a row detector, for example.Patent family as of 21.10.2021US2014071451 AA 20140313 US20130024832 20130912 US9255844 BB 20160209 US20130024832 20130912Link to current patent family on right