Developing correlative Optical-Photoacustic microscopy for high-resolution in vivo imaging

PhotoAcoustic Imaging (PAI) is a branch in clinical and pre-clinical imaging, that refers to the techniques mapping acoustic signals caused by the absorption of the short laser pulse. This conversion of electromagnetic energy of the light to the mechanical (acoustic) energy is usually called photoacoustic effect. PAI, by combining optical excitation with acoustical detection, is able to preserve the diffraction limited spatial resolution. At the same time, the penetration depth is extended beyond the diffusive limit. The Laser-Scanning PhotoAcoustic Microscope system (LS-PAM) has been developed, that offers the axial resolution of 7.75 µm with the lateral resolution better than 10 µm. The first in vivo imaging experiments were carried out. Thus, in vivo label-free imaging of the mouse ear was performed. The principle possibility to image vessels located in deep layers of the mouse skin was shown. As well as that, a gold printing sample, vasculature of the Chick Chorioallantoic Membrane Assay, Drosophila larvae were imaged by PAI. During the experimental work, a totally new application of PAM was found, in which the acoustic waves, generated by incident light can be used for further imaging of another sample. In order to enhance the performance of the presented system two main recommendation can be offered. First, the current system should be transformed into reflection-mode setup system. Second, a more powerful source of light with the sufficient repetition rate should be introduced into the system.

European Association of Veterinary Diagnostic Imaging : 4th European conferece : matkakertomus

Mirja Ruohoniemi

Real-time Imaging and Mosaicking of Planar Surfaces

Laajojen pintojen kuvaaminen rajoitetussa työskentelytilassa riittävällä kuvatarkkuudella voi olla vaikeaa. Kuvaaminen on suoritettava osissa ja osat koottava saumattomaksi kokonaisnäkymäksi eli mosaiikkikuvaksi. Kuvauslaitetta käsin siirtelevän käyttäjän on saatava välitöntä palautetta, jotta mosaiikkiin ei jäisi aukkoja ja työ olisi nopeaa. Työn tarkoituksena oli rakentaa pieni, kannettava ja tarkka kuvauslaite paperi- ja painoteollisuuden tarpeisiin sekä kehittää palautteen antamiseen menetelmä, joka koostaaja esittää karkeaa mosaiikkikuvaa tosiajassa. Työssä rakennettiin kaksi kuvauslaitetta: ensimmäinen kuluttajille ja toinen teollisuuteen tarkoitetuista osista. Kuvamateriaali käsiteltiin tavallisella pöytätietokoneella. Videokuvien välinen liike laskettiin yksinkertaisella seurantamenetelmällä ja mosaiikkikuvaa koottiin kameroiden kuvanopeudella. Laskennallista valaistuksenkorjausta tutkittiin ja kehitetty menetelmä otettiin käyttöön. Ensimmäisessä kuvauslaitteessa on ongelmia valaistuksen ja linssivääristymien kanssa tuottaen huonolaatuisia mosaiikkikuvia. Toisessa kuvauslaitteessa nämä ongelmat on korjattu. Seurantamenetelmä toimii hyvin ottaen huomioon sen yksinkertaisuuden ja siihen ehdotetaan monia parannuksia. Työn tulokset osoittavat, että tosiaikainen mosaiikkikuvan koostaminen megapikselin kuvamateriaalista on mahdollista kuluttajille tarkoitetulla tietokonelaitteistolla.

Development of the Acoustic Emission Based Diagnostics Concept for the Rotary Lime Kiln

Reaaliaikainen, ennakoiva kunnonvalvonta on erittäin tärkeä osa modernin tehtaan tai tuotantolinjan toimintaa. Diplomityön teettäjä haluaa edelleen kehittää akustiseen emissioon perustuvaa kunnonvalvonta järjestelmäänsä, jotta siitä olisi enemmän hyötyä asiakkaalle. Diplomityö sisältää johdannonakustiseen emissioon ja akustisiin emissio sensoreihin. Työn tavoitteena oli kehittää päätöksentekojärjestelmä, jota käytettäisiin työn teettäjän valmistamien sensoreiden antaman tiedon automatisoituun analysointiin. Työssä on vertailtu kolmea eri ohjelmistotoimittajaa ja heidän ohjelmiaan, ja tehty ehdotus hankittavasta ohjelmistosta. Lisäksi työssä on kehitetty ohjeita, joiden avulla ohjelmisto ohjelmoidaan tuottamaan reaaliaikaista tietoa ja huolto-ohjeita sen käyttäjille. Lisäksi työssä annetaan ehdotuksia kunnonvalvonta- ja päätöksentekojärjestelmän edelleen kehittämiseen.

Performance Evaluation of Imaging Systems

Imaging systems have developed latest years and developing is still continuing following years. Manufacturers of imaging systems give promises for the quality of the performance of imaging systems to advertise their products. Promises for the quality of the performance are often so good that they will not be tested in normal usage. The main target in this research is to evaluate the quality of the performance of two imaging systems: Scanner and CCD color camera. Optical measurement procedures were planned to evaluate the quality of imaging performances. Other target in this research is to evaluate calibration programs for the camera and the scanner. Measuring targets had to choose to evaluate the quality of imaging performances. Manufacturers have given definitions for targets. The third task in this research is to evaluate and consider how good measuring targets are.

Imaging techniques applied for detection of secretion, transgene delivery and G proteincoupled receptors in reproductive and endocrine cells in vivo and in vitro

Post-testicular sperm maturation occurs in the epididymis. The ion concentration and proteins secreted into the epididymal lumen, together with testicular factors, are believed to be responsible for the maturation of spermatozoa. Disruption of the maturation of spermatozoa in the epididymis provides a promising strategy for generating a male contraceptive. However, little is known about the proteins involved. For drug development, it is also essential to have tools to study the function of these proteins in vitro. One approach for screening novel targets is to study the secretory products of the epididymis or the G protein-coupled receptors (GPCRs) that are involved in the maturation process of the spermatozoa. The modified Ca2+ imaging technique to monitor release from PC12 pheochromocytoma cells can also be applied to monitor secretory products involved in the maturational processes of spermatozoa. PC12 pheochromocytoma cells were chosen for evaluation of this technique as they release catecholamines from their cell body, thus behaving like endocrine secretory cells. The results of the study demonstrate that depolarisation of nerve growth factor -differentiated PC12 cells releases factors which activate nearby randomly distributed HEL erythroleukemia cells. Thus, during the release process, the ligands reach concentrations high enough to activate receptors even in cells some distance from the release site. This suggests that communication between randomly dispersed cells is possible even if the actual quantities of transmitter released are extremely small. The development of a novel method to analyse GPCR-dependent Ca2+ signalling in living slices of mouse caput epididymis is an additional tool for screening for drug targets. By this technique it was possible to analyse functional GPCRs in the epithelial cells of the ductus epididymis. The results revealed that, both P2X- and P2Y-type purinergic receptors are responsible for the rapid and transient Ca2+ signal detected in the epithelial cells of caput epididymides. Immunohistochemical and reverse transcriptase-polymerase chain reaction (RTPCR) analyses showed the expression of at least P2X1, P2X2, P2X4 and P2X7, and P2Y1 and P2Y2 receptors in the epididymis. Searching for epididymis-specific promoters for transgene delivery into the epididymis is of key importance for the development of specific models for drug development. We used EGFP as the reporter gene to identify proper promoters to deliver transgenes into the epithelial cells of the mouse epididymis in vivo. Our results revealed that the 5.0 kb murine Glutathione peroxidase 5 (GPX5) promoter can be used to target transgene expression into the epididymis while the 3.8 kb Cysteine-rich secretory protein-1 (CRISP-1) promoter can be used to target transgene expression into the testis. Although the visualisation of EGFP in living cells in culture usually poses few problems, the detection of EGFP in tissue sections can be more difficult because soluble EGFP molecules can be lost if the cell membrane is damaged by freezing, sectioning, or permeabilisation. Furthermore, the fluorescence of EGFP is dependent on its conformation. Therefore, fixation protocols that immobilise EGFP may also destroy its usefulness as a fluorescent reporter. We therefore developed a novel tissue preparation and preservation techniques for EGFP. In addition, fluorescence spectrophotometry with epididymal epithelial cells in suspension revealed the expression of functional purinergic, adrenergic, cholinergic and bradykinin receptors in these cell lines (mE-Cap27 and mE-Cap28). In conclusion, we developed new tools for studying the role of the epididymis in sperm maturation. We developed a new technique to analyse GPCR dependent Ca2+ signalling in living slices of mouse caput epididymis. In addition, we improved the method of detecting reporter gene expression. Furthermore, we characterised two epididymis-specific gene promoters, analysed the expression of GPCRs in epididymal epithelial cells and developed a novel technique for measurement of secretion from cells.

Brain imaging studies in severe somatization

Somatization was described 4000 years ago but the pathophysiology of the, phenomenon is unknown. The aim of this investigation was to explore whether central nervous system (CNS) pathology is associated with severe somatization which was operationalized as somatization disorder (SD) and undifferentiated somatoform disorder. The study sample consisted of severely somatizing people who were included into the study after a multi-phase screening procedure in order to exclude psychiatric comorbidities and physical illnesses. Diagnosis of somatization disorder or undifferentiated sofatoform disorder were set according to Diagnostic and Statistical Manual of Mental Disorders 4th ed. (DSM-IV). The first study explored the regional cerebral metabolic rate of glucose (rCMRGlc) in severely somatizing females and found it to be reduced in several regions of the brain compared to healthy controls. The second study observed brain morphology with magnetic resonance imaging (MRI) based on the findings from the first study and showed enlarged caudate nuclei in somatizing women compared to healthy volunteers. The third study investigated temperament factors and brain metabolism, and their association with severe somatization. Low caudate and putamen metabolism, low novelty seeking as well as high harm avoidance were found to be associated with severe somatization in women, reduced caudate metabolism having the strongest association. The last study is a report of man with left-side gradient of multiple symptoms of unknown origin in the body. The examination revealed a hypermetabolic nucleus putamen on the contralateral side. All the main results reported in these four articles are original findings. The results suggest that CNS pathology is involved in the pathophysiology of severe somatization.