Pub Date : 2023-10-25DOI: 10.18054/pb.v125i1-2.24860
Dominik Hamer, Daniela Petrinec, Monika Berecki, Anton Glasnović, Srećko Gajović
Recent advances in microscopy have enabled cellular-resolution imaging of thick tissue samples or even whole organs. The natural opacity of organs and tissues acts as a barrier to light penetration and must be removed to visualise structures of interest on a three-dimensional scale. Tissue optical clearing methods achieve sample transparency while also preserving fluorescently labelled epitopes. This innovative approach to sample preparation effectively enhances traditional histological sections and, with the aid of light sheet microscopy, enables optical sectioning and three-dimensional reconstruction of entire organs, even whole brains. Light sheet microscopy of optically cleared brain samples is a valuable method in neuroscience that is used in neuro-oncology, traumatic brain injury, ischemic brain injury, and neurodegenerative disease research.In this review, we describe tissue optical clearing methods used to achieve optical transparency in brain samples. This quickly developing field has a significant potential for producing cutting-edge uses in neuroscience research.
{"title":"Tissue optical clearing methods for microscopy: A review of their application in neuroscience","authors":"Dominik Hamer, Daniela Petrinec, Monika Berecki, Anton Glasnović, Srećko Gajović","doi":"10.18054/pb.v125i1-2.24860","DOIUrl":"https://doi.org/10.18054/pb.v125i1-2.24860","url":null,"abstract":"Recent advances in microscopy have enabled cellular-resolution imaging of thick tissue samples or even whole organs. The natural opacity of organs and tissues acts as a barrier to light penetration and must be removed to visualise structures of interest on a three-dimensional scale. Tissue optical clearing methods achieve sample transparency while also preserving fluorescently labelled epitopes. This innovative approach to sample preparation effectively enhances traditional histological sections and, with the aid of light sheet microscopy, enables optical sectioning and three-dimensional reconstruction of entire organs, even whole brains. Light sheet microscopy of optically cleared brain samples is a valuable method in neuroscience that is used in neuro-oncology, traumatic brain injury, ischemic brain injury, and neurodegenerative disease research.In this review, we describe tissue optical clearing methods used to achieve optical transparency in brain samples. This quickly developing field has a significant potential for producing cutting-edge uses in neuroscience research.","PeriodicalId":19950,"journal":{"name":"Periodicum Biologorum","volume":"8 3","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135218727","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-10-25DOI: 10.18054/pb.v125i1-2.25140
Vedrana Filić, Igor Weber
Three-dimensional imaging of fast intracellular processes by fluorescence microscopy should provide decent spatial and high temporal resolution while minimizing fluorophore bleaching and cytotoxicity. We give a condensed introductory overview of three contemporary methods mostly used for imaging of living cells in 3D and compare their performance in terms of temporal and spatial resolution, imaging flexibility and specimen photodamage: point-scanning confocal microscopy, spinning-disc confocal microscopy, and lattice light-sheet microscopy. While point-scanning instruments are unsurpassed in terms of confocal performance, flexibility and configurability of their optical path, spinning-disc and lattice light-sheet optical designs excel in acquisition speed and low levels of light-inflicted specimen deterioration.
{"title":"A young researcher’s guide to three-dimensional fluorescence microscopy of living cells","authors":"Vedrana Filić, Igor Weber","doi":"10.18054/pb.v125i1-2.25140","DOIUrl":"https://doi.org/10.18054/pb.v125i1-2.25140","url":null,"abstract":"Three-dimensional imaging of fast intracellular processes by fluorescence microscopy should provide decent spatial and high temporal resolution while minimizing fluorophore bleaching and cytotoxicity. We give a condensed introductory overview of three contemporary methods mostly used for imaging of living cells in 3D and compare their performance in terms of temporal and spatial resolution, imaging flexibility and specimen photodamage: point-scanning confocal microscopy, spinning-disc confocal microscopy, and lattice light-sheet microscopy. While point-scanning instruments are unsurpassed in terms of confocal performance, flexibility and configurability of their optical path, spinning-disc and lattice light-sheet optical designs excel in acquisition speed and low levels of light-inflicted specimen deterioration.","PeriodicalId":19950,"journal":{"name":"Periodicum Biologorum","volume":"9 5","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135218731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-10-25DOI: 10.18054/pb.v125i1-2.24852
Danijela Poljuha, Mirela Uzelac, Tea Zubin Ferri, Danijela Damijanić, Mihaela Šimunić, Hrvoje Korovljević, Tim Weber, Barbara Sladonja
Background and Purpose: Ailanthus altissima (Mill.) Swingle (Tree of Heaven) is a highly invasive, widespread, and widely investigated plant species native to the broader area of China. Extrafloral nectaries are major components of the A. altissima secretory system, but the knowledge of their morphology and role in the tree's physiology is limited. This research aims to explore the morphology of extrafloral nectaries, compare it to previous findings, and discuss their possible role and function. Materials and Methods: Extrafloral nectaries on leaves have been monitored through different phases of leaf development, from June to August 2015 and 2022. The nectaries' morphology was investigated using Zeiss Axioscope 5 and BOECO BSZ-405 light microscopes, and FEG QUANTA 250 FEI scanning electron microscope, operating at 7 kV and pressure of 60–100 Pa, without samples pretreatment. Results: Our investigation revealed the absence of earlier reported pores or ducts on the top of the glands. However, it supports one of the first, systematical investigations of A. altissima extrafloral nectaries conducted in Croatia a century ago, which was until recently forgotten by history. We evaluated our findings against prior theories and assessed the nectaries' potential role and purpose in disposing surplus sugars. Conclusions: The nectar in A. altissima is not secreted via an opening on the leaves but rather through epidermal tissue tearing.
{"title":"Morphology of extrafloral nectaries of Ailanthus altissima (Mill.) Swingle (Simaroubaceae)","authors":"Danijela Poljuha, Mirela Uzelac, Tea Zubin Ferri, Danijela Damijanić, Mihaela Šimunić, Hrvoje Korovljević, Tim Weber, Barbara Sladonja","doi":"10.18054/pb.v125i1-2.24852","DOIUrl":"https://doi.org/10.18054/pb.v125i1-2.24852","url":null,"abstract":"Background and Purpose: Ailanthus altissima (Mill.) Swingle (Tree of Heaven) is a highly invasive, widespread, and widely investigated plant species native to the broader area of China. Extrafloral nectaries are major components of the A. altissima secretory system, but the knowledge of their morphology and role in the tree's physiology is limited. This research aims to explore the morphology of extrafloral nectaries, compare it to previous findings, and discuss their possible role and function. Materials and Methods: Extrafloral nectaries on leaves have been monitored through different phases of leaf development, from June to August 2015 and 2022. The nectaries' morphology was investigated using Zeiss Axioscope 5 and BOECO BSZ-405 light microscopes, and FEG QUANTA 250 FEI scanning electron microscope, operating at 7 kV and pressure of 60–100 Pa, without samples pretreatment. Results: Our investigation revealed the absence of earlier reported pores or ducts on the top of the glands. However, it supports one of the first, systematical investigations of A. altissima extrafloral nectaries conducted in Croatia a century ago, which was until recently forgotten by history. We evaluated our findings against prior theories and assessed the nectaries' potential role and purpose in disposing surplus sugars. Conclusions: The nectar in A. altissima is not secreted via an opening on the leaves but rather through epidermal tissue tearing.","PeriodicalId":19950,"journal":{"name":"Periodicum Biologorum","volume":"11 7","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135218737","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-10-25DOI: 10.18054/pb.v125i1-2.23766
Ivana Restović, Marko Vučemilo, Mia Obad, Nives Kević, Nela Kelam, Anita Racetin, Ivana Bočina
Background and purpose: Dendrin is a brain and renal protein that is supposed to be involved in cytoskeletal modifications at the synapse and a part of the slit diaphragm and podocytes. Here, we aimed to investigate dendrin expression in dogfish brain since this newly discovered protein was never reported in fish. We compared the expression of dendrin to those of glial (GFAP) and neuronal (NF) proteins, which have already been described in the dogfish brain. Materials and methods: Histological and immunofluorescent techniques were performed on tissue samples. The obtained data were statistically analyzed. Results: Our results have shown that dendrin is expressed in all observed parts of the dogfish brain. In the forebrain, both observed parts (telencephalon and olfactory lobes) expressed dendrin. Regarding the percentage area of dendrin expression, it is expressed more in olfactory lobes than in the telencephalon. Compared with GFAP and NF expression, the expression of dendrin significantly differs in both parts of the forebrain. The highest dendrin expression was noticed in the midbrain. In dogfish midbrain, the difference in expression of dendrin in comparison to those of GFAP and NF was even more significant. The percentage area of dendrin expression in the hindbrain (cerebellum and medulla oblongata) was smaller than those in the forebrain and midbrain, contrary the percentage area of intermediate filaments GFAP and NF were significantly higher. Conclusion: These results are the first report on dendrin expression in the dogfish brain opening the path for future studies on its role and function.
{"title":"Expression of dendrin, neurofilament and glial fibrillary acidic protein in the brain of the dogfish Scyliorhinus canicula L.","authors":"Ivana Restović, Marko Vučemilo, Mia Obad, Nives Kević, Nela Kelam, Anita Racetin, Ivana Bočina","doi":"10.18054/pb.v125i1-2.23766","DOIUrl":"https://doi.org/10.18054/pb.v125i1-2.23766","url":null,"abstract":"Background and purpose: Dendrin is a brain and renal protein that is supposed to be involved in cytoskeletal modifications at the synapse and a part of the slit diaphragm and podocytes. Here, we aimed to investigate dendrin expression in dogfish brain since this newly discovered protein was never reported in fish. We compared the expression of dendrin to those of glial (GFAP) and neuronal (NF) proteins, which have already been described in the dogfish brain. Materials and methods: Histological and immunofluorescent techniques were performed on tissue samples. The obtained data were statistically analyzed. Results: Our results have shown that dendrin is expressed in all observed parts of the dogfish brain. In the forebrain, both observed parts (telencephalon and olfactory lobes) expressed dendrin. Regarding the percentage area of dendrin expression, it is expressed more in olfactory lobes than in the telencephalon. Compared with GFAP and NF expression, the expression of dendrin significantly differs in both parts of the forebrain. The highest dendrin expression was noticed in the midbrain. In dogfish midbrain, the difference in expression of dendrin in comparison to those of GFAP and NF was even more significant. The percentage area of dendrin expression in the hindbrain (cerebellum and medulla oblongata) was smaller than those in the forebrain and midbrain, contrary the percentage area of intermediate filaments GFAP and NF were significantly higher. Conclusion: These results are the first report on dendrin expression in the dogfish brain opening the path for future studies on its role and function.","PeriodicalId":19950,"journal":{"name":"Periodicum Biologorum","volume":"2 2","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135218734","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-10-25DOI: 10.18054/pb.v125i1-2.23747
Edi Rođak, Milorad Zjalić, Sara Cibok, Sven Viland, Marija Heffer, Robert Grgac, Tatjana Belovari, Nada Oršolić, Nikola Bijelić
Background and purpose: Since the times of Golgi and Cajal, impregnation with gold or silver has been used to visualize microscopic details of the nervous tissue. Although immunohistochemistry has largely replaced impregnation techniques, they are still used, and there is a growing interest in combining them with modern image analysis methods for quantitative studies in neuroscience. The aim of this research was to modify the gold chloride impregnation method published by Schmued to improve consistency of staining, to be adequate for digital image analysis. Materials and methods: Brains of 8 six-month-old female Wistar rats were fixed in 4% PFA, cryoprotected in sucrose and flash‑frozen in liquid nitrogen. Neighboring sets of coronal sections were chosen for gold impregnation, Nissl staining and MAP2 immunohistochemistry. Whole-slide images and images of specific regions were taken for analysis. Results: Myelinated fibers were stained dark reddish to brown on goldstained sections, and other tissue was yellowish, which gave an excellent contrast for digital image analysis. Gold staining was consistent in all regions, and no major artifacts were noticed. When compared to Nissl and MAP2, only myelinated structures were stained with gold impregnation. Conclusions: Modified gold impregnation method is an alternative that’s on par with traditional myelin staining methods. The new, modified gold impregnation method gives a consistent and reproducible staining suitable for digital image analysis. It can be useful in morphometric evaluation of nervous tissue and investigation of neuropathological changes in nervous tissue, especially for quantitative studies.
{"title":"Retro is the new modern: Contemporary application of gold impregnation staining on brain cryosections for digital image analysis","authors":"Edi Rođak, Milorad Zjalić, Sara Cibok, Sven Viland, Marija Heffer, Robert Grgac, Tatjana Belovari, Nada Oršolić, Nikola Bijelić","doi":"10.18054/pb.v125i1-2.23747","DOIUrl":"https://doi.org/10.18054/pb.v125i1-2.23747","url":null,"abstract":"Background and purpose: Since the times of Golgi and Cajal, impregnation with gold or silver has been used to visualize microscopic details of the nervous tissue. Although immunohistochemistry has largely replaced impregnation techniques, they are still used, and there is a growing interest in combining them with modern image analysis methods for quantitative studies in neuroscience. The aim of this research was to modify the gold chloride impregnation method published by Schmued to improve consistency of staining, to be adequate for digital image analysis. Materials and methods: Brains of 8 six-month-old female Wistar rats were fixed in 4% PFA, cryoprotected in sucrose and flash‑frozen in liquid nitrogen. Neighboring sets of coronal sections were chosen for gold impregnation, Nissl staining and MAP2 immunohistochemistry. Whole-slide images and images of specific regions were taken for analysis. Results: Myelinated fibers were stained dark reddish to brown on goldstained sections, and other tissue was yellowish, which gave an excellent contrast for digital image analysis. Gold staining was consistent in all regions, and no major artifacts were noticed. When compared to Nissl and MAP2, only myelinated structures were stained with gold impregnation. Conclusions: Modified gold impregnation method is an alternative that’s on par with traditional myelin staining methods. The new, modified gold impregnation method gives a consistent and reproducible staining suitable for digital image analysis. It can be useful in morphometric evaluation of nervous tissue and investigation of neuropathological changes in nervous tissue, especially for quantitative studies.","PeriodicalId":19950,"journal":{"name":"Periodicum Biologorum","volume":"42 4","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135218736","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-10-25DOI: 10.18054/pb.v125i1-2.25775
Miran Čeh, Jitka Hreščak, Sandra Drev
Modern state-of-the-art electron microscopy (EM) techniques are among the most versatile and useful analytical methods for nondestructive morphological, structural, and chemical characterization of materials. Both, scanning electron microscopy techniques (SEM) and transmission electron microscopy techniques (TEM), in essence complementary analytical methods, are capable of providing insight into the surface morphology, structure and chemical composition of materials from micrometer range to sub-atomic range. The importance of electron microscopy techniques in materials science have been readily recognized by the researchers of the Jozef Stefan Institute very early after its foundation in 1949 which resulted in the purchase of the first transmission electron microscopy Carl Zeiss EM-8 already in 1954. During the following years many researchers and institute directors have put a lot of effort into building and developing EM research infrastructure at the institute with accompanying expertise.
{"title":"SEM and TEM investigations of materials at the Centre for Electron Microscopy and Microanalysis (CEMM)","authors":"Miran Čeh, Jitka Hreščak, Sandra Drev","doi":"10.18054/pb.v125i1-2.25775","DOIUrl":"https://doi.org/10.18054/pb.v125i1-2.25775","url":null,"abstract":"Modern state-of-the-art electron microscopy (EM) techniques are among the most versatile and useful analytical methods for nondestructive morphological, structural, and chemical characterization of materials. Both, scanning electron microscopy techniques (SEM) and transmission electron microscopy techniques (TEM), in essence complementary analytical methods, are capable of providing insight into the surface morphology, structure and chemical composition of materials from micrometer range to sub-atomic range. The importance of electron microscopy techniques in materials science have been readily recognized by the researchers of the Jozef Stefan Institute very early after its foundation in 1949 which resulted in the purchase of the first transmission electron microscopy Carl Zeiss EM-8 already in 1954. During the following years many researchers and institute directors have put a lot of effort into building and developing EM research infrastructure at the institute with accompanying expertise.","PeriodicalId":19950,"journal":{"name":"Periodicum Biologorum","volume":"32 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135219705","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-10-25DOI: 10.18054/pb.v125i1-2.26468
Danijela Poljuha, Goran Kovačević
We are delighted to present a special issue of Periodicum biologorum,dedicated to the 30th anniversary of the Croatian Microscopy Societyas an independent Society and the 40th anniversary of its foundation.
{"title":"Celebrating 40 years of the Croatian Microscopy Society","authors":"Danijela Poljuha, Goran Kovačević","doi":"10.18054/pb.v125i1-2.26468","DOIUrl":"https://doi.org/10.18054/pb.v125i1-2.26468","url":null,"abstract":"We are delighted to present a special issue of Periodicum biologorum,dedicated to the 30th anniversary of the Croatian Microscopy Societyas an independent Society and the 40th anniversary of its foundation.","PeriodicalId":19950,"journal":{"name":"Periodicum Biologorum","volume":"32 3","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135219704","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-10-25DOI: 10.18054/pb.v125i1-2.24077
Damir Sirovina, Daniela Petrinec, Petra Tramontana, Goran Kovačević
Background and purpose: For many years, students’ interest in the field of natural sciences has declined, which consequently results in decreasing numbers of professional staff in the mentioned field. The aim of this study was to examine the attendance of extracurricular activities in the field of natural sciences by elementary school students (age 13 and 14) in Zagreb and whether the teachers' expectations are in line with the students' preferences for the content offered as part of extracurricular activities in biology. Material and methods: Using an anonymous online survey (Google Forms), data were collected on participation in extracurricular activities in the field of natural sciences and the attractiveness of the content of extracurricular activities. The respondents were 90 students in the 7th grade and 46 teachers of biology in Zagreb, 2020. Results: Participation in extracurricular activities in the field of natural sciences among the surveyed students was very low. Students were attracted to content that includes simple practical work in the field of biology, including microscopy, but the perception of their interest by teachers is not in accordance with the real interests of the students. Conclusion: The results of this research show that the share of the science field in extracurricular activities is low, and that the surveyed teachers are not aware of the students' wishes regarding the content offered in extracurricular activities. The conducted research can be a good basis for organizing and planning extracurricular activities in elementary school.
{"title":"Attendance of extracurricular activities in the field of natural sciences and the attractiveness of the content offered for extracurricular activities in biology in elementary schools","authors":"Damir Sirovina, Daniela Petrinec, Petra Tramontana, Goran Kovačević","doi":"10.18054/pb.v125i1-2.24077","DOIUrl":"https://doi.org/10.18054/pb.v125i1-2.24077","url":null,"abstract":"Background and purpose: For many years, students’ interest in the field of natural sciences has declined, which consequently results in decreasing numbers of professional staff in the mentioned field. The aim of this study was to examine the attendance of extracurricular activities in the field of natural sciences by elementary school students (age 13 and 14) in Zagreb and whether the teachers' expectations are in line with the students' preferences for the content offered as part of extracurricular activities in biology. Material and methods: Using an anonymous online survey (Google Forms), data were collected on participation in extracurricular activities in the field of natural sciences and the attractiveness of the content of extracurricular activities. The respondents were 90 students in the 7th grade and 46 teachers of biology in Zagreb, 2020. Results: Participation in extracurricular activities in the field of natural sciences among the surveyed students was very low. Students were attracted to content that includes simple practical work in the field of biology, including microscopy, but the perception of their interest by teachers is not in accordance with the real interests of the students. Conclusion: The results of this research show that the share of the science field in extracurricular activities is low, and that the surveyed teachers are not aware of the students' wishes regarding the content offered in extracurricular activities. The conducted research can be a good basis for organizing and planning extracurricular activities in elementary school.","PeriodicalId":19950,"journal":{"name":"Periodicum Biologorum","volume":"27 8","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135218728","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-10-25DOI: 10.18054/pb.v125i1-2.24080
Anja Sadžak, Lucija Mandić, Suzana Šegota, Vida Strasser
Background: To provide a fundamental understanding of the potential and use of atomic force microscopy (AFM) in medicine and the life sciences, this work presents a thorough description of imaging and non-imaging atomic force microscopy modes for characterizing model membranes and cells at the nanoscale. Methods: The imaging and non-imaging AFM modes are described with examples in terms of the characterization of topographic, morphological, and nanomechanical sample properties. Results: AFM imaging of supported lipid bilayers (SLBs) revealed the effects of temperature and medium composition on SLB topography in the gel and fluid phases, and on the bilayer thickness. Non-imaging AFM showed the strengthening of the SLB in both phases by the ion binding process.Imaging of neuronal and neuroblastoma cells with and without treatment revealed morphological changes in shape, volume, roughness, and Feret dimension. Non-imaging AFM showed the change in cell elasticity induced by the treatment with H2O2 with and without quercetin and by the treatment with copper and myricetin. The measurements of cells elasticity revealed a reorganization of the cytoskeleton and filament structures. Conclusions: Diverse applications of imaging and non-imaging AFM can provide important information about the underlying processes in biologically relevant systems. AFM, as a complementary technique to other biomedical methods, allows screening and monitoring of physiological changes at the nanoscale.
{"title":"Use of atomic force microscopy for characterization of model membranes and cells","authors":"Anja Sadžak, Lucija Mandić, Suzana Šegota, Vida Strasser","doi":"10.18054/pb.v125i1-2.24080","DOIUrl":"https://doi.org/10.18054/pb.v125i1-2.24080","url":null,"abstract":"Background: To provide a fundamental understanding of the potential and use of atomic force microscopy (AFM) in medicine and the life sciences, this work presents a thorough description of imaging and non-imaging atomic force microscopy modes for characterizing model membranes and cells at the nanoscale. Methods: The imaging and non-imaging AFM modes are described with examples in terms of the characterization of topographic, morphological, and nanomechanical sample properties. Results: AFM imaging of supported lipid bilayers (SLBs) revealed the effects of temperature and medium composition on SLB topography in the gel and fluid phases, and on the bilayer thickness. Non-imaging AFM showed the strengthening of the SLB in both phases by the ion binding process.Imaging of neuronal and neuroblastoma cells with and without treatment revealed morphological changes in shape, volume, roughness, and Feret dimension. Non-imaging AFM showed the change in cell elasticity induced by the treatment with H2O2 with and without quercetin and by the treatment with copper and myricetin. The measurements of cells elasticity revealed a reorganization of the cytoskeleton and filament structures. Conclusions: Diverse applications of imaging and non-imaging AFM can provide important information about the underlying processes in biologically relevant systems. AFM, as a complementary technique to other biomedical methods, allows screening and monitoring of physiological changes at the nanoscale.","PeriodicalId":19950,"journal":{"name":"Periodicum Biologorum","volume":"92 7","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135218735","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-10-25DOI: 10.18054/pb.v125i1-2.23764
Nives Kević, Ivana Restović, Ivana Bočina
Background and purpose: In the vertebrate family, the epidermis of terrestrial animals is keratinized while in aquatic forms and amphibians can produce mucus. Amphioxus, a cephalochordate, is an important animal model in the study of chordate phylogeny. Major cytoskeletal and extracellular matrix proteins in epidermal cells of amphioxus (Branchiostoma lanceolatum L.) were investigated using transmission electron microscopy and immunofluorescence. Materials and methods: Amphioxus specimens were fresh caught in the Adriatic Sea. Tissue samples were fixed and prepared for transmission electron microscopy and immunofluorescence techniques. Results: The epidermis consisted of one-layered columnar epithelium. Epithelial cells contained voluminous nucleus, a well-developed supranuclear Golgi apparatus, abundant vesicles and apical secretory vesicles surrounded by cytokeratin filaments. A single row of smaller vesicles also appeared close to the basal cell membrane. Strong positive immunolocalization of hyaluronic acid and collagen type I was observed in the apical domain of the plasma membrane of amphioxus, respectively. The positive signal of hyaluronan and collagen proteins referred to the secretory granules in the apical domain of the epidermal cell. Strong positive labelling of aggrecan was seen around the cell nucleus, deep below the apical domain, suggesting that aggrecan is not located in the secretory granules. A strong signal of the cytokeratin 10 protein was seen throughout the cytoplasm of the epidermal cells. Conclusion: Due to its morphology and histochemical composition, epidermal cells of amphioxus may contribute to course surface protection and the mucous layer in amphioxus may serve as a physical barrier between the body and its environment.
{"title":"Ultrastructural and immunofluorescence features of the epidermal cells and its secretory granules in the amphioxus Branchiostoma lanceolatum L.","authors":"Nives Kević, Ivana Restović, Ivana Bočina","doi":"10.18054/pb.v125i1-2.23764","DOIUrl":"https://doi.org/10.18054/pb.v125i1-2.23764","url":null,"abstract":"Background and purpose: In the vertebrate family, the epidermis of terrestrial animals is keratinized while in aquatic forms and amphibians can produce mucus. Amphioxus, a cephalochordate, is an important animal model in the study of chordate phylogeny. Major cytoskeletal and extracellular matrix proteins in epidermal cells of amphioxus (Branchiostoma lanceolatum L.) were investigated using transmission electron microscopy and immunofluorescence. Materials and methods: Amphioxus specimens were fresh caught in the Adriatic Sea. Tissue samples were fixed and prepared for transmission electron microscopy and immunofluorescence techniques. Results: The epidermis consisted of one-layered columnar epithelium. Epithelial cells contained voluminous nucleus, a well-developed supranuclear Golgi apparatus, abundant vesicles and apical secretory vesicles surrounded by cytokeratin filaments. A single row of smaller vesicles also appeared close to the basal cell membrane. Strong positive immunolocalization of hyaluronic acid and collagen type I was observed in the apical domain of the plasma membrane of amphioxus, respectively. The positive signal of hyaluronan and collagen proteins referred to the secretory granules in the apical domain of the epidermal cell. Strong positive labelling of aggrecan was seen around the cell nucleus, deep below the apical domain, suggesting that aggrecan is not located in the secretory granules. A strong signal of the cytokeratin 10 protein was seen throughout the cytoplasm of the epidermal cells. Conclusion: Due to its morphology and histochemical composition, epidermal cells of amphioxus may contribute to course surface protection and the mucous layer in amphioxus may serve as a physical barrier between the body and its environment.","PeriodicalId":19950,"journal":{"name":"Periodicum Biologorum","volume":"21 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135218729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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