Individual Organelles Research Articles

Adaptive optimization of the OXPHOS assembly line partially compensates lrpprc-dependent mitochondrial translation defects in mice

Mouse models of genetic mitochondrial disorders are generally used to understand specific molecular defects and their biochemical consequences, but rarely to map compensatory changes allowing survival. Here we took advantage of the extraordinary mitochondrial resilience of hepatic Lrpprc knockout mice to explore this question using native proteomics profiling and lipidomics. In these mice, low levels of the mtRNA binding protein LRPPRC induce a global mitochondrial translation defect and a severe reduction (>80%) in the assembly and activity of the electron transport chain (ETC) complex IV (CIV). Yet, animals show no signs of overt liver failure and capacity of the ETC is preserved. Beyond stimulation of mitochondrial biogenesis, results show that the abundance of mitoribosomes per unit of mitochondria is increased and proteostatic mechanisms are induced in presence of low LRPPRC levels to preserve a balance in the availability of mitochondrial- vs nuclear-encoded ETC subunits. At the level of individual organelles, a stabilization of residual CIV in supercomplexes (SCs) is observed, pointing to a role of these supramolecular arrangements in preserving ETC function. While the SC assembly factor COX7A2L could not contribute to the stabilization of CIV, important changes in membrane glycerophospholipid (GPL), most notably an increase in SC-stabilizing cardiolipins species (CLs), were observed along with an increased abundance of other supramolecular assemblies known to be stabilized by, and/or participate in CL metabolism. Together these data reveal a complex in vivo network of molecular adjustments involved in preserving mitochondrial integrity in energy consuming organs facing OXPHOS defects, which could be therapeutically exploited.

The structure and function of centriolar rootlets.

ABSTRACTTo gain a holistic understanding of cellular function, we must understand not just the role of individual organelles, but also how multiple macromolecular assemblies function collectively. Centrioles produce fundamental cellular processes through their ability to organise cytoskeletal fibres. In addition to nucleating microtubules, centrioles form lesser-known polymers, termed rootlets. Rootlets were identified over a 100 years ago and have been documented morphologically since by electron microscopy in different eukaryotic organisms. Rootlet-knockout animals have been created in various systems, providing insight into their physiological functions. However, the precise structure and function of rootlets is still enigmatic. Here, I consider common themes of rootlet function and assembly across diverse cellular systems. I suggest that the capability of rootlets to form physical links from centrioles to other cellular structures is a general principle unifying their functions in diverse cells and serves as an example of how cellular function arises from collective organellar activity.

Illuminating the cells: transient transformation of citrus to study gene functions and organelle activities related to fruit quality

Although multiple microscopic techniques have been applied to horticultural research, few studies of individual organelles in living fruit cells have been reported to date. In this paper, we established an efficient system for the transient transformation of citrus fruits using an Agrobacterium-mediated method. Kumquat (Fortunella crassifolia Swingle) was used; it exhibits higher transformation efficiency than all citrus fruits that have been tested and a prolonged-expression window. Fruits were transformed with fluorescent reporters, and confocal microscopy and live-cell imaging were used to study their localization and dynamics. Moreover, various pH sensors targeting different subcellular compartments were expressed, and the local pH environments in cells from different plant tissues were compared. The results indicated that vacuoles are most likely the main organelles that contribute to the low pH of citrus fruits. In summary, our method is effective for studying various membrane trafficking events, protein localization, and cell physiology in fruit and can provide new insight into fruit biology research.

Reactive oxygen species and organellar signaling

The evolution of photosynthesis and its associated metabolic pathways has been crucial to the successful establishment of plants, but has also challenged plant cells in the form of production of reactive oxygen species (ROS). Intriguingly, multiple forms of ROS are generated in virtually every plant cell compartment through diverse pathways. As a result, a sophisticated network of ROS detoxification and signaling that is simultaneously tailored to individual organelles and safeguards the entire cell is necessary. Here we take an organelle-centric view on the principal sources and sinks of ROS across the plant cell and provide insights into the ROS-induced organelle to nucleus retrograde signaling pathways needed for operational readjustments during environmental stresses.

Network analysis of Arabidopsis mitochondrial dynamics reveals a resolved tradeoff between physical distribution and social connectivity.

SummaryMitochondria in plant cells exist largely as individual organelles which move, colocalize, and interact, but the cellular priorities addressed by these dynamics remain incompletely understood. Here, we elucidate these principles by studying the dynamic "social networks" of mitochondria in Arabidopsis thaliana wildtype and mutants, describing the colocalization of individuals over time. We combine single-cell live imaging of hypocotyl mitochondrial dynamics with individual-based modeling and network analysis. We identify an inevitable tradeoff between mitochondrial physical priorities (an even cellular distribution of mitochondria) and “social” priorities (individuals interacting, to facilitate the exchange of chemicals and information). This tradeoff results in a tension between maintaining mitochondrial spacing and facilitating colocalization. We find that plant cells resolve this tension to favor efficient networks with high potential for exchanging contents. We suggest that this combination of physical modeling coupled to experimental data through network analysis can shed light on the fundamental principles underlying these complex organelle dynamics. A record of this paper’s transparent peer review process is included in the supplemental information.

Subcellular Specialization of 3D Mitochondrial Morphology and Mitochondria‐Lipid Droplet Interactivity is Associated with Proximity to Oxygen Supply in Muscle Cells

Advancements in imaging technology have considerably expanded our capacity to investigate cell biology, and our laboratory has recently employed high-resolution 3D microscopy to explore the subcellular structures and organelle-organelle interactions that support muscle metabolism. While this initial work has demonstrated that mitochondrial structure and mitochondria-organelle interactivity are regulated according to muscle cell type, it remains unclear how cellular metabolism is supported by the subcellular specialization of mitochondria. Certainly, several subcellular clusters of mitochondria have been identified within muscle cells, and these groups of mitochondria are often categorized according to their proximity to other cellular components (i.e. “intermyofibrillar” or “paravascular”). However, it is unknown how 3D mitochondrial morphology and organelle interactivity is altered to support the varying functional demands of different subcellular regions. Here, we combine 3D focused-ion beam scanning electron microscopy (FIB-SEM) with machine learning programs to quantify individual mitochondrial structure and organelle interactivity within large subcellular volumes (~8,000 µm3) (Fig. 1a-c). Using this high-throughput approach, we evaluate over 700 individual mitochondria per cell and leverage the statistical power of this large data set to identify 3D spatial relationships among multiple subcellular structures. We find that mitochondrial surface area to volume ratio (15.5±0.25 µm-1) is significantly correlated (R2= 0.39) to the minimal distance to capillary, but not distance to sarcolemma (R2= 0.02) (Fig. 1d). Further analysis revealed that mitochondria-organelle interactivity is also closely related to subcellular localization. Specifically, mitochondria within close proximity (<5 µm) to capillaries (paravascular) are ~3-fold more likely to make contact with lipid droplets compared to mitochondria located in other subcellular regions (Fig. 1e). Moreover, the paravascular mitochondria in contact with lipid droplets have 75% greater volume (1.4±0.07 µm3) compared to those not connected to lipid droplets (0.8±0.03 µm3). These preliminary analyses suggest that mitochondria in close proximity to the muscle oxygen supply, but not cell boundary, exhibit distinct 3D morphology and organelle interactivity patterns that may function to support metabolism and other cellular processes.

Imaging the electrical activity of organelles in living cells

Eukaryotic cells are complex systems compartmentalized in membrane-bound organelles. Visualization of organellar electrical activity in living cells requires both a suitable reporter and non-invasive imaging at high spatiotemporal resolution. Here we present hVoSorg, an optical method to monitor changes in the membrane potential of subcellular membranes. This method takes advantage of a FRET pair consisting of a membrane-bound voltage-insensitive fluorescent donor and a non-fluorescent voltage-dependent acceptor that rapidly moves across the membrane in response to changes in polarity. Compared to the currently available techniques, hVoSorg has advantages including simple and precise subcellular targeting, the ability to record from individual organelles, and the potential for optical multiplexing of organellar activity.

Multiplex Raman imaging of organelles in endothelial cells

Raman imaging using molecular reporters is a relatively new approach in subcellular investigations. It enables the visualization of organelles in cells with better selectivity and sensitivity compared to the label-free approach. Essentially Raman reporters possess in their structure an alkyne molecular group that can be selectively identified in the spectral region silent for biomolecules, hence facilitate the localization of individual organelles. The aim of this work is to visualize the main cell organelles in endothelial cells (HMEC-1) using established reporters (EdU and MitoBADY), but also to test a new one, namely falcarinol, which exhibits lipophilic properties. Moreover, we tested the possibility to use Raman reporters as a probe to detect changes in distribution of certain organelles after induced endothelial dysfunction (ED) in in vitro models. In both cases, induced ED is characterized by the formation of lipid droplets in the cells, which is why a good tool for the detection of lipid-rich organelles is so important in these studies. Two-dimensional Raman images were obtained, visualizing the distribution of selected organic compounds in the cell, such as proteins, lipids, and nucleic acids. Additionally, the distribution of EdU, MitoBADY and falcarinol in endothelial cells (ECs) was determined. Moreover, we highlight some drawback of established Raman reporter and the need for testing them in various physiological state of the cell.

Layilin promotes mitochondrial fission by cyclin-dependent kinase 1 and dynamin-related protein 1 activation in HEK293T cells

ObjectFunctions of layilin, a type 1 transmembrane protein with a C-type lectin motif, remain to be clarified. We here investigated precise intracellular localization of layilin and the location-related functions. MethodsWe used HEK293T cells to assess the co-localization of layilin with different individual organelle markers by double immunostaining. We then investigated mitochondrial morphology in layilin-knockdown (KD) conditions, also with immunostaining. Next, we measured amounts of proteins involved in regulation of mitochondrial dynamics, DRP1, pS616-DRP1, mitofusin1, mitofusin2, CDK1, pY15-CDK1, and cyclin B1, in layilin-KD cells versus control cells by Western blot. Furthermore, by using layilin-knockout (KO) cells, amounts of CDK1 and pY15-CDK1 as well as mitochondrial morphology were investigated. ResultWe found that layilin localized to mitochondria rather than the other organelles. Small round-shape mitochondria were observed in control cells, whereas elongated and highly connected mitochondria were observed in layilin-KD cells. Amounts of active DRP1 (pS616-DRP1) and total DRP1 were significantly smaller in layilin-KD cells than in controls. Amounts of inactive CDK1 (pY15-CDK1) were significantly larger in layilin-KD cells than in controls. No other tested molecules were significantly altered in layilin-KD cells. Amounts of inactive CDK1 were significantly larger in layilin-KO cells than in wild type (WT) cells. Small round-shape mitochondria were observed in WT cells, whereas elongated and highly connected mitochondria were observed in layilin-KO cells. ConclusionWe here demonstrated that layilin played a role in the maintenance of fragmented mitochondria in mitochondrial dynamics and that this function needed CDK1 and DRP1 activation. Our data unveiled a novel function for layilin, regulation of mitochondrial dynamics.

Disease in a Dish Crispr-Edited HIPSC-CMS to Study the Cellular and Subcellular Mechanisms of HCM

Genetic sequencing enables the pinpointing of specific genetic mutations correlated with disease. However, such associations do not describe mechanistic relationships between specific mutations and clinical phenotypes. To address this, we use human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) as a model system to probe how mutations in β-myosin heavy chain (MHC) lead to disease. β-MHC, a major biochemical motor protein in heart muscle, is a common site of mutations implicated in hypertrophic cardiomyopathy Utilizing CRISPR editing of the MYH7 gene to introduce the G256E point mutation into the converter domain region of myosin, we have studied the effects that molecular level changes (nm) in protein structure have at the cellular level through simultaneous contractility and calcium handling measurements in hiPSC-CMs. These hiPSC-CMs were subjected to glucose starvation to mature and enrich the population of cardiomyocytes, and cultured on Matrigel-stamped 1500 μm2 island patterns to achieve single cells. We have also analyzed differences at the individual contractile organelle level through precise measures of myofibril contraction and relaxation kinetics using a custom-built apparatus. Our experimental results have been supplemented with molecular dynamics simulations of wild type and mutant β-MHC. Preliminary data from molecular dynamics simulations of β-myosin structure, suggests G256E alters the distributions of intra-molecular contacts made by loop 1 and distorts β-MHC structure near the nucleotide binding pocket as well as sites forming the interacting heads motif. Synthesis of computational, mechanical and structural studies will determine the mechanism of contractile dysfunction associated with β-MHC G256E. Development of these platforms for molecular and cellular level analysis serve as a template for future studies of other myofilament proteins and mutations to connect disease genotype with phenotype.

Targeted Solvatochromic Fluorescent Probes for Imaging Lipid Order in Organelles under Oxidative and Mechanical Stress.

Biomembranes constitute a basis for all compartments of live cells, and therefore, the monitoring of their lipid organization is essential for understanding cell status and activity. However, the sensing and imaging of lipid organization specifically in different organelles of live cells remain challenging. Here, we designed an array of solvatochromic probes based on Nile Red bearing ligands for specific targeting of the endoplasmic reticulum, mitochondria, lysosomes, Golgi apparatus, plasma membranes, and lipid droplets. These polarity-sensitive probes detected variations in the lipid order by changing their emission maximum, as evidenced by fluorescence spectroscopy in model membranes. In colocalization microscopy experiments with reference organelle markers, they exhibited good organelle selectivity. Using two-color fluorescence microscopy, the new probes enabled imaging of the local polarity of organelles in live cells. To exclude the biased effect of the probe design on the sensitivity to the membrane properties, we calibrated all probes in model membranes under the microscope, which enabled the first quantitative description of the lipid order in each organelle of interest. Cholesterol extraction/enrichment confirmed the capacity of the probes to sense the lipid order, revealing that organelles poor in cholesterol are particularly affected by its enrichment. The probes also revealed that oxidative and mechanical stresses produced changes in the local polarity and lipid order that were characteristic for each organelle, with mitochondria and lysosomes being particularly stress sensitive. The new probes constitute a powerful toolbox for monitoring the response of the cells to physical and chemical stimuli at the level of membranes of individual organelles, which remains an underexplored direction in cellular research.

Применение метода экспансионной микроскопии в нейробиологии

Many biological studies require the analysis of ultrastructural changes at the cellular organelles and molecular interaction. Since the modern confocal microscope resolution is limited by the diffraction limit (200-300nm), it is impossible to study such small objects using standard fluorescence microscopy. Ultra-high resolution microscopy methods require expensive equipment and are technically difficult in use, which in turn limits their widespread practical application. However, recently appeared methods make it possible to increase the resolution of microscopy not by improving the image registration system, but by physically isotropic expansion of a biological sample using a controlled chemical process. Thereby to this method, called expansion or expansive microscopy (EM), it became possible to obtain three-dimensional images of samples with a resolution sufficient for studying individual cell organelles using a conventional confocal microscope. This review covers the history of this method, its basic principles and examples of use in various fields of biology and medicine, as well as reflects future directions for improving this technology. The article discusses the methodological features of the EM applying in the study of brain tissue samples and primary neuronal cell culture with an algorithm that can be used to adapt the standard protocol to the goals and objectives of a particular study.

Innate immunity in C. elegans.

In its natural habitat, C. elegans encounters a wide variety of microbes, including food, commensals and pathogens. To be able to survive long enough to reproduce, C. elegans has developed a complex array of responses to pathogens. These activities are coordinated on scales that range from individual organelles to the entire organism. Often, the response is triggered within cells, by detection of infection-induced damage, mainly in the intestine or epidermis. C. elegans has, however, a capacity for cell non-autonomous regulation of these responses. This frequently involves the nervous system, integrating pathogen recognition, altering host biology and governing avoidance behavior. Although there are significant differences with the immune system of mammals, some mechanisms used to limit pathogenesis show remarkable phylogenetic conservation. The past 20 years have witnessed an explosion of host-pathogen interaction studies using C. elegans as a model. This review will discuss the broad themes that have emerged and highlight areas that remain to be fully explored.

Single organelle measurements of melanosome pH using the novel ratiometric indicator RpHiMEL.

Melanocytes are specialized cells that produce melanin pigments responsible for skin, hair, and eye pigmentation. The synthesis and storage of melanin occurs in unique lysosome-related organelles called melanosomes, which regulate melanin production via complex regulatory mechanisms. Maintenance of the melanosome luminal ionic environment and pH is crucial for proper function of the main melanogenic enzymes. Defects in genes encoding pH-regulating melanosomal proteins result in oculocutaneous albinism, which is characterized by hypopigmentation, impaired vision, and increased susceptibility to skin and eye cancers. We recently uncovered several ion channels and transporters that modulate melanin synthesis by acidifying or neutralizing the luminal pH of melanosomes. However, our understanding of how melanosomes and other related organelles maintain their luminal pH is far from complete. The study of melanosome pH regulation requires robust imaging and quantification tools. Despite recent advances in the development of such methods, many limitations remain, particularly for quantitative analysis of individual organelle pH. In this chapter, we will provide an overview of the available methods used for melanosome pH determination, including their advantages, limitations, and challenges. To address the critical, unmet need for reliable melanosome pH quantification tools, we engineered a novel genetically encoded, ratiometric pH sensor for melanosomes that we named RpHiMEL. Here, we describe the design and optimization of RpHiMEL, and provide a pH quantification method for individual melanosomes in live cells. We demonstrate that RpHiMEL is a highly versatile tool with the potential to advance our understanding of pH regulation in melanosomes and related organelles.

Study of the effect of natural polyphenols&#x0D; on the prostate cancer Du145 cell cycle

Relevance: Although natural polyphenols are known as&#x0D; useful nutrients, their use is quite occasional. This is probably&#x0D; due to a lack of complete information on their biochemical&#x0D; structure and mechanism of action. Scientific literature offers&#x0D; many studies on polyphenols’ benefits and metabolic effects&#x0D; on the body. Polyphenols are known to improve body function&#x0D; and increase physiological activity. Polyphenols are used in&#x0D; the body’s normal physiological function and metabolic disorders, and their therapeutic effect can be controlled in complex&#x0D; pathological conditions.&#x0D; Therefore, in our study, we used curcumin and carnosic&#x0D; acid to control the natural polyphenols’ metabolic effects’&#x0D; structure and function in prostate cancer. Metabolic disorders&#x0D; in mitochondria, one of the organelles responsible for cell energy metabolism, have an irreversible effect on the cell, organ,&#x0D; and the system as a whole. This urges the study of cellular action on individual organelles, including the biochemical and&#x0D; physiological aspects.&#x0D; The purpose was to determine the effect of natural polyphenols – curcumin and carnosic acid – on the prostate cancer&#x0D; cell cycle.&#x0D; Results: Combination of curcumin and carnosic acid caused&#x0D; an exposure time-dependent suppression of Du145 cancer cells&#x0D; growth at G0/1 and S phases.&#x0D; Conclusions: The effect of natural polyphenols – curcumin&#x0D; and carnosic acid – on the Du145 prostate cancer cell cycle was&#x0D; determined. Curcumin 7μM and carnosic acid 10μM in combination stopped phases G0/1 and S’ prostate cancer cells division

Micro- and Nano-Devices for Studying Subcellular Biology.

Cells are complex machines whose behaviors arise from their internal collection of dynamically interacting organelles, supramolecular complexes, and cytoplasmic chemicals. The current understanding of the nature by which subcellular biology produces cell-level behaviors is limited by the technological hurdle of measuring the large number (>103 ) of small-sized (<1μm) heterogeneous organelles and subcellular structures found within each cell. In this review, the emergence of a suite of micro- and nano-technologies for studying intracellular biology on the scale of organelles is described. Devices that use microfluidic and microelectronic components for 1) extracting and isolating subcellular structures from cells and lysate; 2) analyzing the physiology of individual organelles; and 3) recreating subcellular assembly and functions in vitro, are described. The authors envision that the continued development of single organelle technologies and analyses will serve as a foundation for organelle systems biology and will allow new insight into fundamental and clinically relevant biological questions.

SynapsEM: Computer-Assisted Synapse Morphometry

The structural features of a synapse help determine its function. Synapses are extremely small and tightly packed with vesicles and other organelles. Visualizing synaptic structure requires imaging by electron microscopy, and the features in micrographs must be quantified, a process called morphometry. Three parameters are typically assessed from each specimen: (1) the sizes of individual vesicles and organelles; (2) the absolute number and densities of organelles; and (3) distances between organelles and key features at synapses, such as active zone membranes and dense projections. For data to be meaningful, the analysis must be repeated from hundreds to thousands of images from several biological replicates, a daunting task. Here we report a custom computer program to analyze key structural features of synapses: SynapsEM. In short, we developed ImageJ/Fiji macros to record x,y-coordinates of segmented structures. The coordinates are then exported as text files. Independent investigators can reload the images and text files to reexamine the segmentation using ImageJ. The Matlab program then calculates and reports key synaptic parameters from the coordinates. Since the values are calculated from coordinates, rather than measured from each micrograph, other parameters such as locations of docked vesicles relative to the center of an active zone can be extracted in Matlab by additional scripting. Thus, this program can accelerate the morphometry of synapses and promote a more comprehensive analysis of synaptic ultrastructure.

A High-Density Human Mitochondrial Proximity Interaction Network

We used BioID, a proximity-dependent biotinylation assay, to interrogate 100 mitochondrial baits from all mitochondrial sub-compartments to create a high resolution human mitochondrial proximity interaction network. We identified 1465 proteins, producing 15626 unique high confidence proximity interactions. Of these, 528 proteins were previously annotated as mitochondrial, nearly half of the mitochondrial proteome defined by Mitocarta 2.0. Bait-bait analysis showed a clear separation of mitochondrial compartments, and correlation analysis among preys across all baits allowed us to identify functional clusters involved in diverse mitochondrial functions, and to assign uncharacterized proteins to specific modules. We demonstrate that this analysis can assign isoforms of the same mitochondrial protein to different mitochondrial sub-compartments, and show that some proteins may have multiple cellular locations. Outer membrane baits showed specific proximity interactions with cytosolic proteins and proteins in other organellar membranes, suggesting specialization of proteins responsible for contact site formation between mitochondria and individual organelles. This proximity network will be a valuable resource for exploring the biology of uncharacterized mitochondrial proteins, the interactions of mitochondria with other cellular organelles, and will provide a framework to interpret alterations in sub-mitochondrial environments associated with mitochondrial disease.

ACTIVITY AND CONTENT OF SUPEROXIDEDISMUTASE IZOZYMES IN GRANULOSE CELLS FROM COW OVARY FOLLICLES

The activity and content of superoxide dismutase isoforms (SOD) in granulosa cells from cow ovarian follicles were studied for research after slaughter of cows ovaries were selected, which according to the physiological state were divided into groups: with "fresh" ovulation, at the site of the ovulated follicle there is a hole, no corpus luteum or diameter up to 5 mm, red color (n = 14); with "early corpus luteum", diameter 10-20 mm, color red or brown (n = 41); with “late corpus luteum", diameter 5–15 mm, color yellow (n = 32); "follicular growth", without the corpus luteum (n = 84). The ovaries of cows with small (&lt;4 mm), medium (4 - 7 mm) and large (&gt; 7 mm) follicles were used. Antral fluid was obtained from the follicles, from which granulosa cells were isolated. Cells were suspended according to the volume of follicular fluid in the medium Dulbeccos modified Eagle medium (DME) with the addition of estrus serum of cows, follicular fluid, insulin and heparin. In cell culture, protein concentration, activity, and superoxide dismutase isozymes were determined. It was found that granulosa cells are characterized by SOD activity - 12.4 ± 0.74 IU / mg protein (6.8 ± 1.72 - 19.8 ± 3.75 IU / mg protein). The activity of SOD in the culture of granulosa cells had 5–6 isoforms of the enzyme. It was found that isoforms at the site of localization are divided into cytosolic, mitochondrial and extracellular proteins of SOD. The cytosolic isoform were represented by 3 - 4, and mitochondrial and extracellular have one active protein of the enzyme. he activity of the enzyme and the content of isoforms depended on the size of the follicles from which the cells are removed and the physiological state of the ovaries. The studied indicators characterize the intensity of oxidative metabolism as a whole in cells and in their individual parts and organelles. For cultivation, it is advisable to select granulosa cells from ovarian follicles of "follicular growth" and "late corpus luteum" because they are characterized by consistently high activity of SOD, which protects intracellular components from the cytotoxic action of superoxide anion.

Spectroscopic and In Vitro Investigations of Boron(III) Complex with Meso-4-Methoxycarbonylpropylsubstituted Dipyrromethene for Fluorescence Bioimaging Applications.

This study focuses on the behavior of a new fluorescent marker for labeling individual biomolecules and staining cell organelles developed on a meso-substituted BODIPY platform. Boron(III) complex with meso-4-methoxycarbonylpropylsubstituted 3,3’,5,5’-tetramethyl-2,2′-dipyrromethene has been synthesized and identified via visible, UV-, NMR- and MS-spectra X-ray. The behavior of fluorophore in solutions has been studied with various experimental techniques. It has been found that luminophore exhibits a high quantum yield (almost ~100–75%) in the blue-green region (513–520 nm) and has high photostability. In addition, biological analysis indicates that the fluorophore exhibits a tendency to effectively penetrate into cell membranes. On the other hand, the proposed BODIPY can be used to study the significant differences among a large number of pathogens of mycotic infections, as well as to visualize structural changes in the plasma membrane, which is necessary for the clearance of mammalian cells undergoing apoptotic cell death.