Zerebralen Aneurysmen sind krankhafte Erweiterung oder Ballonfahren einer lokalisierten Segment eines intrakraniellen Blutgefäßes. Diese Läsionen treten typischerweise bei Verzweigungspunkte der Muttergesellschaft Schiff als Folge der fortschreitenden Schwächung der Arterienwand. Zerebralen Aneurysmen werden meist nach dem Aneurysma platzt verursacht Subarachnoidalblutung nachgewiesen. Diese Form der intrakraniellen Blutung führt ein 45% iges Risiko der Sterblichkeit innerhalb des ersten Monats nach dem Bruch und Überlebenden erleben oft schweren bleibenden neurologischen Verletzungen 1. In jüngster Zeit aufgrund der verstärkten Nutzung moderner bildgebender Verfahren wie MRT und CT sind zerebrale Aneurysmen häufig vor Bruch und Optionen für die Behandlung von zerebralen Aneurysmen sind operatives Clipping-und Coil-Embolisation identifiziert. In diesem Video-Bericht beschreiben wir die mikrochirurgische Clipping eines mittleren Hirnarterie Aneurysma Verwendung eines Ultraschall-perivaskuläre Sonde für Blutgefäße fließen Beurteilung. Das Ziel der chirurgischen Clipping eines zerebralen Aneurysmen ist endgültig entfernen des Aneurysmas aus dem Kreislauf unter Beibehaltung der Blutfluss durch die Muttergesellschaft und distalen Arterien. Typischerweise wird das Operationsmikroskop eingesetzt, um eine verbesserte Darstellung des Aneurysmas bieten und ermöglichen erfolgreiche Präparation des Aneurysmas und Platzierung einer Titan-Clip an der Basis. Unsachgemäße Clip Platzierung kann in Rest-Aneurysma oder teilweisen oder vollständigen Verschluss der distalen Arterien führen, was möglicherweise zu Hirninfarkt. Mehrere Studien mit intraoperative Angiographie während Aneurysmachirurgie haben die Unzulänglichkeit der direkte visuelle Inspektion bei der Vorhersage der Aufrechterhaltung eines adäquaten Blutfluss in der Muttergesellschaft und der distalen Arterien 2,3 demonstriert. Trotz seiner Nützlichkeit, trägt intraoperative Angiographie das Risiko einer arteriellen Verletzung oder Schlaganfall und seine flächendeckende Verfügbarkeit ist begrenzt, 4. In diesem Bericht beschreiben wir die Verwendung eines Ultraschall-Durchfluss-Sonde auf quantitative intraoperative Beurteilung des Blutflusses nach dem chirurgischen Clipping eines Aneurysmas geben. Der Transonic Strömungssonde verwendet Laufzeit-Technologie, um den Blutfluss zu messen. Die Sonde arbeitet mit zwei Sensoren, alternativ Senden und Empfangen von Ultraschall-Signal sowohl im Upstream-und Downstream Richtungen in Bezug auf den Blutfluss 5. Der Durchflussmesser misst dann die Laufzeit für eine Welle von Ultraschall, um den festen Abstand zwischen den Wandlern zu reisen. Der Unterschied zwischen den vor-und nachgelagerten integriert Laufzeiten ist ein Maß für den Blutfluss. Quantitative Durchflussmessungen sind in der übergeordneten und distale Gefäße vor Aneurysma Clipping durchgeführt und dann neu, nachdem Sie den Clip angewendet wurde, um die Erhaltung des Blutflusses 6-8 zu gewährleisten. Wenn die Durchblutung verringert ist, kann den Clip dann neu eingestellt werden wie nötig. Wie in diesem Video, chirurgische Auslöschung einer mittleren Hirnarterie Aneurysma wird durchgeführt und quantitative Beurteilung des Blutflusses mit der Ultraschallsonde ermöglicht die Bestätigung der erhaltenen Durchströmung des distalen hirnversorgenden Arterien, so dass die Vermeidung von neurologischen Komplikationen durch unsachgemäße Wundklammer Platzierung gesehen .

<ol>
	<li>Johnston, S. C., Selvin, S., Gress, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+burden,+trends,+and+demographics+of+mortality+from+subarachnoid+hemorrhage.">The burden, trends, and demographics of mortality from subarachnoid hemorrhage.</a> Neurology. 50, 1413-1418 (1998).</li><li>Alexander, T. D., Macdonald, R. L., Weir, B., Kowalczuk, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraoperative+angiography+in+cerebral+aneurysm+surgery:+a+prospective+study+of+100+craniotomies.">Intraoperative angiography in cerebral aneurysm surgery: a prospective study of 100 craniotomies.</a> Neurosurgery. 39, 10-17 (1996).</li><li>Tang, G., Cawley, C. M., Dion, J. E., Barrow, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraoperative+angiography+during+aneurysm+surgery:+a+prospective+evaluation+of+efficacy.">Intraoperative angiography during aneurysm surgery: a prospective evaluation of efficacy.</a> Journal of neurosurgery. 96, 993-999 (2002).</li><li>Katz, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Is+routine+intraoperative+angiography+in+the+surgical+treatment+of+cerebral+aneurysms+justified?+A+consecutive+series+of+147+aneurysms.">Is routine intraoperative angiography in the surgical treatment of cerebral aneurysms justified? A consecutive series of 147 aneurysms.</a> Neurosurgery. 58, 719-727 (2006).</li><li>Charbel, F. T., Hoffman, W. E., Misra, M., Ostergren, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasonic+perivascular+flow+probe:+technique+and+application+in+neurosurgery.">Ultrasonic perivascular flow probe: technique and application in neurosurgery.</a> Neurological research. 20, 439-442 (1998).</li><li>Fagundes-Pereyra, W. J., Hoffman, W. E., Misra, M., Charbel, F. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clip+readjustment+in+aneurysm+surgery+after+flow+evaluation+using+the+ultrasonic+perivascular+probe:+case+report.+Arquivos+de+neuro-psiquiatria+63.">Clip readjustment in aneurysm surgery after flow evaluation using the ultrasonic perivascular probe: case report. Arquivos de neuro-psiquiatria 63.</a> , 339-344 (2005).</li><li>Amin-Hanjani, S., Meglio, G., Gatto, R., Bauer, A., Charbel, F. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+utility+of+intraoperative+blood+flow+measurement+during+aneurysm+surgery+using+an+ultrasonic+perivascular+flow+probe.">The utility of intraoperative blood flow measurement during aneurysm surgery using an ultrasonic perivascular flow probe.</a> Neurosurgery. 62, 1346-1353 (2008).</li><li>Amin-Hanjani, S., Charbel, F. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flow-assisted+surgical+technique+in+cerebrovascular+surgery.">Flow-assisted surgical technique in cerebrovascular surgery.</a> Surgical neurology. 68, S4-11 (2007).</li></ol>

microsurgical clip obliteration of middle cerebral aneurysm using intraoperative flow assessment

Zerebralen Aneurysmen sind krankhafte Erweiterung oder Ballonfahren einer lokalisierten Segment eines intrakraniellen Blutgefäßes. Operatives Clipping ist eine wichtige Behandlungsoption für Aneurysmen, die den Blutfluss in den aneurysmatischen Segment des Schiffes schließt unter Beibehaltung des Blutflusses in einer normalen Weise versucht. Unsachgemäße Clip Platzierung kann in Rest-Aneurysma mit dem Potenzial für eine spätere Aneurysmaruptur oder teilweisen oder vollständigen Verschluss der distalen Arterien führt Hirninfarkt führen. Hier beschreiben wir die Verwendung eines Ultraschall-Durchfluss-Sonde zur quantitativen Bewertung der arteriellen Blutfluss vor und nach der mikrochirurgischen Clip Platzierung an der Basis eines mittleren Hirnarterie Aneurysma bieten. Diese Information hilft eine ausreichende Aneurysma Rekonstruktion mit der Erhaltung der normalen distalen Durchblutung.

Watch this Scientific Journal Video about Microsurgical Clip Obliteration of Middle Cerebral Aneurysm Using Intraoperative Flow Assessment at JoVE.com

Microsurgical Clip Obliteration of Middle Cerebral Aneurysm Using Intraoperative Flow Assessment

Beschreibung der chirurgischen Auslöschung eines Aneurysma Verwendung eines Ultraschall-Durchfluss-Sonde an den arteriellen Blutfluss vor und nach der Aneurysmen-Clip Platzierung zu bewerten.

Mikrochirurgische Clip Obliteration des Nahen zerebralen Aneurysmen mit Intraoperative Flow-Bewertung

Cerebral aneurysms are abnormal widening or ballooning of a localized segment of an intracranial blood vessel.  Surgical clipping is an important ...

microsurgical-clip-obliteration-middle-cerebral-aneurysm-using

Research

JoVE Journal

Biology

13.6K Aufrufe.  Havard Medical School, Massachusetts General Hospital. Beschreibung der chirurgischen Auslöschung eines Aneurysma Verwendung eines Ultraschall-Durchfluss-Sonde an den arteriellen Blutfluss vor und nach der Aneurysmen-Clip Platzierung zu bewerten.

Serie: Microsurgical Clip Obliteration of Middle Cerebral Aneurysm Using Intraoperative Flow Assessment

Psychophysiological Stress Assessment Using Biofeedback 

In the last half century, research in biofeedback has shown the extent to which the human mind can influence the functioning of the autonomic nervous system, previously thought to be outside of conscious control. By letting people observe signals from their own bodies, biofeedback enables them to develop greater awareness of their physiological and psychological reactions, such as stress, and to learn to modify these reactions. Biofeedback practitioners can facilitate this process by assessing people s reactions to mildly stressful events and formulating a biofeedback-based treatment plan. During stress assessment the practitioner first records a baseline for physiological readings, and then presents the client with several mild stressors, such as a cognitive, physical and emotional stressor. Variety of stressors is presented in order to determine a person's stimulus-response specificity, or differences in each person's reaction to qualitatively different stimuli. This video will demonstrate the process of psychophysiological stress assessment using biofeedback and present general guidelines for treatment planning.

Psychophysiological Stress Assessment Using Biofeedback

In this video we will describe one method of assessing person's psychophysiological reaction to stress using biofeedback. We will also present general guidelines for treatment planning.

Psychophysiologische Stress Assessment mit Biofeedback

In the last half century, research in biofeedback has shown the extent to which the human mind can influence the functioning of the autonomic nervous ...

Forschung

Biologie

A Quantitative Assessment of The Yeast Lipidome using Electrospray Ionization Mass Spectrometry

Lipids are one of the major classes of biomolecules and play important roles membrane dynamics, energy storage, and signalling1-4. The budding yeast Saccharomyces cerevisiae, a genetically and biochemically manipulable unicellular eukaryote with annotated genome and very simple lipidome, is a valuable model for studying biological functions of various lipid species in multicellular eukaryotes2,3,5. S. cerevisiae has 10 major classes of lipids with chain lengths mainly of 16 or 18 carbon atoms and either zero or one degree of unsaturation6,7. Existing methods for lipid identification and quantification - such as high performance liquid chromatography, thin-layer chromatography, fluorescence microscopy, and gas chromatography followed by MS - are well established but have low sensitivity, insufficiently separate various molecular forms of lipids, require lipid derivitization prior to analysis, or can be quite time consuming. Here we present a detailed description of our experimental approach to solve these inherent limitations by using survey-scan ESI/MS for the identification and quantification of the entire complement of lipids in yeast cells. The described method does not require chromatographic separation of complex lipid mixtures recovered from yeast cells, thereby greatly accelerating the process of data acquisition. This method enables lipid identification and quantification at the concentrations as low as g/ml and has been successfully applied to assessing lipidomes of whole yeast cells and their purified organelles. Lipids extraction from whole yeast cells for using this method of lipid analysis takes two to three hours. It takes only five to ten minutes to run each sample of extracted and dried lipids on a Q-TOF mass spectrometer equipped with a nano-electrospray source.

We describe a new quantitative lipidomics method for identifying numerous lipid species in yeast using survey-scan electrospray ionization mass spectrometry (ESI/MS). This method exceeds currently available methods for lipid identification and quantification in the ability to resolve various molecular forms of lipids, sensitivity, and speed.

Eine quantitative Bewertung der Hefe Lipidome mit Elektrospray-Ionisation-Massenspektrometrie

Lipids are one of the major classes of biomolecules and play important roles membrane dynamics, energy storage, and signalling1-4. The budding yeast ...

Platelet Adhesion and Aggregation Under Flow using Microfluidic Flow Cells

Platelet aggregation occurs in response to vascular injury where the extracellular matrix below the endothelium has been exposed. The platelet adhesion cascade takes place in the presence of shear flow, a factor not accounted for in conventional (static) well-plate assays. This article reports on a platelet-aggregation assay utilizing a microfluidic well-plate format to emulate physiological shear flow conditions. Extracellular proteins, collagen I or von Willebrand factor are deposited within the microfluidic channel using active perfusion with a pneumatic pump. The matrix proteins are then washed with buffer and blocked to prepare the microfluidic channel for platelet interactions. Whole blood labeled with fluorescent dye is perfused through the channel at various flow rates in order to achieve platelet activation and aggregation. Inhibitors of platelet aggregation can be added prior to the flow cell experiment to generate IC50 dose response data.

The platelet adhesion cascade takes place in the presence of shear flow, a factor not accounted for in conventional (static) well-plate assays. This article reports on a platelet-aggregation assay utilizing a microfluidic well-plate format to emulate physiological shear flow conditions.

Adhäsion und Aggregation unter Fluss mit Mikrofluidik-Flow-Cells

Platelet aggregation occurs in response to vascular injury where the extracellular matrix below the endothelium has been exposed. The platelet adhesion ...

Surgical Induction of Endolymphatic Hydrops by Obliteration of the Endolymphatic Duct

Surgical induction of endolymphatic hydrops (ELH) in the guinea pig by obliteration and obstruction of the endolymphatic duct is a well-accepted animal model of the condition and an important correlate for human Meniere's disease. In 1965, Robert Kimura and Harold Schuknecht first described an intradural approach for obstruction of the endolymphatic duct (Kimura 1965). Although effective, this technique, which requires penetration of the brain's protective covering, incurred an undesirable level of morbidity and mortality in the animal subjects. Consequently, Andrews and Bohmer developed an extradural approach, which predictably produces fewer of the complications associated with central nervous system (CNS) penetration.(Andrews and Bohmer 1989) 
The extradural approach described here first requires a midline incision in the region of the occiput to expose the underlying muscular layer. We operate only on the right side. After appropriate retraction of the overlying tissue, a horizontal incision is made into the musculature of the right occiput to expose the right temporo-occipital suture line. The bone immediately inferio-lateral the suture line (Fig 1) is then drilled with an otologic drill until the sigmoid sinus becomes visible. Medial retraction of the sigmoid sinus reveals the operculum of the endolymphatic duct, which houses the endolymphatic sac. Drilling medial to the operculum into the area of the endolymphatic sac reveals the endolymphatic duct, which is then packed with bone wax to produce obstruction and ultimately ELH. 
In the following weeks, the animal will demonstrate the progressive, fluctuating hearing loss and histologic evidence of ELH.

This video shows how to surgically obstruct the guinea pig's endolymphatic duct to produce endolymphatic hydrops.

Chirurgische Induktion von Endolymphatischer Hydrops durch Obliteration des Ductus endolymphaticus

Surgical induction of endolymphatic hydrops (ELH) in the guinea pig by obliteration and obstruction of the endolymphatic duct is a well-accepted animal ...

Flow Cytometry Purification of Mouse Meiotic Cells

The heterogeneous nature of cell types in the testis and the absence of meiotic cell culture models have been significant hurdles to the study of the unique differentiation programs that are manifest during meiosis. Two principal methods have been developed to purify, to varying degrees, various meiotic fractions from both adult and immature animals: elutriation or Staput (sedimentation) using BSA and/or percoll gradients. Both of these methods rely on cell size and density to separate meiotic cells1-5. Overall, except for few cell populations6, these protocols fail to yield sufficient purity of the numerous meiotic cell populations that are necessary for detailed molecular analyses. Moreover, with such methods usually one type of meiotic cells can be purified at a given time, which adds an extra level of complexity regarding the reproducibility and homogeneity when comparing meiotic cell samples.
Here, we describe a refined method that allows one to easily visualize, identify, and purify meiotic cells, from germ cells to round spermatids, using FACS combined with Hoechst 33342 staining7,8. This method provides an overall snapshot of the entire meiotic process and allows one to highly purify viable cells from most stage of meiosis. These purified cells can then be analyzed in detail for molecular changes that accompany progression through meiosis, for example changes in gene expression9,10and the dynamics of nucleosome occupancy at hotspots of meiotic recombination11.

An efficient method to obtain highly purified viable meiotic fractions from mouse testis is described, which combines a refined cell dissociation protocol with fluorescent activated cell sorting (FACS). This method takes advantage of differences in the DNA content and nuclear density of discrete meiotic fractions.

Durchflusszytometrie Reinigung der Maus meiotischen Zellen

The heterogeneous nature of cell types in the testis and the absence of meiotic cell culture models have been significant hurdles to the study of the ...

Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes

Fluorescent cell tracking dyes, in combination with flow and image cytometry, are powerful tools with which to study the interactions and fates of different cell types in vitro and in vivo.1-5 Although there are literally thousands of publications using such dyes, some of the most commonly encountered cell tracking applications include monitoring of:
<ol>
	<li>stem and progenitor cell quiescence, proliferation and/or differentiation6-8</li>
	<li>antigen-driven membrane transfer9 and/or precursor cell proliferation3,4,10-18 and</li>
	<li>immune regulatory and effector cell function1,18-21.</li>
</ol>
Commercially available cell tracking dyes vary widely in their chemistries and fluorescence properties but the great majority fall into one of two classes based on their mechanism of cell labeling. "Membrane dyes", typified by PKH26, are highly lipophilic dyes that partition stably but non-covalently into cell membranes1,2,11. "Protein dyes", typified by CFSE, are amino-reactive dyes that form stable covalent bonds with cell proteins4,16,18. Each class has its own advantages and limitations. The key to their successful use, particularly in multicolor studies where multiple dyes are used to track different cell types, is therefore to understand the critical issues enabling optimal use of each class2-4,16,18,24.
The protocols included here highlight three common causes of poor or variable results when using cell-tracking dyes. These are:
<ol>
	<li>Failure to achieve bright, uniform, reproducible labeling. This is a necessary starting point for any cell tracking study but requires attention to different variables when using membrane dyes than when using protein dyes or equilibrium binding reagents such as antibodies.</li>
	<li>Suboptimal fluorochrome combinations and/or failure to include critical compensation controls. Tracking dye fluorescence is typically 102 - 103 times brighter than antibody fluorescence. It is therefore essential to verify that the presence of tracking dye does not compromise the ability to detect other probes being used.</li>
	<li>Failure to obtain a good fit with peak modeling software. Such software allows quantitative comparison of proliferative responses across different populations or stimuli based on precursor frequency or other metrics. Obtaining a good fit, however, requires exclusion of dead/dying cells that can distort dye dilution profiles and matching of the assumptions underlying the model with characteristics of the observed dye dilution profile.</li>
</ol>
Examples given here illustrate how these variables can affect results when using membrane and/or protein dyes to monitor cell proliferation.

Successful use of cell tracking dyes to monitor immune cell function and proliferation involves several critical steps. We describe methods for: 1) obtaining bright, uniform, reproducible label-ing with membrane dyes; 2) selecting fluorochromes and data acquisition conditions; and 3) choosing a model to quantify cell proliferation based on dye dilution.

Optimiert Färbung und Proliferation Modeling-Methoden für die Zellteilung Überwachung mit Cell-Tracking Dyes

Fluorescent cell tracking dyes, in combination with flow and image cytometry, are powerful tools with which to study the interactions and fates of ...

Techniques for the Analysis of Extracellular Vesicles Using Flow Cytometry

Extracellular Vesicles (EVs) are small, membrane-derived vesicles found in bodily fluids that are highly involved in cell-cell communication and help regulate a diverse range of biological processes. Analysis of EVs using flow cytometry (FCM) has been notoriously difficult due to their small size and lack of discrete populations positive for markers of interest. Methods for EV analysis, while considerably improved over the last decade, are still a work in progress. Unfortunately, there is no one-size-fits-all protocol, and several aspects must be considered when determining the most appropriate method to use. Presented here are several different techniques for processing EVs and two protocols for analyzing EVs using either individual detection or a bead-based approach. The methods described here will assist with eliminating the antibody aggregates commonly found in commercial preparations, increasing signal&#8211;to-noise ratio, and setting gates in a rational fashion that minimizes detection of background fluorescence. The first protocol uses an individual detection method that is especially well suited for analyzing a high volume of clinical samples, while the second protocol uses a bead-based approach to capture and detect smaller EVs and exosomes.

Many different methods exist for the measurement of extracellular vesicles (EVs) using flow cytometry (FCM). Several aspects should be considered when determining the most appropriate method to use. Two protocols for measuring EVs are presented, using either individual detection or a bead-based approach.

Techniken für die Analyse der extrazellulären Vesikeln mittels Durchflusszytometrie

Extracellular Vesicles (EVs) are small, membrane-derived vesicles found in bodily fluids that are highly involved in cell-cell communication and help ...

A Protocol for Using Gene Set Enrichment Analysis to Identify the Appropriate Animal Model for Translational Research

Recent studies that compared transcriptomic datasets of human diseases with datasets from mouse models using traditional gene-to-gene comparison techniques resulted in contradictory conclusions regarding the relevance of animal models for translational research. A major reason for the discrepancies between different gene expression analyses is the arbitrary filtering of differentially expressed genes. Furthermore, the comparison of single genes between different species and platforms often is limited by technical variance, leading to misinterpretation of the con/discordance between data from human and animal models. Thus, standardized approaches for systematic data analysis are needed. To overcome subjective gene filtering and ineffective gene-to-gene comparisons, we recently demonstrated that gene set enrichment analysis (GSEA) has the potential to avoid these problems. Therefore, we developed a standardized protocol for the use of GSEA to distinguish between appropriate and inappropriate animal models for translational research. This protocol is not suitable to predict how to design new model systems a-priori, as it requires existing experimental omics data. However, the protocol describes how to interpret existing data in a standardized manner in order to select the most suitable animal model, thus avoiding unnecessary animal experiments and misleading translational studies.

We provide a standardized protocol for the use of gene set enrichment analysis of transcriptomic data to identify an ideal mouse model for translational research. 
This protocol can be used with DNA microarray and RNA sequencing data and can further be extended to other omics data if data are available.

Ein Protokoll für die Verwendung von Gen Set Anreicherung Analyse zur Ermittlung von geeigneten Tiermodell für die translationale Forschung

Recent studies that compared transcriptomic datasets of human diseases with datasets from mouse models using traditional gene-to-gene comparison ...

Fish Sperm Assessment Using Software and Cooling Devices

For gamete quality evaluation, there are innovative, rapid, and quantitative techniques that can provide useful data for aquaculture. Computerized systems for sperm analysis were developed to measure several parameters and one of the most commonly measured is the sperm motility.
Initially, this computer technology was designed for mammalian species, although it can also be used for fish sperm analysis. Fish have specific features that can affect sperm assessment such as a short motility time after activation and, in some cases, adaptation to lower temperatures. Thus, it is necessary to modify both software and hardware components to make motility analysis more efficient for fish sperm analysis. For mammalian sperm, the heating plate is used to maintain optimal temperatures of spermatozoa. However, for some fish species, it is advantageous to use a lower temperature to prolong the duration of motility, since the sperm remain active for less than 2 min. Therefore, cooling devices are necessary to refrigerate samples at constant temperature over the time of analysis, including on the optical microscope. This protocol describes the analysis of fish sperm motility using software for sperm analysis and new cooling devices to optimize the results.

The present protocol describes a procedure of fish sperm assessment using computer-assisted sperm analysis and cooling devices. The software gives a rapid, accurate and quantitative analysis of fish sperm quality based on spermatozoa motility, which can be a useful tool in aquaculture to improve reproduction success.

Fisch Sperma Bewertung mit Software und Kühlgeräte

For gamete quality evaluation, there are innovative, rapid, and quantitative techniques that can provide useful data for aquaculture. Computerized systems ...

Microsurgical Obstruction of Testes Fusion in Spodoptera litura

Instead of using genetic methods like RNA interference (RNAi) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated endonuclease Cas9, a physical barrier was microsurgically inserted between the testes of Spodoptera litura to study the impact of this microsurgery on its growth and reproduction. After inserting aluminum foil between the testes, insect molting during metamorphosis proceeded normally. Insect growth and development were not remarkably altered; however, the number of sperm bundles changed if testes fusion was stopped by the microsurgery. These findings imply that blocking testicular fusion can influence male reproduction capability. The method can be further applied to interrupt communication between organs to study the function of specific signaling pathways. Compared to conventional surgery, microsurgery only requires freezing anesthetization, which is preferable to carbon dioxide anesthetization. Microsurgery also minimizes the surgery site area and facilitates wound healing. However, the selection of materials with specific functions needs further investigation. Avoiding tissue injury is crucial when making incisions during the operation.

Microsurgical Obstruction of Testes Fusion in Spodoptera litura

Aluminum foil was microsurgically inserted between the testes of Spodoptera litura to obstruct the fusion of testis. The procedure includes freezing, fixing, disinfection, incision, placing the barrier, suturing, postoperative feeding, and inspection. This approach provides a method to interfere with tissue formation.

Mikrochirurgische Obstruktion der Hodenfusion bei Spodoptera litura

Instead of using genetic methods like RNA interference (RNAi) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated ...