In contrast, GWF using SpermFertil® columns significantly reduced the number of EPS+ spermatozoa (−8.8% EPS + sperm, P < 0.01) with control without alteration of the MMP and without activation of caspase 3 (P > 0.05). After cryopreservation, motility (WHOa+b) decreased to 36.1 ± 3.1% (P < 0.001). Products by Application. Methods of sperm preparation for assisted reproduction. Semen samples prepared by simple wash, standard GWF and annexin V-MACS served as controls. MDL number MFCD00147032. 020 487 70 70 - benelux.info@thermofisher.com It can be a loose fill material, blown into attics, or, together with an active binder sprayed on … 1996 Sep;66(3):459-62. doi: 10.1016/s0015-0282(16)58519-7. Annexin V GWF is as easy to perform as conventional GWF and requires significantly less time and processing steps, compared to the MACS procedure. All fluorescence signals of the labelled spermatozoa were analysed by the FACScan (Becton Dickinson, San Jose, CA, USA). MACS separation of sperm yields two fractions: annexin V-negative (intact membranes, spermatozoa free of apoptotic markers) and annexin V-positive (EPS, spermatozoa-labelled positive for apoptotic markers) which is retained in the magnetic field (Grunewald et al., 2001; Paasch et al., 2003b). The cervical mucus selectively allows only progressively motile sperm of normal shape and size to penetrate and migrate through the cervix. The oocytes from each patient were randomly injected, half with spermatozoa prepared by glass wool filtration and half with Percoll-separated spermatozoa from the same semen sample. Descriptive statistics of apoptotic markers detected in all sperm fractions. ... (1980) Glass Wool Filtration of Human Semen. Filter Wool helps keep water clear and free of impurities, by replacing weekly it allows you to easily remove any foreign objects, like left over food, plant leaves from your tank to reduce the load on your bacterial filtration. The procedure allows the enrichment of a sperm population free of apoptosis markers, giving higher fertilization potential. Therefore, raw glass wool may not be used for sperm preparation prior to ART (Henkel and Schill, 2003). Out of 97 metaphase II oocytes injected with Percoll-selected spermatozoa, 71 fertilized and 69 cleaved. The excitation wavelength was 488 nm, supplied by an argon laser at 15 mW. Although our data support the highly efficient filtration capacity of a solid phase annexin V coated glass wool filter, further development is needed for optimization. Minimal EPS occurred in the annexin V-negative fraction, whereas a sizeable number of spermatozoa that stained negative for PS were found in the annexin V-positive fraction. Epub 2011 Dec 5. Urology-andrology Glass wool column filtration versus mini-Percoll gradient for processing poor quality semen samples* * Presented at the 50th Annual Meeting of The American Fertility Society, San Antonio, Texas, November 5 to 10, 1994. Semen samples (n = 42) were subjected to a molecular glass wool filter system using glass surfaces coated with annexin V and compared with aliquots separated by conventional glass wool, as well as with annexin V-MACS.The extent of apoptosis was assessed by measuring levels of activated caspase 3 using fluorescein-labelled inhibitors of caspase, alterations in mitochondrial membrane … Finally, the tubes were plunged into a liquid nitrogen tank (Biosafe MD, Cryotherm Kirchen, Germany) and stored at −196°C in the gas phase. Also, the percentage of normal forms was 1.25-3.31% higher after glass wool filtration but was poorly correlated to the method (r = 0.47, P = 0.017). Sonja Grunewald, Werner Miska, Gisela Miska, Manja Rasch, Martin Reinhardt, Hans-Juergen Glander, Uwe Paasch, Molecular glass wool filtration as a new tool for sperm preparation, Human Reproduction, Volume 22, Issue 5, May 2007, Pages 1405–1412, https://doi.org/10.1093/humrep/dem015. The inhibitor was used with the appropriate controls according to the kit instructions provided by the manufacturer (Carboxyfluorescein FLICA, Immunochemistry Technologies, Bloomington, MN, USA). Spray Booth Filter, Paint Booth Floor Filter, Filter Fiberglass Floor manufacturer / supplier in China, offering Glass Wool Filter for Paint Extraction Spray Booth, Operating Room HEPA Filter, Pm2.5 High Efficiency H10 H11 H12 H13 Activated Carbon Honeycomb Composite Replaceable Air HEPA Filter and … After the supernatant was discarded, each sperm pellet was incubated with 50 µl of secondary antibody (goat anti-mouse immunoglobulin G, fluorescein conjugate, Upstate Cell Signaling Solutions, Lake Placid, NY, USA) on ice for 20 min and protected from direct light. Products by Industry. Successful assisted reproduction in male infertility is mainly dependent on the quality of the sperm sample (Ombelet et al., 2003). Bottle Tops and Filter Units; Extraction Thimbles; Filter Holders and Hardware; Filter Paper; Filtering Funnels; Food and Beverage Testing; ... Glass Wool; Glass Wool 1 – 30 1132 . Spermatozoa were incubated with annexin V-conjugated microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany) for 15 min at room temperature. The disruption of the MMP and the EPS to the sperm surface showed also a significant positive correlation (R = 0.30, P < 0.01). The extent of activated caspase 3, intact MMP and EPS were evaluated by FACS analyses. These results were not statistically different. An example of when glass wool is essential is the filtration of certain ionic liquids which dissolve filter paper! The successful fixation of annexin V to the glass wool was evaluated by the relative presence of annexin V on the glass wool surface of an untreated SpermFertil® column in comparison to an annexin V glass wool column. Vials were thawed by incubating at 37°C for 20 min. This new sperm preparation strategy enables enrichment of vital spermatozoa free of apoptotic markers, especially when cryopreservation needs to be performed (Grunewald et al., 2001; Paasch et al., 2004c). Semen samples (n = 42) were subjected to a molecular glass wool filter system using glass surfaces coated with annexin V and compared with aliquots separated by conventional glass wool, as well as with annexin V-MACS. Asian J Androl. It was concluded that glass wool filtration for semen preparation in an ICSI programme offers higher sperm recovery and sperm morphology of superior quality than with the classic two-layer Percoll gradient method, without affecting the fertilization rate and embryo quality. Similarly, the total number of spermatozoa available for ICSI was 0.18 x 10(6)-2.44 x 10(6) higher with the glass wool column and was highly correlated to the method (r = 0.956, P = 0.0001). Light microscopy and fluorescence microscopy of the annexin V glass wool (A) and the standard SpermFertil® column (B) after incubation with the fluorescein isothiocyanate-labelled anti-annexin V antibody. The fixation of annexin V on the surface of the glass wool was proven by the receipt of strong fluorescence signals from annexin V glass fibers after incubation with the FITC-labelled anti-human annexin V antibody (Figure 2A, red arrows). Get contact details & address of companies manufacturing and supplying Glass Wool, Glass Wool Insulation, Glasswool across India. Half of each of 25 semen samples were prepared by each technique. Therefore, unlike routine sperm preparation techniques, sperm preparation using annexin V coupled super-paramagnetic microbeads acts on the molecular level. Therefore, annexin V binding to spermatozoa may be used to label sperm that have compromised membrane integrity and that are less capable of fertilizing oocytes (Glander and Schaller, 1999). The comparative pregnancy rates for the groups were A1 (7.1%) versus A2 (7.1%) and B (28.9%) versus C (31.6%). In addition, the conventional SpermFertil® glass wool is at least partially able to separate spermatozoa with cryopreservation and thawing induced membrane damage, as well as following the activation of caspases and the disruption of MMP. Annexin V-negative spermatozoa were separated by magnetic-activated cell sorting (MACS) or by annexin V-coated glass wool following one-step wash and re-suspension in human tubal fluid (HTF) media. The extent of apoptosis was assessed by measuring levels of activated caspase 3 using fluorescein-labelled inhibitors of caspase, alterations in mitochondrial membrane potential (MMP) using a lipophilic cationic dye, and EPS using a fluorescein isothiocyanate-coupled monoclonal antibody. Overall experimental design showing the different steps performed in our study. Only when applied to cryopreserved samples was there a minor filtration effect in terms of apoptosis markers. Find here Glass Wool, Glass Wool Insulation manufacturers, suppliers & exporters in India. Following this, the 2 ml of the semen sample was loaded on top of the column. The green light in B is a result of a prolongation of exposure time to >5 s. The application of the same exposure time as in A resulted in a black picture. Green fluorescence (480–530 nm) was measured in the FL-1 channel and red fluorescence (580–630 nm) in the FL-2 channel. Subsequently, both columns were rinsed with 5 ml HTF media. Superscripts 1–6 indicate significantly different when compared with: (1) aliquot 1, (2) aliquot 2, (3) aliquot 3, (4) aliguot 4, (5) aliquot 5 and (6) aliquot 6 (P < 0.05 was considered significant using Wilcoxon-test). A second washing step in PBSB (300 g for 5 min at 20°C) was performed to remove excess antibody that was not bound to the spermatozoal surface. The main apoptotic event detectable at the sperm surface is the externalization of the phospholipid PS, which is usually present only on the inner leaflet of the sperm plasma membrane (Vermes et al., 1995; Oosterhuis and Vermes, 2004). Please enable it to take advantage of the complete set of features! Axis Solutions Private Limited - Offering Glass Wool Filter, NS DUTTA Filters, filter Concept Filters, BAPL Filters, SAP Filters Filters, DP Engineers Filters, Gas Analysis Components in Kathwada, Ahmedabad, Gujarat. Through the sieve pores, only water will pass. activated caspases, externalization of phosphatidylserine (EPS) at the outer plasma membrane, DNA-fragmentation and disruption of the mitochondrial membrane potential (MMP) (Comhaire et al., 1988; Glander and Schaller, 1999; Oehninger et al., 2003; Paasch et al., 2003b; Grunewald et al., 2005). Would you like email updates of new search results? Caspase 3 activation and the disruption of MMP exceeded the levels measured in controls. When the total sperm count exceeded 100 × 106 spermatozoa (n = 14) one extra aliquot was subjected to cryopreservation. Fertil Steril. Among the sperm preparation techniques available for ART, GWF is also known to result in semen samples with superior recovery rates, motility and morphology (Van Den et al., 1997; Henkel and Schill, 2003). glass wool filter is composed of processed silica that is packed into a wool column. Positive controls were performed using betulinic acid (60 µg ml−1) (Paasch et al., 2004b). Saturation of annexin V binding was achieved at 100 µg ml−1. 21 The tip was removed from a 1 ml Becton Dickinson sy­ ringe and 15 mg of glass wool (microfiber … For Permissions, please email: journals.permissions@oxfordjournals.org, Qualitative research methods: when to use them and how to judge them, Bleeding patterns after vaginal misoprostol for treatment of early pregnancy failure, Vaginal misoprostol alone for medical abortion up to 9 weeks of gestation: efficacy and acceptability, Early pregnancy termination with vaginal misoprostol before and after 42 days gestation, Low-level sex chromosome mosaicism in female partners of couples undergoing ICSI therapy does not significantly affect treatment outcome, About the European Society of Human Reproduction and Embryology, Society for Assisted Reproductive Technology and American Society for Reproductive Medicine, 2004, Receive exclusive offers and updates from Oxford Academic, Glass wool column filtration of human semen: relation to swim-up procedure and outcome of IVF, Quantitative study of caspase-3 activity in semen and after swim-up preparation in relation to sperm quality, Fruit and vegetable intake and their pesticide residues in relation to semen quality among men from a fertility clinic. NACRES NA.22 . Dye aggregates cannot be formed in the absence of the membrane potential. Negative controls were processed identically for each fraction except that the antibody was replaced with equal amounts of PBS. Spermatozoa with intact mitochondria (preserved MMP) emit an intense red fluorescence due to the formation of the dye aggregates. Glass wool is produced in rolls or in slabs, with different thermal and mechanical properties. Oxford University Press is a department of the University of Oxford. Although there was no difference between both annexin V-based preparation methods in terms of caspase 3 activation and integrity of MMP (P > 0.05), the annexin V glass wool filtrated subpopulation showed improved reduction of EPS+ spermatozoa, compared with annexin V-MACS (−3.4% EPS+ compared with the MACS system). Owing to the physical characteristics of glass wool, the fluorescence signals were transmitted along the fibers resulting in a homogeneous fluorescence on their surface (Figure 2A). Activated caspase 3 levels, integrity of the MMP and extent of EPS were assessed in the control aliquot (simple HTF wash) and in all aliquots following the different preparation procedures (Figure 1). Moreover, Glass wool is the most common source of insulation in ceilings of residential buildings, and … NLM The power of the magnetic field was measured as 0.5 T between the poles of the magnet and up to 1.5 T within the iron globes of the column. This may be attributed to the fact that binding sites are already blocked by the annexin V microbeads. Alibaba.com offers 832 glass wool filtration products. A reduction of peroxidase-positive cells of up to 90% can be achieved (Sanchez et al., 1996). Sephadex and glass wool filtration samples were filtered by placing columns in a water bath at 37 for 5~10 min. However, the method apparently induced significant caspase 3 activation (+13.2% CP3+ sperm, P < 0.05), as well as elevated degrees of disruption of the MMP (+13.6% sperm with disrupted MMP compared with HTF wash control, P > 0.05). Raw glass wool had a clear negative impact on semen samples. After being washed with 1 ml HTF media, the columns were incubated with 1 mL fluorescein isothiocyanate (FITC)-labelled anti-human annexin V and clone VAC-α diluted 1:20 in PBS pH 7.4 (Bender MedSystems, Vienna, Austria) for 20 min at room temperature. This newly developed solid phase molecular filter system has been proven to enrich spermatozoa free of apoptosis markers to the same extent as the annexin V magnetic separation technique. Hammadeh ME, Kühnen A, Amer AS, Rosenbaum P, Schmidt W. Int J Androl. Key Benefits: Removes free floating debris for a … Reduction of the dithiogroup was achieved by the addition of 500 µl dithiothreitol (DTT) (50 mM in PBS) for 1 h at room temperature. Filtration using raw glass wool significantly reduced the number of EPS+ spermatozoa in comparison to the HTF wash control (−4.9% EPS+ sperm, P < 0.05, Table I). Testing and Filtration. 2001 Dec;24(6):360-8. doi: 10.1046/j.1365-2605.2001.0317a.x. Furthermore, sperm prepared according to this protocol showed an improved ability to fertilize oocytes using the hamster oocyte penetration assay (Said et al., 2005c). Recently, samples processed by GWF using the SpermFertil® system have been demonstrated to enrich sperm with low levels of caspase 3 activation and EPS (Rasch et al., 2005). Therefore, a sperm preparation system using the binding properties of annexin V-without free flotation of super-paramagnetic microbeads in a liquid phase buffer is needed. Epub 2007 Mar 2. After thorough washing with PBS, incubation with 200 µl of SPDP (1 mg ml) in sodium acetate buffer (SAB) (50 mM, pH 4.5) for 1 h at room temperature followed. Annexin V-negative spermatozoa were separated by MACS or by annexin V-coated glass wool following one-step wash and re-suspension in HTF media; n.d., not done.. The sperm population was gated using 90° and forward-angle light scatter to exclude debris and aggregates. Finally, washing with SAB, incubation with annexin V (100 µg ml in PBS) and subsequent washing with PBS plus 1 mM DTT completed the synthesis of annexin V glass wool columns. Applications of glass wool include structural insulation, pipe insulation, filtration and soundproofing. Spermatozoa were incubated with the PS antibody at a final concentration of 0.5 µg ml−1 in PBS containing 2% bovine serum albumin (PBSB) for 20 min on ice, followed by addition of 150 µl PBSB and centrifugation at 300 g for 5 min at 20°C. The annexin V-negative fractions separated by MACS as well as by the newly developed molecular GWF system displayed a lower percentage of caspase 3 activation as well as higher mitochondrial membrane integrity, relative to the classical sperm preparation techniques. Here we report on filter effectiveness in concentrating waterborne human enterovirus, Salmonella enterica, Cryptosporidium parvum, and avian influenza virus. The selection of spermatozoa free of apoptosis markers by molecular glass wool filters may enhance the results of IVF. Epigenome-wide association study (EWAS) for potential transgenerational disease epigenetic biomarkers in sperm following ancestral exposure to the pesticide methoxychlor. NIH A variety of sperm preparation techniques have been proven to select spermatozoa that are characterized by superior motility and morphology and are capable of fertilizing the oocyte. However, annexin V glass wool was as effective as SpermFertil® columns in decreasing the EPS+ sperm when compared with control. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. A glass wool column which increases the motility of the spermatozoa while removing dead and agglutinated spermatozoa and debris from the ejaculate has been devised and the results from its use are encouraging for specific problems. Hypothesis testing was two-tailed, and P values <0.05 were considered statistically significant. For cryopreservation, the semen samples were diluted drop-wise with an equivalent volume of TYB containing 12% (v/v) glycerol. Semen analysis was performed according to the World Health Organization (WHO) guidelines (World Health Organization, 1999). Glass wool aliquots from both columns were prepared on glass slides, covered with a cover slip and evaluated by fluorescence microscopy (Figure 2A and B). Although sperm recovery was similar using glass wool filtration and Percoll gradient in both oligo­ zoospermic and oligoasthenozoospermic samples, glass wool column filtration seemed to recover a higher viable sperm population based on the hypo­ All test aliquots and controls (with 100 µl PBS) were incubated at 37°C for 1 h with 10 µl of the working solution and subsequently washed twice with the rinse buffer. Industries. Samples with ≥50 × 106 spermatozoa-ml−1 and at least 50% progressive sperm motility were selected for the study. Following freezing and thawing of HTF wash samples, the percentage of spermatozoa displaying activated apoptosis signalling increased significantly: + 23.2% sperm containing active caspase 3, + 30.0% sperm with disrupted MMP and + 4.7% EPS+ sperm were detected in the cryopreserved samples (P < 0.01 compared with HTF in Tables I, and II). USA.gov. The increase in ART applications with concurrent low success rates has directed interest to the development of novel approaches of sperm preparation, such as electrophoretic separation and magnetic cell separation, which have demonstrated encouraging results for the isolation of higher quality spermatozoa (Ainsworth et al., 2005). This clearly indicates that the sperm quality can still be improved by integrating annexin V binding as a part of the sperm preparation protocol, which is concordant to our earlier studies (Said et al., 2005a). The kit was used according to the instructions of the manufacturer. Apparently, tightly packed soda lime glass wool filtration yielded a particularly high recovery of motile spermato- Moreover, standard GWF using the SpermFertil® system showed comparable low rates of EPS. Glass wool filtration was initially developed by Paulson and Polakoski [14] in an attempt to decrease the viscosity of semen and to increase sperm motility percentage. The liquid which has obtained after filtration is called the filtrate; in this case, water is the filtrate. Glass wool is a versatile material that can be used for the insulation of walls, roofs and floors. The fixation of annexin V on the surface of the glass wool was demonstrated by strong fluorescence signals (A, red arrows). Our aim was to determine if the annexin V binding principle can be transferred onto a glass wool filter system in order to produce a solid phase filter. Descriptive statistics of apoptotic markers detected in all cryopreserved and thawed sperm fractions. MACS using annexin V microbeads has been repeatedly shown to efficiently isolate non-apoptotic spermatozoa with a superior fertilization capacity (Paasch et al., 2004c; Said et al., 2005c). The percentage of caspase 3 positive (CP3+) spermatozoa was negatively correlated with the percentage of spermatozoa with intact MMP (R = −0.56, P < 0.01, Spearman's rank correlation) and positively correlated with the percentage of spermatozoa with EPS (R = 0.31, P < 0.01). Within the annexin V-positive fraction after MACS, spermatozoa characterized by significantly increased caspase 3 activation, disruption of MMP and EPS were enriched (P < 0.05, compared with sperm washed in HTF, filtrated by raw and SpermFertil® glass wool as well as with the annexin V-negative subpopulations). The technique, transferred to a solid phase GWF system, appears to be adequate and even highly efficient, since the number of spermatozoa with EPS was lowest in the annexin V-negative fraction after molecular GWF. EPS was examined using a monoclonal mouse anti-human PS antibody, clone 1H6 (Upstate Cell Signaling Solutions, Lake Placid, NY, USA). This site needs JavaScript to work properly. The characteristics of the glass wool filtration technique for the preparation of semen samples in an intracytoplasmic sperm injection programme (ICSI) were compared with a two-layer Percoll density gradient procedure.  |  The annexin V-positive fraction after MACS was not investigated, as the enrichment of spermatozoa with an activated apoptosis signalling after cryopreservation and thawing in that fraction has been proven in several previous studies (Grunewald et al., 2001; Paasch et al., 2004c; Grunewald et al., 2005). The fraction composed of apoptotic spermatozoa was retained in the separation column and labelled as annexin V-positive, whereas the fraction with the intact membranes that was eluted through the column was labelled as annexin V-negative. In this study, we present a newly developed solid phase molecular filtration system which combines classical GWF with PS-binding properties of annexin V. The separation effect of the system was evaluated in terms of apoptosis-related parameters. Comparison of sperm preparation methods: effect on chromatin and morphology recovery rates and their consequences on the clinical outcome after in vitro fertilization embryo transfer. A wide variety of glass wool filtration options are available to you, such as warranty of core components, condition, and local service location. In addition, the glass wool filtration (GWF) technique is known to provide sperm samples with comparable recovery rates, motility, morphology and fertilizing capacity if appropriate types of glass wools used (Holmgren and Jeyendran, 1993; Johnson et al., 1996; Van Den et al., 1997; Henkel and Schill, 2003;). In conclusion, annexin V glass wool is easy to use in molecular-based separation method with a high capacity to exclude spermatozoa with activated apoptosis signalling from semen samples. The mean +/- SEM embryo quality score for the glass wool group (2.90 +/- 0.27) was the same as that for the Percoll group (2.80 +/- 0.24). An improved method of sperm selection by glass wool filtration is introduced. As the annexin V-based MACS was proven to enhance sperm quality and oocyte penetration capacity (Said et al., 2005c), application of annexin V glass wool may improve the outcome of ARTS. Annexin V-conjugated microbeads specifically target spermatozoa with deteriorated membranes that display EPS as a manifestation of apoptosis signalling (Paasch et al., 2003c, 2004a,b). 1992 Jul;21(4):517-24. No fertilization failures occurred and 11 patients (44% per oocyte retrieval) became pregnant. The system can be easily integrated into standard semen preparation techniques, providing higher fertilization rates (Said et al., 2005b,c). Briefly, all aliquots were incubated at 37°C for 20 min in 1 µg of the lipophilic cation diluted in 1 ml PBS. To Email: From Email: Message: 18421 Sigma-Aldrich Glass wool for laboratory use CAS Number 65997-17-3. Glass Wool Non-flammable glass wool, suitable for both general laboratory and industrial applications, as well as filtering, is a thermal insulation created from flexible, intertwined borosilicate glass fibers. Available filtration methods to concentrate waterborne viruses are either too costly for studies requiring large numbers of samples, limited to small sample volumes, or not very portable for routine field applications. SpermFertil® GWF columns were purchased from TransMIT GmbH. Read about company. Glass wool for filtration application from Chongqing Yangkai Import & Export Trade Co., Ltd.. Search High Quality Glass wool for filtration application Manufacturing and Exporting supplier on Alibaba.com. The samples were placed into 2.0 ml cryotubes (Faust Laborbedarf AG, Schaffhausen, Switzerland) and frozen with the Nicool LM 10 system (Air Liquide, Wiesbaden, Germany). Chemicals and Labware > Inorganic Reagents > Safety Products and General Applications > Absorption and Filtration > Filtration > Glass Wool : Recently Viewed. PRODUCTS. A minimum of 10 000 spermatozoa were examined for each assay at a flow rate of <100 cells s−1. For assessment by fluorescence-activated cell sorter (FACS), sperm pellets were diluted in 400 µl PBSB. 2009 Sep;10(3):249-55. doi: 10.4142/jvs.2009.10.3.249. The mechanism by which GWF removes leukocytes and deficient spermatozoa with structurally abnormal and physically damaged membranes is thought to be by adherence to the glass wool fibers. Initially, the standard glass wool filtration method, consisting of an adsorption to the positively charged glass wool matrix and subsequent elution with GBE buffer at pH, … Glass wool CAS 65997-17-3 - Find MSDS or SDS, a COA, data sheets and more information. HTF wash, human tubal fluid media wash; raw GW, raw glass wool; SpermFertil® GW, SpermFertil ® glass wool system; annexin V GW, annexin V glass wool system; MACS, magnetic-activated cell sorting using annexin V-coupled super-paramagnetic microbeads; EPS, externalized phosphatidylserine; CP 3, activated caspase 3; MMP, mitochondrial membrane potential. Positive controls were performed by induction of membrane damage with liquid nitrogen.

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