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Shivani Scientific Industries (P) Ltd.




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Shivani Scientific Industries (P) Ltd.

                                         The Spermfuge facilitates user selectable...
Shivani Scientific Industries (P) Ltd.

                                         The motor is extremely smooth even during...
Shivani Scientific Industries (P) Ltd.




                                                                          Autoc...
Shivani Scientific Industries (P) Ltd.




                                          Embryology Academy of research Traini...
Shivani Scientific Industries (P) Ltd.

                                          for the Reproductive Sciences, Australia...
Shivani Scientific Industries (P) Ltd.




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Shivani Scientific Industries (P) Ltd.

                                         mobile nest continues to log for the powe...
Shivani Scientific Industries (P) Ltd.

                                         battery is weak. When the Nest is replace...
Shivani Scientific Industries (P) Ltd.




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Shivani Scientific Industries (P) Ltd.

                                         LAB + GUARD not only protects embryos and...
Shivani Scientific Industries (P) Ltd.




http://www.shivaniscientific.com/ivfproducts/labplusguard.htm[5/13/2009 1:52:02...
Shivani Scientific Industries (P) Ltd.




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Shivani Scientific Industries (P) Ltd.




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Shivani Scientific Industries (P) Ltd.




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Shivani Scientific Industries (P) Ltd.




                                         3D Lab design / 3 D Walk through
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Shivani Scientific Industries (P) Ltd.




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Shivani Scientific Industries (P) Ltd.

                                         C and also when culture media which are u...
Shivani Scientific Industries (P) Ltd.




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Temperature Controlled Centrifugation: -
The Effect on Motion Characteristics of Human Sperm


                      Dr Fr...
Temperature Controlled Centrifugation



                                                                            The f...
Temperature Controlled Centrifugation



Table 3           Comparison of the velocity distribution of sperm cells( %) prep...
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Products and services for reproductive / infertility professionals

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Shivani Scientific Industries is exporter, manufacture and supplier of ivf lab equipments like microinjection tools, ivf workstations,
Spermfuge,ThermoPlate,Embryo Freezer,System for ART,Suction Pumps for oocyte aspiration,CO2 incubator,IVF warming block,IVF

micromanipulator,IVF Microscope,IVF Micropipettes,Clean Air System, Spermfuge,Mobile Nest,Lab Guard,Warming Blocks,
Turnkey Projects,Rapid Recovery System and lab designs to IVF clinics worldwide. We offer products for IVF ART ICSI including

micropipettes,microscopes etc.

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Shivani Scientific Industries Pvt. Ltd.
Telephone:+91 22 28456768 / 2845 7140 / 84 / 5570 2442
Fax:+91 22 28456766 / 28457059
info@shivaniscientific.com
http://www.shivaniscientific.com

we sell our poroducts to reproductive / infertility professionals, consulting and training, laboratory products and services, software, consulting and training, semen analysis, sperm testing, fertility testing, cryopreservation, artificial insemination, donor insemination, cell culture media, microscopy, micromanipulation, embryo transfer and incubation, preimplementation genetic diagnosis, PGD, electronic medical record software, practice management software, blastocyst human serum albumin andrology, immunodiagnostic testing, and IVF culture, Biofertec, Embryotech Laboratories, FertiPro N.V., IMMUCON, Diagnostic Products Corporation, Irvine Scientific, Medical Electronic Systems, Repromedix, Vitrolife Fertility Systems , Sutter Instrument Company, Infertility Database Systems.

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Products and services for reproductive / infertility professionals

  1. 1. Shivani Scientific Industries (P) Ltd. IVF Products » Spermfuge Mobile Nest Lab + Guard Warming Blocks Turnkey Projects Rapid Recovery System IVF Division Click here for Clinical Research paper on SpermFuge Concept Spermfuge is an improvised centrifuge, dedicated totally for elevating the total motile sperm recovery from the parent semen sample, by controlling/maintaining the main parameter of quot;Temperaturequot; thus aimed at Enhancing the ART results. The instrument has been designed to regulate and subsequently maintain the quot;Criticalquot; inner chamber temperature before, after and during centrifugation. Maintenance of this critical temperature throughout the entire sperm processing procedure helps the sperm maintain their peak motility and enhanced linearity. The two of the most vital parameters which act as a yard stick to gauge the effectiveness of an ideal sperm wash. The existing dependency on RPM will be eliminated by on screen g-force selection. Design The Spermfuge has been designed after taking into consideration detailed cellular physiological demands, locomotion kinetics and pooling together ideal suggestions, opinions, ideas and propositions of numerous Andrologists and Embryologists and other professionals in the field of Human Reproduction. The Spermfuge with its Temperature controlled chamber (room temperature to 42°C), programmable microprocessor based controls, multiple tube selection, accurate g-force selection, independent centrifugation and/or heating selection and other added advantages make it an ideal state of the art instrument for use in an ART laboratory. Temperature and Centrifugation The heating range of Spermfuge's chamber is from 2°C above ambient to 42°C. The intelligent control system does not allow the centrifugation to commence till the set temperature is attained. However, one does have the option to commence the centrifugation without chamber heating and to commence the chamber heating without centrifugation. Multiple Programs http://www.shivaniscientific.com/ivfproducts/spermfuge.htm[5/13/2009 1:50:55 PM]
  2. 2. Shivani Scientific Industries (P) Ltd. The Spermfuge facilitates user selectable 9 programs for different applications and also for different set of protocols as standardized by various clinics, laboratories and regulatory bodies. The user can select RCF, RPM, Tube Size, Temperature, Time for each program by a digital encoder. Acceleration and Braking An adjustable brake is provided to prevent the rotor from stopping too quickly at the end of run which might cause disturb to the firmly formed pellet. One can select 3 mode in acceleration and braking, slow medium and to fast can be given on screen. G - Force The exact indication of G - Force helps the ideal sperm pellet formation by subjecting the sperm to an exact and precise amount of G - Force unlike other Centrifuges where G - Force is subject to RPM and the tube size. Now a days centrifugation speeds are never reported in RPM as those numbers are not useful for any one wanting to reproduce the method. (Being dependent Rotor radius.) Rotor Head and Tube Selection Spermfuge has a swing out rotor head to accommodate 8 tubes maximum of 15 ml. The Sperm Fuge can accommodate 6ml Tubes (Falcon 2003), 14 ml Tubes (Falcon 2001) and 15 ml Tubes (Falcon 2095). The tube selection can be done on screen with the help of the encoder, which ensures an exact G - Force selection. Operating Security The Vision port in the lead at the center helps to confirm rotational speed with a stroboscopic device. The safety interlock in the lead prevents accidental opening once the Rotor is operational. The maximum RCF is 800g which prevents accidental damage to the sperm. Intelligent Control System State of the art micro-controller with optomagnetic encoder provides intelligent control system. Digital programming via microprocessor control for various parameters including temperature, RCF and time. The parameters are displayed on four line LCD display. Maintenance The Spermfuge is virtually maintenance free since it utilizes brushless drive motor. http://www.shivaniscientific.com/ivfproducts/spermfuge.htm[5/13/2009 1:50:55 PM]
  3. 3. Shivani Scientific Industries (P) Ltd. The motor is extremely smooth even during running at the maximum speed. The motor is electrically controlled during the acceleration and breaking with smoothness. http://www.shivaniscientific.com/ivfproducts/spermfuge.htm[5/13/2009 1:50:55 PM]
  4. 4. Shivani Scientific Industries (P) Ltd. Autoclaves & Sterilisers » Front Loading Flash Sterliser Model : FL 17 Double Wall Autoclave Laboratory Autoclaves Semi Automatic Portable Autoclave Semi Automatic Gas cum Electric Autoclave Horizontal Sterilizer Fully Automatic Autoclaves GMP Compliant Series Acculeak Detector Features: Table top front loading autoclave Fully Automatic micro-processor based control Sterilization chamber made up of thick stainless steel sheet, tested for high pressure Digital timer for both wet and dry cycle Auto drain of water to reservoir tank & also condensation of steam Alarm after the total command is completed Cycle time 10-15 minutes http://www.shivaniscientific.com/autoclavesteriliser/frontload.htm[5/13/2009 1:51:12 PM]
  5. 5. Shivani Scientific Industries (P) Ltd. Embryology Academy of research Training (EART), a part of Shivani Scientific Inductries, is a Young Emerging New Dimension in the field of ART Education. EART initiated sincere and honest efforts to understand current needs of the embryologists & gynecologists for learning various ART techniques (IUI, IVF, ICSI and Crayopreservation). Thus, EART provides a platform for perfecting Assisted Reproductive skills & Latest technologies. EART conducts various hands on courses in India with the help of renowned Indian and International faculties. EART has successfully ensured that each participant gets an adequate hands-on experience and project mock cycles, coupled with the requisite theory lectures. One of the basic criteria of learning is getting an actual hands-on experience. The hands-on session details are mentioned in the data sheets of each course. One-on-one training offers the maximum opportunity to learn at EART. Courses are available for all assisted conception centre staff including medical, scientific personnel. The unique one-on-one training makes the courses flexible in the course content and the timing to meet your specific needs. Apart from the above, EART also offers other services such as: Turn key projects for IUI, IVF and ICSI Labs. Calibration Services. Quality Management System Implementation. Accreditations Embryology Academy for Research & Training (EART) is a recognized research institute for the purpose of providing the means of instruction and research to the M.V.Sc and Ph.D studies of the Maharashtra Animal and Fisheries Sciences University, MAFSU. EART is also collaborated with www.FertAid.com, Internet Tools http://www.shivaniscientific.com/ivftraining/index.htm[5/13/2009 1:51:18 PM]
  6. 6. Shivani Scientific Industries (P) Ltd. for the Reproductive Sciences, Australia. For more details kindly visit www.ivftraining.com http://www.shivaniscientific.com/ivftraining/index.htm[5/13/2009 1:51:18 PM]
  7. 7. Shivani Scientific Industries (P) Ltd. IVF Products » Mobile Nest Sperm Fuge Lab + Guard Warming Blocks Turnkey Projects Rapid Recovery System IVF Division Concept Human oocytes are extremely sensitive to transient cooling 'IN VITRO' and modest reductions in Temperature can cause irreversible disruption of the meiotic spindle, with possible chromosome dispersal. Embryos subjected to such heat shocks have high proportion of chromosomal abnormalities and this may contribute to the high rates of pre-clinical and spontaneous abortion. Therefore, temperature control is of paramount importance and should not be compromised in any way right from Ovum Pick Up (OPU) till Embryo Transfer (ET). Design This is a handheld device with Li-ion rechargable battery. It is equipped with heating element, Digital sensor, microcontroller, non-volatile memory and LED indicators. When placed on base station, it gets conected electrically by its contacts. These contacts help base station and the mobile device communicates with each other and also charge the battery. The mobile nest when in mobile mode continuously monitors the temperature with a 14 bit accuracy, corrects it several times a second. This yields a rock steady temperature profile even in varying surroundings. The http://www.shivaniscientific.com/ivfproducts/mobilenest.htm[5/13/2009 1:51:46 PM]
  8. 8. Shivani Scientific Industries (P) Ltd. mobile nest continues to log for the power consumption index and temperature abnormalities. These are downloaded to the base station when the nest is docked. With monitoring and reporting features the user can be assured about the mobile nest working accurately at every step. A transparent window facilitates easy viewing of the follicular fluid. How Safe are Preheated Tubes? The Falcon Tubes (2001) pre heated by regular Test Tube Warmer loses its temperature in less than 15 seconds the moment it is out of the heating block and the tube reaches near to the room temperature, thus increasing the risk of thermal shock to the oocytes during aspiration. Mobile Nest is Portable Test Tube Warmer with it's own Internal rechargeable Battery which gives immense mobility and at the same time maintains precise temperature. Intelligent Control System State of the art microcontroller with optomagnetic encoder provides intelligent control system. Digital programming via microprocessor control for various parameters including temperature and power status. The parameters are displayed on four line LCD display Unique Portability This Tubenest has been designed keeping in mind its portable nature. The rechargeable battery is capable of running for 30 minutes at a stretch before it indicates low battery level. The unique locking mechanism for tube allows Embryologist to pour the aspirated follicular fluid into petridish for oocyte scanning very easily. Easy grip design allows convenient handling and battery time facilitates free movement in the LAB and OT. Oprating Security To ensure maximum oocyte survival without quality compromise, this unit has various safety features. The Nest Tube has RED LED INDICATOR which flashes when http://www.shivaniscientific.com/ivfproducts/mobilenest.htm[5/13/2009 1:51:46 PM]
  9. 9. Shivani Scientific Industries (P) Ltd. battery is weak. When the Nest is replaced in the Base Station, it checks the log and indicates any overshoots. Moreover the Nest has a safety over temperature cut out. http://www.shivaniscientific.com/ivfproducts/mobilenest.htm[5/13/2009 1:51:46 PM]
  10. 10. Shivani Scientific Industries (P) Ltd. IVF Products » Lab + Guard Sperm Fuge Mobile Nest Warming Blocks Turnkey Projects Rapid Recovery System IVF Division Concept The LAB + GUARD is developed to improve the laboratory air quality making it nearly equal to clean air. Thus replenishing and subsequently nourishing the entire laboratory, making it safe for the sensitive procedures to be executed. This is achieved by a number of inclusions and their applications all of which are simple, cost effective and user friendly. The positive pressure system uses the basic principle of quot;Over Pressure or positive pressure in a room which restricts the entry of outside air quot;. This is achieved by absorbing the circulating air outside the IVF Lab (Room) and then subsequently passing the same through a series of 4 stage filters thus making it quite IVF friendly. Thus ultimately the effective, circulating air in the IVF lab reaches a near clean air status, provides a positive pressure, reduces the CFU count, eliminates the SPM, VOCs and CACs. Ensure Net Flow Out In any HVAC / air flow design it is essential that net flow is going out of the room. This means room air volume and pressure should be slightly higher than adjoining areas / rooms. This will keep out any toxic fumes trying to come in. IVF labs which are not having adequate positive pressure are suceptible to risk of contamination. Design The LAB + GUARD has been designed with State of the Art technology using 4 stage filtration system. The unique filter combination filters out air and gas contaminant thus by giving most suitable air for gametes and embryos. LAB + GUARD system contains a unique 4 stage filtration system containing Prefilter, Hepa Filter and unique blend of activated carbon and alumina, which ensures wide variety of Gases, particular contaminants are oxidized or are absorbed. The LAB + GUARD has been designed to reduce the microbial air flora to the minimum level. Reduces CFU, Reduces VOC The unique design and the concept of LAB + GUARD positive pressure system not only reduces VOC content in the air but also the CFU count in the IVF Lab. The http://www.shivaniscientific.com/ivfproducts/labplusguard.htm[5/13/2009 1:52:02 PM]
  11. 11. Shivani Scientific Industries (P) Ltd. LAB + GUARD not only protects embryos and gammas but also protects embryologists who is the most affected otherwise. Operating Security Every care has been taken in designing to make sure that air absorbed is thoroughly cleaned for contaminants and to reduce bacterial count in IVF Lab. The LAB + GUARD design also facilitates introduction of fresh air into the air circuit which gives maximum safety from Carbon dioxide released into the IVF Lab by CO 2 Incubators and also it protects embryologists from gas mixtures released on work stations. Change Filters As Per Usage The LAB + GUARD also houses a unique automatic indication system whereby the user is informed when to change the filters, and not quot;every quarterlyquot;. This feature not only gives tremendous relief to the user for tracking the stipulated time of change but also saves lot of money. Maintenance The LAB + GUARD has automatic indication for Filter change and the design facilitate extremely easy replacement of filters even by non technical staff. Other than filter change it is virtually maintenance free Return Blower The return blower is utilised to circulate back the air of IVF lab to room where LAB + GUARD has been fitted. Upgrading, Expanding Or Setting Up New IVF Lab? The LAB + GUARD team helps you in evaluation of possibilities to improve air quality in your lab. If you are upgrading, expanding or setting up new lab contact us for technical assistance. http://www.shivaniscientific.com/ivfproducts/labplusguard.htm[5/13/2009 1:52:02 PM]
  12. 12. Shivani Scientific Industries (P) Ltd. http://www.shivaniscientific.com/ivfproducts/labplusguard.htm[5/13/2009 1:52:02 PM]
  13. 13. Shivani Scientific Industries (P) Ltd. IVF Products » Warming Blocks Sperm Fuge Mobile Nest Lab + Guard Turnkey Projects Are you giving thermal shocks to gametes and embryos? Rapid Recovery Sudden and drastic temperature losses, recovery times and change in pH are System minimized when using Fornax Warming Blocks. The unique aluminum alloy conducts IVF Division heat and provides almost direct thermal contact with the heating surface. Dishes at room temp when kept for incubation without the Warming Block take a prolonged time to attain the set temperature compared to those dishes kept in preincubated Warming Block, also ensures a safe and easy handling of dishes and test tubes in IVF laboratory. GB-4WDRA Made from Aluminum Alloy with hard anodised surface. Holding capacity 4 well NUNC 176740 Dimensions 76 x 98 x 15 mm Weight 112gm. GB-PDRA Made from Aluminum Alloy with hard anodised of surface. Holding One PetriDish Falcon 3002 / capacity 3037 Dimensions 76 x 98 x 15 mm Weight 160 gm. GB-717CA Made from Aluminum Alloy with hard anodised surface. Holding capacity 7 tubes of 14 ml / 17 mm Dimensions 83 x 80 mm(d x h) Weight 888 gm. GB-IDHA Made from Aluminum Alloy with hard anodised surface. Holding capacity One ICSI dish - Falcom 1006 Dimensions 83 x 8 mm (d x h) http://www.shivaniscientific.com/ivfproducts/warmingguard.htm[5/13/2009 1:52:10 PM]
  14. 14. Shivani Scientific Industries (P) Ltd. Weight 44 gm. http://www.shivaniscientific.com/ivfproducts/warmingguard.htm[5/13/2009 1:52:10 PM]
  15. 15. Shivani Scientific Industries (P) Ltd. IVF Products » Turnkey Projects Sperm Fuge Mobile Nest Lab + Guard Warming Blocks Rapid Recovery System IVF Division Shivani Scientific Industries (P) Ltd has vast experience of more than 20 years of designing and building pharmaceutical Laboratories as per WHO-GMP norms with stringent requirement of sterility. The same experience has helped us to fulfill the demanding needs of to-days IVF Labs. A new Lab means start contacting designers, architects, sellers, resellers, vendors, suppliers, agents, distributors to initiate slow, long and laborious process of discussing dates and negotiating rates. This, you will agree, will further eat into your most precious commodity quot;Timequot;. Alternatively you have the easier and simpler option: Contact US. http://www.shivaniscientific.com/ivfproducts/turnkeyproducts.htm[5/13/2009 1:52:21 PM]
  16. 16. Shivani Scientific Industries (P) Ltd. 3D Lab design / 3 D Walk through We offer State of the Art computer modeling to visualize your Lab either in 3D or 3D Walk through, even before your lab is ready. Customised Video Processing All modern equipments comes with all provision of image processing which is very well utilized by our Combination of hardware camera selection and software selection. Multiple Camera inputs can be synthesised in one system to have better recording and processing. Embryology Services We provide assistance in providing complete embryology services from Ovum Pick- up to embryo transfer. Qualified and experienced team of clinicians and embryologists are selected to provide these services. Air Quality evaluation and improvement We have capability to thoroughly evaluate Air quality on existing Lab and recommend appropriate steps to improve the same. It varies from HVAC inspection to microbial air sampling. Hands on training for Lab Staff We offers comprehensive hands on training in collaboration with Maharashtra Animal & Fishery Sciences University to Lab Staff. Annual Maintenance of IVF Lab Equipments We undertake Annual Maintenance Contracts for IVF Labs with the support of its highly qualified team of Engineers. We also cater to calibration contracts with our highly accurate calibration equipments. http://www.shivaniscientific.com/ivfproducts/turnkeyproducts.htm[5/13/2009 1:52:21 PM]
  17. 17. Shivani Scientific Industries (P) Ltd. IVF Products » Rapid Recovery System Sperm Fuge Mobile Nest Lab + Guard Warming Blocks Turnkey Projects IVF Division What is Heat Transfer? To define Heat Transfer requires 2 important group of words. It is a form of energy and in that context we are concerned with the transport of heat energy from one body to another. Motivation for this transfer is provided by the difference in temperature. The definition of Heat Transfer would therefore include a word; transport of energy due to temperature difference. Conduction vs. Convection The word Convection means to bring in or to carry. In other words in normal CO2 Incubator the heat energy is carried through the medium of air to the Petridish, 4 well dish and thereafter it is transmitted to culture media and then to embryos. Whereas Conduction means transfer of heat energy through direct contact of solid substances. In this case the heat energy in the incubator all the heated shelf can be directly transmitted to the 4 well dish or Petridish if they are in contact. Thermal Conductivity Thermal Conductivity of air is 0.034 whereas Thermal Conductivity of Aluminium is 204 and Thermal Conductivity of Stainless Steel is 19. Temperature changes during embryo culture when the dishes are taken out from 37º C incubator into lab where the temperature is anywhere between 24º C to 28º http://www.shivaniscientific.com/ivfproducts/rapid-recovery-system.htm[5/13/2009 1:52:30 PM]
  18. 18. Shivani Scientific Industries (P) Ltd. C and also when culture media which are used in embryo culture lose the temperature when they are droplet from flat surface. Temperature changes recovery time and PA changes are drastically reduced when Rapid Recovery System is used with regular CO2 Incubator. The new Rapid Recovery System provides direct contact, which promotes heat further through conduction to bottom of the culture dish. Loss of temperature on heated surfaces The air gap under culture dishes promotes bad conduction between incubator shelf and the bottom of the dishes and as air is very bad conductor of heat it is quite deter mental to the embryos and gametes. Recovery of CO2 Incubator When the dishes are kept back into the CO2 Incubator, CO2 often shows recovery of the temperature on the display. However, it does not mean the temperature of Embryos and Oocyte has reached. It is often reported that the actual stabilization on 37º C in the Embryo happens after 22-30 minutes of Incubator. The New Rapid Recovery System The New Rapid Recovery system has been designed to enhance Embryo development and the culture conditions by recovering temperature practically faster than conventional methods. The new Rapid Recovery System is to be used with our GB Series Warming Blocks. The Rapid Recovery System has necessary design, feature to accommodate 4 well dishes, Nunc Dishes and Falcon Petridishes. The Rapid Recovery system give you direct thermal contact between the dry block type heating system and the individual culture dish wells, ensuring rapid temperature equilibration combined with low temperature variability. http://www.shivaniscientific.com/ivfproducts/rapid-recovery-system.htm[5/13/2009 1:52:30 PM]
  19. 19. Shivani Scientific Industries (P) Ltd. IVF Products » IVF Division Sperm Fuge Click here to know more Click here to know more Mobile Nest Lab + Guard Warming Blocks Click here to know more TurnKey Projects Click here to know more Rapid Recovery System Click here to know more Click here to know more Click here to know more Click here to know more Click here to know more Click here to know more Click here to know more Click here to know more Click here to know more Click here to know more http://www.shivaniscientific.com/ivfproducts/ivfproducts/index.htm[5/13/2009 1:52:44 PM]
  20. 20. Temperature Controlled Centrifugation: - The Effect on Motion Characteristics of Human Sperm Dr Franken, K Avari Department of Obstetrics & Gynaecology, University of Stellenbosch, Tygerberg Hospital, Tygerberg, 7505, South Africa February 2007 Introduction Material & Methods Semen samples and sperm preparation Sperm retrieval techniques form an integral part of the assisted reproductive programme. Semen samples were obtained from The success of sperm separation is measured normozoospermic donors and andrology by the number of motile sperm retrieved from a referrals. Directly after collection samples were given semen sample. The influence of various divided into 2 aliquots. One aliquot were placed in a CO2 incubator at 340C while the technical procedures and diluent media upon human spermatozoa has been tested in vitro second aliquot were left at room temperature. (Jeulin et al., 1982). The percentage and A period of 30 minutes was allowed to stabilize velocity of motile spermatozoa are often the semen temperatures before experimental measured objectively using laser Doppler procedure. Prior to the experimental onset the velocimetry or Hamilton Thorne IVOS 10 motion characteristics were recorded with the computerized sperm analyzer (CASA). When CASA instrument in order to obtain initial spermatozoa are incubated at 37 degrees C, values for the measured motion rapid declines in both percentage motile and in characteristics. velocity were observed with incubation periods lasting more than 4-h (Jeulin et al., 1982). Swim up separation Human sperm incubation at room temperature Motile sperm fractions were retrieved from the does not allow capacitation, although it does semen samples by mixing 0.5mL semen with not affect hFF-induced acrosome reaction in 1mL of Quinn’s Sperm Preparation medium. capacitated cells. The blocking effect is Two tubes were prepared for each experiment. overcome when spermatozoa are exposed to The tubes were then placed in 2 different 37°C (Maryn-Briggiler et al., 2002). Keppler et centrifuges, namely (i) SpermFuge SF 800 al., (2002) found significantly higher percent (Shivani Industries, Mumbai, India), a highly motility, mean average path velocity, straight precise centrifuge with a temperature line velocity, lateral head displacement, and controlled chamber at 340C and (ii) Sigma percent hyperactivation in sperm at the 40ºC bench top with no temperature control temperature (Keppler et al., 1999) facilities. Both centrifuges were set at 1500rpm (428xG) for 5 minutes. Following the second washing procedure, both sperm Objectives pellets were layered with culture medium and left at a 450 angle at 340C for 60 minutes. After the incubation period 0.5mL supernatant were To evaluate the effect of temperature on the removed from each tube and immediately sperm motion characteristics and subsequent analyzed for sperm motion characteristics. outcome of sperm preparation. 1
  21. 21. Temperature Controlled Centrifugation The following parameters were evaluated: Motion characteristics sperm concentration; motile and progressively Aliquots of 5µL of spermatozoa (10x106 motile concentrations; percentage motile and cells/mL) from the sperm at 34ºC and 22ºC progressively motile; path velocity (VAP); aliquots were placed in the micro chamber for straight-line velocity (VSL); curvilinear velocity analyses with the HTM-IVOS V10.9 CASA (VCL); amplitude of lateral head displacement instrument (Hamilton-Thorne Research Inc., (ALH); beat cross frequency (BCF); Beverley, MA, USA). The following standard straightness (STR); and linearity (LIN). Motion set-up parameters: 30 frames/60 Hz; minimum characteristics were recorded in all samples contrast, 80; minimum cell size, 2; minimum using 10 randomly selected microscopic fields. static contrast, 30; low VAP cut-off, 5 µm/s; The proportion of hyperactivated spermatozoa low VSL cut-off, 11 µm/s; head size, non- in each experiment was determined using the motile, 3; head intensity, non-motile, 160; SORT function of the CASA instrument. To be static head size, 1.01-2.91; static head classified as hyperactivated, a trajectory had to intensity, 0.60-1.40; slow cells, non-motile; meet all of the 60 Hz SORT criteria, i.e., magnification, 2.01; and temperature, at 34ºC. VCL≥150µm/s, LIN≤50% and ALH≥7µm (Mortimer et al., 1990). Results Table 1 Results of semen parameters recorded in ejaculates and after preparation with SpermFuge temperature controlled vs. room temperature centrifugation Post Swim up Ejaculates 340C Spermfuge Room temperature Volume Morphology Motile Sperm Sperm Sperm % normal Concentration concentration Concentration mL % live Mill/mL Mill/mL Mill/mL Mean 3.20 10.80 58.00 80.60 12.10 a 7.30 b SD 1.15 3.04 24.40 53.14 6.63 2.38 a vs.b Fisher’s exact test p = <0.05 Significant difference obtained in post swim up sperm concentrations Table 2 Percentage Hyperactivated sperm recorded after preparation with SpermFuge vs. Room temperature prepared % Hyperactivated sperm BASELINE SWIMUP 34OC 20OC 34OC 20OC 0.8% 0.8% 1.73% a 0.35% b Transformed ARCSIN values showed a vs.b Fisher’s exact test p=<0.05 Significant difference obtained in percentage hyperactivated sperm 2
  22. 22. Temperature Controlled Centrifugation Table 3 Comparison of the velocity distribution of sperm cells( %) prepared by a SpermFuge temperature controlled (34oC) centrifuge vs. room temperature swim up spermatozoa SPERM VELOCITIES DISTRIBUTED IN 5 MOTION CATEGORIES % RAPID % MEDIUM % SLOW % STATIC % HA Baseline Baseline Baseline Baseline Baseline 34 C 20 C 34 C 20 C 34 C 20 C 34 C 20 C 34 C 20 C o o o o o o o o o o Mean 4.2 4.5 4.8 3.2 3.5 3.6 4.0 3.6 3.3 4.0 3.6 3.9 0.4a 1.2b 0.3c SD 0.7 0.7 1.3 0.5 0.7 0.6 0.5 0.6 1.0 0.7 0.4 0.7 0.8 1.2 0.8 Transformed ARCSIN values showed a vs.b, a vs c and b vs. c Fisher’s exact test p=<0.05 Significant difference obtained in percentage hyperactivated sperm References 1. Mortimer, D. (1990) Objective analysis of sperm motility and kinematics. In Keel B.A. and Webster, B.W. (eds), Handbook of the Laboratory Diagnosis and Treatment of Infertility. CRC Press, Boca Raton, pp. 97-133. 2. Holt WV and Palomo MJ (1995) Optimization of a continuous real-time computerized semen analysis system for ram sperm motility assessment, and evaluation of four methods of semen preparation, Reproduction, Fertility and Development 8(2) 219 - 230 3. Jeulin C, Serres C, Jouannet P. (1982) The effects of centrifugation, various synthetic media and temperature on the motility and vitality Reprod Nutr Dev,22(1A):81-91 4. Makler A., Deutch M., Vilensky A., Palti Y. (1981) Factors affecting sperm motility. VIII. Velocity and survival of human spermatozoa as related to temperatures above zero International Journal of Andrology, 4 (5), 559-569. 5. Maryn-Briggiler CI, Tezon JG, Miranda PV, Vazquez-Levin MH. (2002) Effect of incubating human sperm at room temperature on capacitation-related events. Fertil Steril 77, (2) 252-256. 6. Keppler EL, Chan PJ, Patton WC, King A. (1999) Aggregation of human sperm at higher temperature is due to hyperactivation. Arch Androl 42; 35-39. DFR22022007-F3 3

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