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RMG/ART Program and IVF Laboratory Information

The Reproductive Medicine Group is a leader in the treatment of Infertility in the Tampa Bay region. Assisted reproductive technology (ART) therapies, the most common of which is in vitro fertilization (IVF), assist countless individuals and couples in overcoming fertility issues through the joining of the sperm and eggs outside the body and then transferring the resulting embryo into the uterus. The RMG/ART program is comprised of dedicated physicians, nurses, embryologists, and andrologists. Our ART Laboratory is located adjacent to our Surgery Center in North Tampa.

About the RMG/ART Laboratory

The RMG/ART Laboratory is fully accredited by the College of American Pathologists (CAP) in the areas of andrology and embryology. This accreditation indicates that the RMG/ART Laboratory belongs to an exclusive group of laboratories throughout the country that meet the highest standards of excellence as recognized by their peers. The accreditation process includes an evaluation of all aspects of quality in the laboratory:

  • Methodology
  • Reagents
  • Medical equipment control
  • Specimen handling
  • Procedure manuals
  • Reports and proficiency testing
  • Personnel
  • Overall management principles that distinguish a quality laboratory

The CAP is an advocate for high-quality and cost-effective medical care.

The RMG/ART Laboratory also participates in the biannual proficiency testing by The American Association of Bioanalysts. This comprehensive external quality control program satisfies the proficiency testing requirements for Clinical Improvement Amendments, the Commission on Office Laboratory Accreditation, Health Care Financing Administration, the Joint Commission on Accreditation of Healthcare Organizations, CAP, and most state agencies.

The Reproductive Medicine Group ART Laboratories Inc. in Tampa, FL has been awarded accreditation by the Accreditation Committee of the College of American Pathologists (CAP), based on the results of a recent onsite inspection in December 2010. The Reproductive Medicine Group was advised of this national recognition and congratulated for the excellence of the services being provided.

RMG/ART Laboratories Inc. is one of more than 7,000 CAP-accredited laboratories worldwide. The CAP Laboratory Accreditation Program, begun in the early 1960s, is recognized by the federal government as being equal to or more stringent than the government’s own inspection program. This stringent inspection program is designed to specifically ensure the highest standard of care for all of our patients.

How does the art laboratory operate and what safety measures are taken? What will happen to the eggs and sperm after retrieval?

On the day before the oocyte (egg) retrieval, oocyte culture dishes are labeled with the patient’s name, date of birth, and ART identification number. The number and size of the ovarian follicles determine the number of dishes prepared (one dish for each large follicle). The dishes are filled with a rinsing fluid in the outer well for removing blood and other extraneous cells from the egg, and a nutrient fluid for the culture of the egg is in the center well.

The dishes are then placed in the embryology incubator for warming to body temperature. The incubator also adjusts the pH of the culture media to the human body. A large amount of rinsing media is equilibrated in the incubator. Media is also equilibrated in the andrology laboratory for the processing of sperm.

One of the more important and useful IVF laboratory instruments is the mobile IVF chamber. It is a pediatric isolette (incubator) on wheels that has been modified to contain a scanning microscope. Prior to the egg retrieval, it is warmed to body temperature and put at the normal neutral pH of the culture dish media. These conditions avoid any abrupt changes the eggs might encounter outside the body. The chamber has two small doors on each side. The embryologists place their hands on one side to identify eggs and move dishes. The OR nurse uses the opposite doors to pass test tubes containing the follicular fluid to the embryologist.

Upon arrival in the operating room, the patient’s identity is verified by the embryologist, physician, and safety nurse. They also verify that the culture dishes are correctly labeled. At the time of surgery, the mobile chamber is loaded with equilibrated petri dishes for receiving the follicular fluids.

The physician aspirates the fluid from the ovarian follicles into a test tube. A nurse passes the test tube, through the IVF isolette chamber door, to the embryologist. The embryologist then dispenses the fluid into a large dish and rapidly scans it under the microscope. The egg complex is usually visible to the naked eye, but microscopic verification is always done. The oocyte is removed from the large dish by pipette, rinsed in the outer well of the culture dish, and placed in the center well for culture and the addition of sperm (insemination). The presence of an egg in the follicular fluid is reported to the physician.

After all of the eggs have been aspirated, the mobile chamber is moved to the embryology laboratory for a more critical assessment of the oocytes, insemination with sperm, and culture.

The maturity and health of oocytes are assessed after an incubation period. Oocytes are graded as mature, intermediate, immature, or degenerate. Oocytes judged mature are incubated for 1 to 4 hours before insemination. Less mature oocytes are incubated until they reach maturity, and then inseminated. Approximately 100,000 motile sperm are added to the media surrounding the oocyte in the center well of the culture dish. Dishes containing egg and sperm are then returned to the main embryology incubators for culture.

The male will be asked to collect a semen sample after the oocyte aspiration. The andrologist escorting him to the collection room will identify the patient’s partner from his identification bracelet prior to collection and will write both names on the outside of the specimen container after collection. The partner will be asked to verify that both names are correct.

Approximately 30 minutes after collection, when the semen liquefies, a semen analysis is performed. The sperm is prepared for insemination.

About 20-24 hours after retrieval, the oocytes are transferred to fresh dishes that were equilibrated on the day of retrieval. Oocytes are then microscopically examined for pronuclei formation, the sign of fertilization. The pronuclei are the decondensed DNA of sperm and egg nuclei. They appear microscopically as two light spheres within the egg. Oocytes possessing more than two pronuclei are discarded.

Fertilized oocytes are returned to the main embryology incubator for 24 hours. At that time the embryos are transferred to fresh dishes where the cell stage and overall health of the embryos are microscopically evaluated. The embryos are either transferred on day 3 or placed into new dishes with a specialized culture media to allow for progression to the blastocyst stage.