Jane E. Johnson

Control of air quality in an assisted reproductive technology laboratory.

OBJECTIVE To investigate the effect of improved air quality on IVF and subsequent embryo development. DESIGN Retrospective cohort study. SETTING Hospital-based IVF facility composed of an anteroom, a cleanroom, and an adjacent operating room. PATIENT(S) Two-hundred seventy-five couples requesting IVF between 1993 and 1997. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Particle counts (sizes 0.3, 0.5, 1.0, and 5.0 microm); IVF rates; and embryo quality (stage and grade). RESULT(S) Clinical pregnancy rates decreased from 35% in 1993 to 16% in 1994 (numerous construction odors were detected during 1994) and increased steadily after the cleanroom was built (rates for 1995-1997 were 20%, 32%, and 59%, respectively). Fertilization rates decreased between 1993 (74%) and 1994 (60%) and then steadily increased after cleanroom installation (62% in 1995, 71% in 1996, and 69% in 1997). The proportion of embryos past the four-cell stage decreased from 66% in 1993 to 61% in 1994 but then increased steadily in the years after the cleanroom was built (78%, 77%, and 83% in 1995, 1996, and 1997, respectively). During the same 5-year period, there were no differences in embryo quality or number of embryos transferred. CONCLUSION(S) Construction of a Class 100 cleanroom improved air quality and IVF rate and increased the number of embryos past the four-cell stage available for transfer.

Manual versus computer-automated semen analyses. Part I. Comparison of counting chambers

OBJECTIVE To determine the accuracy and precision of counting chambers analyzed manually and with a computer-automated semen analyzer (CASA; Hamilton-Thorne Research, Beverly, MA). DESIGN Prospective study using comparative measurements of sperm concentration, motility and concentration of latex beads with three types of counting chambers: hemacytometers, 12 and 20 MicroCell (Conception Technologies, Inc., La Jolla, CA) Chambers, and Makler (Sefi-Medical Instruments, Haifa, Israel) Chambers. SETTING A hospital-based Andrology laboratory. PATIENTS Male partners of couples undergoing infertility evaluation. MAIN OUTCOME MEASURES Experiment I: measurements of sperm concentration were evaluated within hemacytometers; Experiment II: measurements of sperm concentration in the 35 to 50 x 10(6)/mL range and sperm motility were determined using a known concentration of beads. RESULTS Experiment I: differences were demonstrated within two of eight hemacytometers (side 1 versus side 2): however, no significant effect of hemacytometer on variation in sperm concentration measurements was observed. Experiment II: CASA-analyzed 20 MicroCell Chambers demonstrated the best precision for sperm concentration (intraclass correlation coefficient = 0.93) and motility (intraclass correlation coefficient = 0.88). Experiment III: 20 MicroCell Chambers most accurately determined known bead concentration (35 +/- 5 x 10(6)/mL) whether analyzed on CASA (34.9 x 10(6)/mL) or manually (35.2 x 10(6)/mL). CONCLUSIONS 20 MicroCell Chambers proved to be accurate and precise for determining concentration and motility of semen specimens whether analyzed manually or with CASA.

Short Communication: Cook Versus EdwardsWallace: Are There Differences in Flexible Catheters?

The most important 10 min in an assisted reproductive technology (ART) procedure may be the last 10 min—the embryo transfer. Bongso suggests that pregnancy rates for in vitro production (IVP) clinics are determined by proper stimulation, culturing techniques, and atraumatic embryo transfers (1). Focusing on the latter, (a) we compare our clinical pregnancy rates after embryo transfer with the Cook softpass catheter and the Edwards–Wallace catheter, and (b) because two other independent studies with the same research question report similar findings (2,3), we combine our data with theirs to compare these two popular embryo transfer catheters.

Manual versus computer-automated semen analyses. Part III. Comparison of old versus new design MicroCell Chambers

OBJECTIVE To determine accuracy and precision of old and new design MicroCell counting Chambers analyzed manually and with a computer-automated semen analyzer (CASA; Hamilton-Thorn Research, Beverly, MA). DESIGN Prospective study using comparative measurements of the concentration of latex beads with old and new design MicroCell Counting Chambers (Conception Technologies, Inc. La Jolla, CA). MAIN OUTCOME MEASURE Beads were counted manually and with CASA to evaluate the accuracy of old and new design 20 MicroCell Chambers. RESULTS The old design 20 MicroCell Chamber demonstrated a significant difference in bead concentration beginning at the 9.9 mm CASA stage position compared with stage positions of 0.0 to 9.5 mm. The new design 20 MicroCell Chamber demonstrated the same difference at the 8.0 mm CASA stage position compared with stage positions 0.0 to 7.6 mm. Similar findings were observed when the beads were analyzed manually. CONCLUSIONS When analyzed within a specific range of stage positions, the old and new design 20 MicroCell Chambers are accurate and precise whether analyzed manually or with CASA.

Can Westgard Quality Control Rules Determine the Suitability of Frozen Sperm Pellets as a Control Material for Computer Assisted Semen Analyzers

AbstractPurpose: To evaluate the drop-to-drop and pellet-to-pellet repeatability and stability of frozen sperm pellets. Methods: Ten pellets were thawed per batch (low and normal concentration) and evaluated by two investigators to establish a quality control chart. Then low and normal concentration pellets were thawed and evaluated daily for 10 days by both investigators. The values for both investigators were averaged and plotted on the chart. Results: The low sperm concentration specimen had a systematic error while the normal sperm concentration specimen had a random error as well as a systematic error. The low sperm concentration specimen violated the warning rule for motility whereas the normal concentration violated the warning rule, the random error rule, and the systematic error rule when applied to motility. Conclusions: Frozen sperm pellets are not acceptable as a daily-use quality control material for semen analysis when using a computer assisted semen analyzer.

Short Communication: Can Varying the Number of Spermatozoa Used for Insemination Improve In Vitro Fertilizaton Rates?

growth from secondary stages only (1). Therefore, PETAH TIKVA, ISRAEL before clinical use, the growth and maturation capacity of frozen-thawed fetal oocytes need to be tested. The Development of Human Fetal Follicles in aim of the present study was to examine the ability of an Immunodeficient Mouse* human fetal oocytes to develop in an immunodeficient nu/nu mouse and to assess their growth by immunocytochemical expression of proliferating cell nuclear antiSubmitted: November, 24, 1999 gen (PCNA). PCNA is a protein involved in cell cycle Accepted: January 29, 2000 found to correlate with in initiation of human adult (2), bovine fetal (3), and baboon (4) folliculogenesis and not expressed in primordial follicles. INTRODUCTION

Quality Management Issues in the Assisted Reproduction Laboratory

In the United States, the Clinical Laboratory Improvement Act (CLIA) of 1988 describes requirements and guidelines for implementing a quality control/quality assurance (QC/QA) program for moderate ...

Clinical Assisted Reproduction: Extending the Coincubation Time of Gametes Improves In Vitro Fertilization

Altering the coincubation time for spermatozoa and oocytes in vitro leads to varying results. Some authors suggest that longer exposure of spermatozoa to oocytes through coincubation is detrimental to fertilization and/or embryo development, possibly because of reactive oxygen species produced by the spermatozoa (1–5). Other studies indicate that a decreased exposure time does not alter fertilization rates (3,6–9) and may even improve such rates (10–13). In this report, we describe the results of our randomized study on the effects of two coincubation times on in vitro fertilization (IVF) rates and embryo quality.

Using Fyrite to monitor incubator carbon dioxide levels

PurposeThe Fyrite analyzer (Bacharach, Incorporated) was evaluated to ascertain whether it would perform within the range specified by the manufacturer (±0.5%). Fyrite analyzer samples were compared to control samples obtained using an infrared CO2monitor (Ohmeda, Model Number 5200).MethodsTen Fyrite analyzer measurements from each of three incubators were obtained using two different analyzers by three investigators for a total of 60 measurements per investigator. Twenty control measurements for each of the three incubators were obtained using an infrared monitor calibrated at the 5.0% CO2level.ResultsFyrite analyzer values fell outside of an acceptable range (Control mean CO2±0.5%) for 7 of 60 (11.7%) observations for Incubator 1 and 4 of 60 (6.7%) observations for Incubator 2. (Incubator 3 had no values outside the range.) The Fyrite analyzers were significantly different from each other (P=0.002). The three investigators (I) differed significantly from the control (I1 and I3, P<0.001; I2, P=0.02) as well as from each other (I1 vs I2, P<0.001; I1 vs I3, P<0.001; I2 vs I3, P<0.016).ConclusionsUse of an infrared monitor for incubator calibration and monthly CO2checks in concert with daily use of the Fyrite as a monitoring tool only is the better method.

The effect of the culture vessel and insemination method on the in vitro fertilization and development of human oocytes

ConclusionsOur laboratory has corroborated previously published work demonstrating that tissue culture tubes and microdrops perform equally well for in vitro fertilization and culture of human oocytes and embryos.