PubMed · 4600739
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P M Messier. 1974. [Scanning electron microscopy].. https://pubmed.ncbi.nlm.nih.gov/4600739/
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OBJECTIVES: To correlate the percentage normal morphology reported by different observers and a computerized method (IVOS; Hamilton-Thorne Research, Beverly, MA) on a slide-by-slide basis using strict criteria: [1] Experienced observer (T.F.K.) versus experienced observer (R.M.), [2] experienced observer (T.F.K.) versus sperm morphology analyzer (IVOS), and [3] repeatability of normal and abnormal cells (IVOS). DESIGN SETTING, PATIENTS: Slides from 30 different patients from the Tygerberg IVF program were selected randomly. Microscopic fields and sperm cells were chosen randomly and percent normal morphology was recorded (objectives 1 and 2). The same slides were used and a cell-by-cell repeatability was done as outlined (objective 3). RESULTS: Experiment 1 (objective 1): there was no significant bias between T.F.K. and R.M. The limits of agreement were 8.6% and -7.3%. The SDs were not significantly different (P = 0.1283). The Spearman correlation coefficient between readers was 0.83. Experiment 2 (objective 2): the same findings were reported but the limits of agreement were 12.1% and -15.5%. The Spearman correlation coefficient was 0.85. The limits of agreement was tighter below 20% normal forms (+8.4 and -6.6). Experiment 3 (objective 3) (repeatability): 255 cells were analyzed three times in succession. Estimating pairwise agreement, the kappa statistic for the pairs are 0.85, 0.80, and 0.85, respectively, which compares favorably with the second canonical moment of 0.8329 (kappa = 0.83). DISCUSSION: The computer's ability to classify normal morphology per slide is promising. Below 20% normal forms, the limit of agreement is tight. Because of the 6% higher reading compared with the manual method, different thresholds possibly will be developed to identify subfertile from fertile patients. The computer gives excellent repeatability of normal and abnormal cells. Based on results obtained, this system can be of clinical value both in IVF units and andrology laboratories but more clinical data is required in this field.
Currently, there are no standard controls available for quality control of DNA ploidy assays in image analysis systems. The authors have developed a system of controls based on cultured human cell lines that can mimic tissue sections and touch preparations. This allows for quality control for the entire process for DNA ploidy analysis from the initial cutting of the paraffin block or production of the touch preparation through fixation, staining, and interactive image analysis. The use of cell line controls rather than normal tissue is advantageous because of cell population homogeneity and the volume of uniform slides that are obtainable. This system, while providing an excellent means of controlling day-to-day performance of DNA ploidy analysis, may also be adapted for use as a means of multi-institutional proficiency testing.
The Clinical Laboratory Improvement Amendments of 1988 (CLIA '88) contain regulation 493.1257(c)(1), which states: "[A]ll gynecologic smears interpreted to be showing reactive or reparative changes are confirmed by a technical supervisor in cytology." We surveyed 500 cytology laboratories to assess the utilization of the diagnosis "reactive/reparative" (R/R) for gynecologic cytology examinations. One hundred and seventy participants (34%) returned an acceptable questionnaire that described their use of this terminology, the technical supervisor's (pathologist's) role in the evaluation of these cases, and the volume of cases with this diagnosis. Prior to CLIA '88, 86 (51%) laboratories did not forward to a pathologist gynecologic smears reported as showing R/R changes, and this decreased to 20 (12%) after implementation of CLIA '88. Thirty-six (21%) laboratories changed their criteria for reporting R/R changes after CLIA '88. No correlation was noted between the demographics of the laboratory (e.g., size, type of practice affiliation, accreditation) and familiarity with this regulation. Based on the case volumes provided in the survey results, the estimated cost to the nation of complying with this single regulation is between $4.5 million and $40 million. The interpretation of and financial impact of compliance with this one selected regulation of CLIA '88 is reported.