Scrape Preparations Research Articles

Utility of Ovarian Scrape Preparations for the Intraoperative Diagnosis of Ovarian Epithelial Lesions

Utility of Ovarian Scrape Preparations for the Intraoperative Diagnosis of Ovarian Epithelial Lesions

Fungal foot abscess caused by Aureobasidium pullulans culture diagnosis of fine needle aspiration cytology material in a clinically unsuspected patient

Subcutaneous mycosis includes a large spectrum of infections caused by a heterogeneous group of fungi. Fungal infections of the skin can be identified in scrape preparations of lesions and Fine Needle aspirates of abscesses, with recognition of the specific morphological features such as hyphae and branching. We report a fungal abscess caused by Aureobasidium pollulans by smears and culturing the material obtained from Fine Needle Aspiration Cytology.

Cytopathologic evaluation of the in situ margin in patients undergoing hepatectomy

Margin status is a predictor of outcome for patients with liver malignancies, although what constitutes a negative margin is controversial. Traditionally, the completeness of resection is estimated by surgical histopathology of the resected specimen margin, despite the in situ margin being potentially more important. The true margin is often altered by parenchymal transection techniques. The authors propose that cytologic assessment of the in situ margin is more specific for determining the true margin. A total of 84 patients with primary or metastatic liver tumors who were undergoing surgical resection were enrolled in this prospective Institutional Review Board-approved study. Specimen and in situ (patient) margins were assessed using a "scrape preparation" cytologic technique and compared with traditional surgical histopathology. Patients were followed for assessment of local disease recurrence. Follow-up data were complete for 64 patients for a median of 37 months (range, 12 months-56 months). Twenty patients were excluded because of perioperative death (6 patients; 7%) or a follow-up of < 12 months. Seven patients (12.2%) had positive histopathologic specimen margins, but only 1 was found to be positive by cytology (1.8%). No in situ cytologically positive margins were identified along the cut edge of the liver remnant. The rate of intra- or extrahepatic recurrences was 56.7%, whereas the local recurrence rate was 1.8%. One patient with local recurrence demonstrated simultaneous intra- and extrahepatic disease recurrences and had negative margins by all methods of evaluation. To the authors' knowledge, the current study is the first to demonstrate that in situ margins can be assessed using cytopathology. This method is quick and can be universally applied. Given the difficulty of accurately assessing margins after hepatectomy, cytopathologic evaluation may be more reflective of the true margin. Cancer (Cancer Cytopathol) 2012. © 2012 American Cancer Society.

Cytopathology of alveolar soft part sarcoma

Alveolar soft part sarcoma (ASPS) rarely is subjected to cytopathologic evaluation. With the exception of some very small series, the literature is limited to case reports. The objectives of the current study were to evaluate the cytomorphology of 10 ASPS cases on fine-needle aspiration (FNA) or imprint cytology, review the literature, and highlight potential diagnostic pitfalls. The authors searched their files for all lesions that were signed out as ASPS or suspicious for ASPS and searched the surgical pathology files for any cases of ASPS that had corresponding cytology. FNA was performed using the standard technique. Scrape preparations were performed on tissues that were sent fresh for frozen section examination. Ten cases of ASPS were retrieved from 7 patients (male-to-female ratio, 4:3; mean age, 22 years). All had subsequent tissue confirmation. Six specimens were from primary lesions, 3 specimens were from metastases, and 1 intraoperative smear was from a previously aspirated primary mass in a newly diagnosed patient. Anatomic sites of involvement included extremities (5 specimens), trunk/pelvis (3 specimens), oral cavity (1 specimen), and lung (1 specimen). Eight specimens were aspirates, and 2 specimens were intraoperative smears. Nine cases were diagnosed correctly as ASPS/consistent with ASPS, and 1 specimen was diagnosed as "tumor, not otherwise specified". Cytomorphology included variably cellular smears composed of large cells with an enormous amount of finely granular or vacuolated cytoplasm, markedly enlarged nuclei/nucleoli, and bare nuclei. The cytomorphology of some cases revealed a strong similarity to renal cell carcinoma, clear cell type. The results of the current study indicated that ASPS has cytomorphology that overlaps with several other neoplasms, including renal cell carcinoma. Nonetheless, the morphologic features, when combined with the clinical presentation, radiologic findings, and ancillary testing, may allow for a specific diagnosis.

Intraoperative cytologic evaluation of sentinel lymph nodes in patients with breast carcinoma by scrape preparation

Intraoperative evaluation of sentinel lymph nodes (SLNs) in patients with breast carcinoma allows surgeons to complete axillary lymph node dissection in one procedure if any SLN shows metastasis. The accuracy of intraoperative pathological diagnosis is critical for decision-making. The purpose of this study was to evaluate our rapid intraoperative cytologic diagnosis of SLN through comparing with the final surgical pathologic diagnosis of the corresponding lymph nodes. A total of 454 SLNs from 159 consecutive female patients with a preoperative diagnosis of breast carcinoma over 3-year period were included in this study. After gross examination of each bisected lymph node, a scrape preparation was prepared for each submitted lymph node and was stained by the rapid Papanicolaou method. The intraoperative cytologic diagnosis was compared with the final surgical pathologic diagnoses. The overall sensitivity of intraoperative cytology was 52.5% with specificity of 100%. There were 17 false-negative cases. Of them, six nodes had isolated tumor cells, seven nodes had micrometastasis (0.2-2 mm), and four nodes had macrometastasis (>2 mm). There were no interpretive errors identified. The size of metastasis and tumor grade appeared to be significant factors in detecting metastasis by cytology. In addition, subsequent non-SLN involvement was 9% in patients with micrometastasis versus 50% in patients with macrometastasis (P < 0.05). Our study shows that the intraoperative cytologic evaluation of SLNs in breast carcinoma is a reasonably accurate method. The majority of false-negative cases were due to micrometastasis and isolated tumor cells.

Sentinel Lymph Nodes

Sentinel Lymph Nodes

Cytopathology of extraskeletal myxoid chondrosarcoma

Extraskeletal myxoid chondrosarcoma (EMC) is a rare soft-tissue sarcoma rarely subjected to cytologic analysis. With the exception of a few small series, the cytology literature of EMC is largely limited to single-case reports. The purpose was to evaluate the cytomorphology of 8 EMC cases as obtained by imprint/scrape cytology and fine-needle aspiration (FNA) biopsy, to review the literature, and to demonstrate the utility of cytogenetic analysis in the diagnosis of EMC. The cytology files were reviewed for all soft-tissue lesions signed out as chondrosarcoma, myxoid sarcoma, and EMC, and the tissue files for any cases of EMC that had corresponding cytopathology. FNA was performed using a standard technique. Scrape preparations were performed from tissue sent fresh to the laboratory for either frozen section or for special studies such as electron microscopy or tissue banking. Eight cases of EMC were retrieved from 4 men and 3 women (median age = 62 years). One patient had 2 separate cytologic specimens 4.5 years apart. All patients had subsequent tissue confirmation of the diagnosis of EMC. Five individuals presented as new patients, and 2 had a prior diagnosis of EMC. Sites included 4 masses from the foot/ankle, 2 from the calf, 1 wrist mass, and 1 buttock mass. Five patients were diagnosed from FNA biopsy, whereas 3 were diagnosed using scrape slides. Five cases were correctly and categorically diagnosed by the cytologic method as EMC, 1 as chondrosarcoma favor EMC, 1 as sarcoma favor EMC, and 1 as myxoid spindle/epithelial neoplasm. Cytologic features ranged from hypocellular to highly cellular smears composed primarily of rounded cells set in an abundant myxoid stroma that varied from opaque to semitransparent and lacked vascularity or necrosis. Smears showed cells in short, sometimes anastomosing cords, but also as single cells and nondescript cell clusters. Cells displayed a monotonous uniformity in nuclear diameter and cell size. Bland nuclei with evenly dispersed chromatin displayed variably sized nucleoli, and a moderate amount of infrequently vacuolated cytoplasm. Tissue fragments of variable size were found in 5 of 5 FNA cell blocks. Fluorescence in situ hybridization (FISH) analysis using the EWSR1 probe showed a positive 22q12 translocation in 2 of 3 FNA cases that were tested. One case with negative FISH results on the cytologic preparation showed a positive translocation using the same technique in the subsequent resection specimen. A confident cytologic diagnosis of EMC depends on the presence of a uniform, round to oval cell population often arranged in cords and set in an abundant myxoid/chondromyxoid background and arising in the appropriate clinical context. If positive, FISH testing (of paraffin cell blocks or cytospin preparations) is confirmatory when coupled with this cytomorphology.

Intraoperative Pathologic Examination: Cost Effectiveness and Clinical Value in Patients with Cytologic Diagnosis of Cellular Follicular Thyroid Lesion

Routine use of intraoperative pathologic examination (IOPE), including frozen section (FS) and scrape preparation cytology (SPC), during diagnostic thyroid lobectomy continues to be a source of controversy. We sought to better delineate the usefulness and cost-benefit ratio of IOPE in the context of cytologically diagnosed cellular follicular lesion (CFL) or follicular neoplasm (FN). Records of 205 patients who underwent thyroidectomy for cytologically diagnosed FN or CFL between 1997 and 2005 were retrospectively reviewed. IOPE results, patient demographics, and tumor characteristics were correlated to final histopathologic diagnoses. Sensitivity, specificity, predictive values, accuracy, and costs of IOPE were calculated. IOPE correctly identified 3 of 16 follicular carcinomas and 9 of 36 papillary carcinomas. Sensitivity, specificity, and accuracy were 23%, 99%, and 78%, respectively. On univariate analysis, malignancy risk among follicular nodules did not correlate with age, gender, or nodule size. On multivariate analysis, nodule size was predictive of malignancy (p < 0.05). Over the entire patient series, routine IOPE resulted in a net cost savings of $74,304.33. IOPE reduced costs and limited the number of completion thyroidectomies necessary. IOPE is specific, cost effective, and of minimal additional risk when performed routinely for patients with CFL or FN.

Sentinel node micrometastasis in breast carcinoma may not be an indication for complete axillary dissection

The decision whether to proceed with complete axillary node dissection based on sentinel node status is clear for patients with negative or macrometastatic disease. However, the course of action based on sentinel node micrometastasis remains controversial. We reviewed 358 cases from 6/1999 to 7/2003. All sentinel nodes were evaluated at three levels by frozen section, touch preparation, and scrape preparation. Micrometastasis was defined as tumor deposits between 0.2 and 2 mm. Size, grade, and lymphvascular invasion of the primary tumor, as well as number, status, size of metastatic disease, and presence of extranodal capsular extension of sentinel and nonsentinel nodes were recorded. Of the 358 cases, 89 had positive sentinel nodes, 29 of which represented micrometastases. Only one (3%) of the 29 cases contained a nonsentinel node with macrometastasis. In 60 of the 89 cases sentinel nodes contained macrometastases. Of these, 38 cases (63%) had metastatic tumor in nonsentinel nodes. Intraoperative consult was performed in 53 of the 89 cases with positive sentinel nodes. Only 1 of the 19 (5%) intraoperative consult cases with micrometastatic sentinel nodes had positive nonsentinel nodes, while 21 of 34 (62%) of macrometastatic sentinel nodes at intraoperative consult had tumor in nonsentinel nodes. No single variable studied discriminated between micro- vs macrometastatic disease. At intraoperative consult, macrometastatic disease was present in all three diagnostic preparations, while diagnostic material in micrometastatic sentinel nodes was usually present in only one modality. This analysis suggests that the risk of finding tumor in nonsentinel nodes differs significantly between cases with micro (3%)- vs macro (63%)-metastatic disease in sentinel nodes. This holds true for cases assessed by intraoperative consult. Considering the known morbidity of complete axillary dissection, assessments of risk vs benefit of undertaking this procedure should be performed on a case-by-case basis in patients with sentinel node micrometastases.

Improving Recognition of Thyroid Carcinoma in Rapid-Consultation Specimens

To improve recognition of thyroid carcinoma in rapid consultation on Diff-Quik-stained (Fisher Diagnostics, Middletown, Virginia, USA.) fine-needle aspiration (FNA) and rapid hematoxylin-eosin (H-E)-stained intraoperative scrape preparation (ISP) specimens by assessing 3 variables (anisokaryosis, nuclear overlap [NO] and scant/absent colloid) in cases of cellular follicular lesions (CFL), an indeterminate diagnostic category.Thirty-seven FNAs and 28 ISPs diagnosed as CFL, with histologic follow-up, were evaluated in blinded fashion by 3 cytopathologists assessing the 3 variables.Over 90% of the malignant cases showed NO in both FNA and ISP, while only 22% of the benign cases did; positive and negative predictive values (PPV and NPV) were 82% and 100%. All malignant cases showed significant anisokaryosis in both FNA and ISP in contrast to 24% of benign cases; PPV and NPV were 74% and 100%. Scant/absent colloid was seen in 87% and 39% of malignancies in FNA and ISP, respectively, as compared to 55% and 20% of the benign cases. PPV and NPV were 52% and 83% in FNA and 63% and 60% in ISP, respectively.Application of these variables improves recognition of thyroid carcinoma, particularly in fine needle aspirates, while additional material may be requested. With ISP, their absence supports recommending against further surgery. Together, optimal surgical planning and outcome may be obtained.

DNA extraction and quantification from touch and scrape preparations obtained from autopsy liver cells

The objective of the present study was to develop a simplified low cost method for the collection and fixation of pediatric autopsy cells and to determine the quantitative and qualitative adequacy of extracted DNA. Touch and scrape preparations of pediatric liver cells were obtained from 15 cadavers at autopsy and fixed in 95% ethanol or 3:1 methanol:acetic acid. Material prepared by each fixation procedure was submitted to DNA extraction with the Wizard genomic DNA purification kit for DNA quantification and five of the preparations were amplified by multiplex PCR (azoospermia factor genes). The amount of DNA extracted varied from 20 to 8,640 microg, with significant differences between fixation methods. Scrape preparation fixed in 95% ethanol provided larger amount of extracted DNA. However, the mean for all groups was higher than the quantity needed for PCR (50 ng) or Southern blot (500 ng). There were no qualitative differences among the different material and fixatives. The same results were also obtained for glass slides stored at room temperature for 6, 12, 18 and 24 months. We conclude that touch and scrape preparations fixed in 95% ethanol are a good source of DNA and present fewer limitations than cell culture, tissue paraffin embedding or freezing that require sterile material, culture medium, laboratory equipment and trained technicians. In addition, they are more practical and less labor intensive and can be obtained and stored for a long time at low cost.

Intraoperative Evaluation of Sentinel Lymph Nodes for Breast Carcinoma: Current Methodologies

Sentinel lymph node biopsy is an important new addition to the surgical management of patients with breast carcinoma. Sentinel nodes have a higher chance of containing metastases than do nonsentinel nodes. Sentinel lymph node biopsy provides an opportunity to stage breast carcinoma patients more accurately and to modify subsequent treatment. One of the most exciting current roles of sentinel lymph node biopsy is the ability to stage patients intraoperatively, allowing a one-step axillary lymph node dissection if the sentinel lymph node contains metastatic carcinoma. Currently, intraoperative evaluation of sentinel lymph nodes is performed using imprint cytology with or without rapid cytokeratin staining, frozen sectioning with or without rapid cytokeratin staining, scrape preparations, or some combination of these techniques. We review the relative strengths and weaknesses of these different methodologies. A great deal of controversy exists regarding the management of patients with metastatic breast carcinoma, particularly those patients with occult and micrometastatic disease. These issues are beyond the scope of this article.

Role of Ultrafast Papanicolaou-Stained Scrape Preparations as an Adjunct to Frozen Sections in the Surgical Management of Thyroid Lesions

To assess the value of intraoperative cytology on scrape preparations of thyroid lesions. We conducted a retrospective review of 73 cases of various thyroid lesions in which ultrafast Papanicolaou-stained scrape preparations were performed. Frozen sections were performed in 19 cases (26%). Fine-needle aspiration diagnosis was available in 65 cases. The correlation between the scrape diagnosis and the final diagnosis was analyzed. The agreement rate between the scrape diagnosis and the final diagnosis was 89%. All cases diagnosed as suspicious for papillary thyroid carcinoma on fine-needle aspiration were definite for malignant papillary carcinoma on scrape preparations. Three cases from this category were diagnosed as papillary carcinoma on both frozen section and scrape preparations, leading to total thyroidectomy. We conclude that intraoperative consultation is useful for thyroid lesions diagnosed as suspicious for papillary thyroid carcinoma on fine-needle aspiration and that ultrafast Papanicolaou-stained scrape preparations can increase the efficacy of thyroid frozen sections and obviate a second procedure for completion thyroidectomy.

Thyroid nodules with FNA cytology suspicious for follicular variant of papillary thyroid carcinoma: follow-up and management.

Thyroid nodules diagnosed as follicular neoplasm on fine-needle aspiration biopsy (FNAB) may represent hyperplastic/adenomatous nodules, follicular adenoma or carcinoma, and follicular variants of papillary thyroid carcinoma (FVPTC) on histologic follow-up. In our laboratory, we attempted to identify a subset of cases which showed cellular specimens with focal features (nuclear chromatin clearing, membrane thickening, and rare grooves) suspicious for the follicular variant of papillary thyroid carcinoma. These cases are reported as follicular-derived neoplasms with nuclear features suspicious for FVPTC to distinguish them from those diagnosed as follicular neoplasm. This study documents our experience with 52 cases so diagnosed and followed prospectively with histologic follow-up. A neoplastic nodule was confirmed in 45/52 cases (86%), of which 40 were malignant (77%). FVPTC was identified in 35/52 cases (67%). Four cases were usual papillary carcinoma, 3 were follicular adenoma, 2 were Hürthle-cell adenoma, and 1 was insular carcinoma. In 7 cases, the subsequent histologic findings were nonneoplastic (5 hyperplastic nodules and 2 colloid nodules). Our prospective study shows that in cellular smears from thyroid nodules, a careful search for the nuclear features of papillary carcinoma should be performed, and it is appropriate to diagnose cases as suspicious for FVPTC if the nuclear features of papillary carcinoma are focal. The surgical management of this group may include an intraoperative confirmation of cytologic diagnosis by scrape preparation and/or frozen section in order to avoid a second surgical intervention for completion thyroidectomy.

Comparison of Aspiration, Touch and Scrape Preparations Simultaneously Obtained from Surgically Excised Specimens

To determine whether three commonly used methods of obtaining and preparing cells for cytologic evaluation--touch preparation (TP), fine needle aspiration (FNA) and scrape preparation--yield essentially comparable material for evaluation or whether important differences exist between them that may influence interpretation. FNA, TP and scrape preparations were made simultaneously on surgically excised specimens. Thirty breast, 6 lung and 12 colon lesions were studied. Each slide was assessed for qualitative and quantitative features. Cellularity was semiquantitatively scored. Scrape preparations were the most cellular (P < .0001). Scrape and TPs often had larger tissue fragments than FNA. However, fragmentation of epithelial groups into smaller clusters and single cells was often noted on scrape preparations. FNAs tended to have cleaner backgrounds than the other two methods. Most often, there are few differences between smears prepared by the three techniques studied. However, scrape preparations may yield smears with smaller clusters and single cells as compared to the other two methods; that is a potential source of diagnostic confusion, particularly with respect to benign breast lesions. Scrape preparations uniformly yield more cellular smears. To the extent that cellularity is an interpretive factor in assessing cytologic specimens, it is important to be aware of the increased cellularity of scrape preparations as compared to the other two techniques.

Immunocytochemistry on cytologic specimens of limited quantity.

Some cytologic specimens may be limited in quantity, and this may hamper or preclude the performance of immunocytochemistry (ICC) in cases where more than one antibody (ab) is required by ICC to arrive at a definitive diagnosis. There is very little information in the cytology literature regarding the use of ICC for specimens that are limited in quantity. In this study, we describe a method, derived from the principles of double immunolabelling, whereby more than one ab test can be repeatedly used on the same Papanicolaou stained slide. Multiple cytologic scrape preparations fixed in 95% ethanol were obtained from fresh surgical specimens including carcinomas of the breast, endometrium, stomach, ovary and colon. Nonneoplastic tissues included tonsil (2), lymph node (2) and myometrium. Papanicolaou stained slides or unstained slides were subjected to two sequential ICC procedures, the first in which the ab was known to be nonreactive with the cells (insulin, glucagon, or somatostatin) and the second in which the ab was known to be positive in the cells. Positive controls for the known positive abs included a single-step ICC procedure as well as the tissue section. The test abs included CAM 5.2, AE1/3, K903, LCA, L26, UCHL-1, s-100, mCEA, GCDFP-15, vimentin, muscle specific actin and desmin. Identical two-step ab procedures were carried out on the tissues from the same surgical specimens. For Papanicolaou stained cytologic specimens, abs were reactive and gave excellent results for the repeat second-step ICC method. There was no false positive or false negative staining. This "repeat ICC" method also gave excellent results on the tissue sections. Immunocytochemistry can be performed more than once on the very same cytologic specimen if the initial ICC antibody attempt is negative. This method may be especially useful in situations where more than one antibody is needed on a very limited cytologic sample size.

Molecular and immunohistochemical analysis of p53 mutations in scrapings and tissue from preinvasive and invasive breast cancer

Mutations of the p53 gene appear to be one of the most common abnormalities in human cancer. Although many studies have been published about p53 alterations in breast cancer, data on molecular biological detection of p53 mutations in in situ lesions are still rare, and the implications for breast cancerogenesis are unclear. Tissue samples from 83 patients with different stages of breast cancer and from 13 patients with benign breast lesions were screened for p53 gene mutations by polymerase chain reaction (PCR) followed by temperature-gradient gel electrophoresis (TGGE). p53 protein accumulation was analysed by immunohistochemistry (IHC). Samples were gained from fresh-frozen tissue, scrapings, or paraffin embedded tissue. Additionally, 23 pairs of primary tumours and corresponding lymph nodes were examined. p53 gene aberrations were found in 55.7% of the infiltrating carcinomas, in 31.5% of the ductal carcinomas in situ (DCIS) and in one atypical ductal hyperplasia. A positive correlation was seen with high-grade tumours and with comedo. There was no statistically significant relationship with respect to age, menopausal status, tumour size, hormone receptor status or lymphatic invasion. Concordance between TGGE and IHC was seen in only 63% of the cases analysed. However, with regard to p53 mutation screening by TGGE, a high significance (P = 0.0008) was seen between standard tissue extraction and our scrape preparation technique. Among 8 pairs of primary tumours and their corresponding lymph node metastases, only 3 harbored identical p53 mutations in the same exon, while in 5 cases with mutant p53 in the primaries, no mutation was seen in the lymph node. Our data indicate that p53 mutations are frequent in breast tumours associated with unfavorable prognosis, including high-grade or the comedo histotype. There is evidence that p53 gene alterations occur early in breast cancerogenesis, as mutations were detected not only in in situ carcinomas but also in atypical ductal hyperplasia.

Experimentelle Untersuchungen über die Regeneration der Aortenendothelzellen: Automatische und visuelle Auswertung von Autoradiogrammen

Autoradiographic studies on plane preparations using 3H-thymidine and pulse cytophotometric investigations on scrape preparations of aortic endothelium from 34 rabbits have brought the following results: By means of multiple 3H-thymidine-injections the number of labelled endothelial cells is higher in young growing animals than in older rabbits; pulse cytophotometrically we did not find any indication of polyploidization processes; from that we conclude a decrease of the growth fraction with advancing age. In comparison to visual counting the automatic image analysis is a highly effective method for the evaluation of autoradiograms. After multiple 3H-thymidine applications the mean blacking intensity of the labelled cell nuclei is lower in 2 months old animals than in 12 months old rabbits; that may mean an increase in the S-phase duration of the proliferating endothelial cells of the older animals. In the area with branching sites there are more 3H-thymidine-labelled endothelial cells than in the neighbouring vascular segments. The significance of these findings for the preferential incidence of arteriosclerosis in this area is discussed.