Histocytological Preparation Techniques ; methods ; Microtomy ; methods ; Tissue Embedding

This study was aimed to establish a smear protocol for preparing bone marrow cells and investigate its effect on fluorescence in situ hybridization (FISH) signal. Probe DNA (C-myc, MDM2, STK6) was labeled with Spectrum Green, PromoFluor-555 and PromoFluor-415 by nick translation. Five bone marrow samples were tested by two methods separately. Traditional method: after removing the erythrocytes by hypoosmotic solution, the bone marrow cells were fixed in methanol/acetic acid (3:1). Improved method: erythrocytes were removed using density gradient centrifugation and fixed in methanol. The samples were then fixed again in 2 formaldehyde for 5 min. The FISH signal was assessed by comparing the relative signal intensity of each fluorophore with the autofluorescence background. The results indicated that improved method greatly increased the ratio of fluorescence signal intensity in the Spectrum Green, PromoFluor-555 and PromoFluor-415 channel (traditional method: 4.3 ± 0.19, 3.52 ± 0.04, 3.07 ± 0.08; improved method: 9.89 ± 0.41, 7.55 ± 0.5, 5.67 ± 0.18, n = 5, P < 0.01) respectively. The signal intensity increased 2.32, 2.14 and 1.85-fold in the Spectrum Green, PromoFluor-555 and PromoFluor-415 channel respectively. In addition, the improved method decreased the split signals [traditional method: (15.8 ± 1.74), (20.42 ± 2.88), (23.2 ± 3.02); improved method: (8.6 ± 1.2), (12.28 ± 1.33), (12.6 ± 2.56), n = 5, P < 0.05]. It is concluded that the improved optimal procedure which facilitates FISH intensity on bone marrow cells is developed, showing potential for wide application in the diagnosis of hematologic diseases.
Bone Marrow Cells ; cytology ; Histocytological Preparation Techniques ; Humans ; In Situ Hybridization, Fluorescence ; methods

Hepatocytes ; pathology ; Histocytological Preparation Techniques ; methods ; Humans ; Liver ; pathology ; Liver Cirrhosis ; pathology

Histocytological Preparation Techniques ; Humans ; Immunohistochemistry ; Microscopy, Electron ; methods ; Molecular Biology ; methods ; Staining and Labeling

OBJECTIVEStudy on the diagnostic accuracy and value of cell block and tissue fragment preparations collected from lung fine needle aspiration (FNA).

METHODSA total of 187 FNA (22G) samples from the lungs with matched histological diagnosis were studied. Among them, the diagnosis made by depending on 124 cell block and fragment preparations were analyzed in comparing retrospectively with the diagnosis of 187 cases by smear preparations.

RESULTS(1) Of the 124 cell blocks cases, 89 cases were true positives, 22 cases were true negatives, 13 cases were false negatives and no false positives. Of the 187 smears cases, the figure were 136, 30, 19 and 2 cases respectively. The diagnostic accuracy of cell blocks was 87.3% in sensitivity, 100% in specificity, 89.5% in overall accuracy. The figures for smears were 87.7%, 93.8% and 88.8% respectively. (2) For malignant tumours, the histological typing accuracy of cell blocks was 93.3% (83/89), and to be 67.9% (91/134) by diagnosis depending on the smears (P < 0.01). For the benign lesions, the figures were 86.4% (19/22) and 60% (18/30) respectively (P < 0.05). (3) It was possible to obtain many minisections for further studies from cell blocks. Immunoperoxidase staining on minisections was reliable and agreed with those on the surgical specimens.

CONCLUSIONSThe diagnostic accuracy of cell block is high, particularly in histological typing which approaches to that of the diagnosis made depending on the postoperative specimens. A combined use of smears and cell block is recommended which may raise further the diagnostic accuracy.

Adolescent ; Adult ; Aged ; Biopsy, Needle ; Female ; Histocytological Preparation Techniques ; Humans ; Lung Neoplasms ; classification ; pathology ; Male ; Middle Aged

Ascitic Fluid ; cytology ; Cytological Techniques ; instrumentation ; methods ; Equipment Design ; Histocytological Preparation Techniques ; instrumentation ; methods ; Humans ; Paraffin Embedding ; Specimen Handling ; instrumentation ; methods

The task of segmenting histiocyte is a crucial step in the analysis of histiocytic images which is an important application of computer vision to histopathology. The algorithm presented in this article was composed of two steps: (1) the morph-based preprocessing; (2) the ameliorated watershed method. In the first step, the difference between histiocytes was magnified in order to increase the visibility from the view of the computer vision, and then the ameliorated terrace-flooding-simulated watershed method was used to achieve the segmentation of histiocytic images in the second step. To test the performance of the algorithm, different samples of visual quality were tested and the result figures proved successful.
Algorithms ; Bone Marrow Cells ; cytology ; Cartilage ; cytology ; Histocytological Preparation Techniques ; methods ; Humans ; Image Processing, Computer-Assisted ; methods ; Numerical Analysis, Computer-Assisted ; Pattern Recognition, Automated ; methods

Breast Neoplasms ; pathology ; surgery ; Carcinoma, Ductal, Breast ; pathology ; surgery ; Cryoultramicrotomy ; Cytodiagnosis ; methods ; Female ; Histocytological Preparation Techniques ; methods ; Humans ; Intraoperative Care ; methods ; Precancerous Conditions ; pathology ; surgery

Objective. To study the clinical spectrum of Filipino patients with Williams Syndrome and to confirm the gene deletion by FISH analysis. Methods. From June 2005 to September 2008, patients who were seen at the Genetics clinic of the UP-PGH and who met the clinical criteria for Williams Syndrome were analyzed for the 7q11.23 deletion through karyotyping and FISH studies. A detailed history and a thorough dysmorphologic examination were performed. Relevant investigations included two-dimensional echocardiography, renal ultrasonography, ophthalmologic examination, developmental assessment and serum calcium determination. Result. Eight patients were included in the study. The mean age at first diagnosis was 8.5 years. All cases were sporadic. The chromosomal analysis was normal for all patients and in the FISH analysis, a 7q11.23 deletion was detected in 100% of cases. Distinctive facial features, cardiac abnormalities and developmental delay were present in all patients. The typical behavior of overfriendliness was observed in the majority of cases. Hypercalcemia was documented in only one case and no renal anomalies were detected. Conclusion. The craniofacial features were similar among patients but there is a broad spectrum of severity of clinical features in cardiovascular abnormalities, personality, behavior traits and mental capacity.
CYTOGENETICS ; GENETICS ; WILLIAMS SYNDROME ; NERVOUS SYSTEM DISEASES ; NEUROLOGIC MANIFESTATIONS ; NEUROBEHAVIORAL MANIFESTATIONS ; INTELLECTUAL DISABILITY ; GENE DELETION ; IN SITU HYBRIDIZATION, FLUORESCENCE ; AORTIC STENOSIS, SUPRAVALVULAR ; DIAGNOSIS ; DIAGNOSTIC TECHNIQUES AND PROCEDURES ; CLINICAL LABORATORY TECHNIQUES ; CYTOLOGICAL TECHNIQUES ; HISTOCYTOLOGICAL PREPARATION TECHNIQUES ; STAINING AND LABELING ; IN SITU HYBRIDIZATION ; ;

OBJECTIVEThe purpose of this study was to explore the value of thin-layer cytology (TLC) in intraoperative fine needle aspiration cytology diagnosis of pancreatic cancer.

METHODSResults of cytological examination with thin-layer smears were compared with that with conventional smears in intraoperative fine-needle aspiration (FNA) biopsy.

RESULTSTotally 271 fine needle aspiration biopsies were performed, among them, 70 were examined with thin-layer smears, showing unsatisfactory smear in 5 cases (7.1%); 201 were examined with conventional smears (CS), showing unsatisfactory smear in 9 cases (4.5%). No significant difference in the unsatisfactory smears was observed between those two groups. The positive rate of diagnosis with CS smears was 60.0% (42/70) and that of TLC was 81.6% (164/201), with a significant difference (P < 0.01). The sensitivity of CS and TLC was 68.9% and 87.7%, respectively (P < 0.01). The sensitivity of both FNA and frozen section diagnosis in 20 cases was 90.0%, respectively, but that of FNA combined with frozen section diagnosis was 95.0%. 9 cancer cases diagnosed by pathology were initially negative by cytology, but adenocarcinoma cells were found in 7 cases of them by the second time cytology examination.

CONCLUSIONThe positive rate is high in TLC smears, and unsatisfactory rate is low. TLC smears are one of the best methods in intraoperative confirmation of pancreatic cancer. The use of FNA smears combined with frozen section biopsy can further improve the sensitivity of diagnosis.

Adult ; Aged ; Aged, 80 and over ; Biopsy, Fine-Needle ; methods ; Cytodiagnosis ; Female ; Frozen Sections ; Histocytological Preparation Techniques ; methods ; Humans ; Intraoperative Period ; Male ; Middle Aged ; Pancreatic Neoplasms ; diagnosis ; pathology ; Young Adult