Frozen tissue preparation for high-resolution multiplex histological analyses of human brain specimens
Fangjie ShaoWenhong JiangQingqing GaoBaizhou LiChongran SunQiyuan WangQin ChenBing SunHong ShenKeqing ZhuJianmin ZhangChong Liu
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Brain tissue
Multiplex
Objective:To discuss the clinical application of frozen section in thyroid operation so as to raise the rate of correct diagnosis. Methods: A retrospective review was made for 356 frozen sections and paraffin sections. Results: The frozen section diagnosis showed out 309 cases with benign lesion , 36 cases with malignant tumor and there were 11delayed diagnosis ;in the control , the paraffin section showed 314 cases with benign lesion and 42 cases with malignant tumor ; There were 5 false-negative diagnosis dy frozen section (1.4%) and conformed dinagnosis rate of frozen section reached 98.5%. Conclusion: Correct diagnosing rate of frozen section in thyroid may be raised by increasing the section quality and improving the method.
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Frozen section is known to be a valuable tool in the setting of indistinct lesions, lesions in cosmetically or functionally important areas, and those with recurrent or residual tumors. Most non–Mohs surgery studies comparing frozen section with paraffin sections suggest a concordance rate of 85% to 90%, whereas studies with Mohs surgery often suggest concordance rates of 95% to 98%. We do not perform Mohs surgery at our institutions but feel that the accuracy of frozen section is relatively high. Frozen-section data from between 2005 and 2011 was analyzed, and a total of 150 cases was found and assessed. Most of the cases were basal cell carcinomas and squamous cell carcinomas, with most arising in the head and neck region. Half of the resections were for previous incomplete margins with the other half being primary excisions. The frozen section was accurate in 97.7% of the cases when compared with the formal paraffin sections. However, the incomplete rate was higher at 14.8% because of patients with known positive margins on frozen section and the use of less accurate techniques of specimen analysis. We feel that, when used appropriately, frozen section can be a reliable tool and that a negative result should provide the surgeon enough reassurance as to undertake immediate reconstruction.
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Mohs surgery
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Objective:To probe a practical and convenient approach of reducing the detachment of brain tissue sections from glass slides and improving the of experiments.Methods:The new methods was studied in the progresses of treating glass slides,brain tissue perfusion fixation,frozen section and immunohistochemistry.Results:Treating of chromic potassium sulfate on glass slides reduced the detachment of brain tissue section;cold NS and 4% paraformaldehyde improved tissue perfusion fixation;the method,that liquid nitrogen frozen brain tissue quickly,reduced ice crystal.Conclusion:The detachment of brain sections can be reduced;immunohistochemical results of brain frozen section can be improved,the method is worth spreading among clinical and scientific researches.
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Brain tissue
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Head and neck surgeons commonly request frozen sections. Practice patterns vary from laboratory to laboratory on how the tissue is used in performing the frozen section. Some pathologists wish to see all the material submitted by consuming it completely during frozen section, whereas others reserve some for permanent section. We wished to determine whether knowledge of margin status was initially inaccurate because of reserving tissue for permanent section.Sixty-five laryngectomies (total and partial) with margin assessment enhanced by frozen section evaluation were studied. Forty-five laryngectomy specimens, generating 249 frozen sections in which a permanent section was prepared from tissue remaining from frozen section examination, were studied.Five of the 249 frozen sections contained a discrepancy between the frozen section and permanent section because of insufficient leveling of the frozen section block. These five discrepancies were called negative on frozen section, but permanent section revealed dysplasia (two cases of mild dysplasia, one case with moderate dysplasia, and one case with severe dysplasia) or carcinoma in situ (one case). Twenty laryngectomies in which the frozen section tissue was consumed at the time of frozen section generated 103 frozen sections. In eight of the frozen sections involving six cases, the diagnostic tissue was not present on one or two of the frozen section levels examined.We conclude that in examining margins for laryngeal squamous cell carcinoma the frozen section tissue should be completely sampled by examining several levels at the time of frozen section. This requires consuming or exhausting the frozen section tissue rather than reserving any remaining frozen tissue for a paraffin-embedded permanent section.
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Objective To compare the effects of low temperature fixation ordinary temperature fixation on freezing tissue. Methods The remaining anolonely tissue (colon and stomach) after frozen section of 48 SD rats were randomly divided into 2 groups:low temperature fixation (LTF) and ordinary temperature fixation (OTF). LTF was treated by 4 ℃4% paraformaldehyde for 1 hour,fixated in room temperature for 12 hours,dehydration and paraffin imbedding by routine method. OTF was fixation in room temperature,dehydration and paraffin imbedding by routine method. Both groups were stained by hematoxylin-eosin staining,oluidine blue staining and immunohistochemical staining methods respectively,being observed by microscope. Results It was shown that the cellular stucture was clearer in LTF than that in OTF. The colour of the paraffin section was bright in LTF. The contrast of the nuclear and cytoplasm in LTF was clear. We could see more spacing caused by ice crystal in paraffin section of OTF. Conclusion There is no obviously differences in immunohistochemical slices.The study indicates that the effect of low temperature fixation of the freezing tissue of paraffin section is better than that of ordinary temperature fixation.
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Eosin
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The study evaluates the morphology of frozen tissue sections treated by microwave-stimulated fixation compared with unfixed cryostat sections. Furthermore, microwave fixation has been compared with fixation at room temperature. Improved preservation of nuclear structures (chromatin pattern and nucleoli) was found with the microwave combination, while other parameters were indistinguishable from the unfixed cryostat sections. Fixation at room temperature showed no difference compared with the unfixed cryostat sections. The combined method might be useful in rapid diagnosis of mesenchymal tumors, lymphoid proliferations and tumors of the CNS.
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Objective To discuss the clinical application of frozen section in thyroid operation so as to raise the rate of correct diagnosis.Methods A retrospective review was made to 248 frozen sections and paraffin sections.Results The frozen section diagnosis showed out 217 cases with benign lesion,22 cases with malignant tumor and there were 9 delayed diagnosis;in the control,the paraffin section showed 220 cases with benign lesion and 28 cases with malignant tumor;There were 3 false-negative diagnosis by frozen section (1.21%) and conformed diagnosis rate of frozen section reached 95.1%.Conclusions The correct diagnosing rate of frozen section in thyroid operation may be raised by increasing the section quality and improving the method.
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Post-mortem brain MRI can yield valuable information. However, tissue preservation requires substitution of the CSF-fluid by fixation agents, which is time-consuming for large samples. R 2 * maps, dominated by R 2 -effects, were measured at several timepoints during fixation of pig-brain tissue samples using formalin-based fixatives.
Brain tissue
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Fixation time
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Objective:To evaluate the diagnostic accuracy of intraoperative frozen section in breast lesions.Methods:To analyze frozen section and paraffin section diagnosis of 555 cases of breast lesions.Results:Total of 546 cases(98.38%) of intraoperative frozen sections were diagnosed correctly and 7 cases were awaiting definitive diagnosis and 2(0.59%) was false negative diagnosis.Conclusion:Intraoperative frozen section diagnosis of breast lesions is an important means of diagnosis.
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