scholarly journals Rapid brain structure and tumour margin detection on whole frozen tissue sections by fast multiphotometric mid-infrared scanning

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tim Kümmel ◽  
Björn van Marwick ◽  
Miriam Rittel ◽  
Carina Ramallo Guevara ◽  
Felix Wühler ◽  
...  
Keyword(s):  
Imaging System ◽  
Frozen Section ◽  
Computational Effort ◽  
Primary Hepatocellular Carcinoma ◽  
Measuring System ◽  
Sufficient Information ◽  
Tissue Samples ◽  
Mid Infrared ◽  
Tissue Sections ◽  
Frozen Tissue

AbstractFrozen section analysis is a frequently used method for examination of tissue samples, especially for tumour detection. In the majority of cases, the aim is to identify characteristic tissue morphologies or tumour margins. Depending on the type of tissue, a high number of misdiagnoses are associated with this process. In this work, a fast spectroscopic measurement device and workflow was developed that significantly improves the speed of whole frozen tissue section analyses and provides sufficient information to visualize tissue structures and tumour margins, dependent on their lipid and protein molecular vibrations. That optical and non-destructive method is based on selected wavenumbers in the mid-infrared (MIR) range. We present a measuring system that substantially outperforms a commercially available Fourier Transform Infrared (FT-IR) Imaging system, since it enables acquisition of reduced spectral information at a scan field of 1 cm2 in 3 s, with a spatial resolution of 20 µm. This allows fast visualization of segmented structure areas with little computational effort. For the first time, this multiphotometric MIR system is applied to biomedical tissue sections. We are referencing our novel MIR scanner on cryopreserved murine sagittal and coronal brain sections, especially focusing on the hippocampus, and show its usability for rapid identification of primary hepatocellular carcinoma (HCC) in mouse liver.

scholarly journals Molecular imaging can identify the location to perform a frozen biopsy during intraoperative frozen section consultation

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0252731
Author(s):  
Mitchell G. Bryski ◽  
Lydia G. Frenzel-Sulyok ◽  
E. James Delikatny ◽  
Charuhas Deshpande ◽  
Leslie A. Litzky ◽  
...  
Keyword(s):  
Molecular Imaging ◽  
Murine Model ◽  
Sampling Error ◽  
Frozen Section ◽  
Fluorescent Microscopy ◽  
Surgical Oncology ◽  
Tissue Samples ◽  
Surgical Specimen ◽  
Intraoperative Frozen Section ◽  
Tissue Sections

Background Intraoperative frozen section (FS) consultation is an important tool in surgical oncology that suffers from sampling error because the pathologist does not always know where to perform a biopsy of the surgical specimen. Intraoperative molecular imaging is a technology used in the OR to visualize lesions during surgery. We hypothesized that molecular imaging can address this pathology challenge in FS by visualizing the cancer cells in the specimen in the pathology suite. Here, we report the development and validation of a molecular-imaging capable cryostat called Smart-Cut. Methods A molecular imaging capable cryostat prototype was developed and tested using a murine model. Tumors grown in mice were targeted with a NIR contrast agent, indocyanine green (ICG), via tail vein injection. Tumors and adjacent normal tissue samples were frozen sectioned with Smart-Cut. Fluorescent sections and non-fluorescent sections were prepared for H&E and fluorescent microscopy. Fluorescent signal was quantified by tumor-to-background ratio (TBR). NIR fluorescence was tested in one patient enrolled in a clinical trial. Results The Smart-Cut prototype has a small footprint and fits well in the pathology suite. Fluorescence imaging with Smart-Cut identified cancerous tissue in the specimen in all 12 mice. No false positives or false negatives were seen, as confirmed by H&E. The mean TBR in Smart-Cut positive tissue sections was 6.8 (SD±3.8). In a clinical application in the pathology suite, NIR imaging identified two lesions in a pulmonary resection specimen, where traditional grossing only identified one. Conclusion Molecular imaging can be integrated into the pathology suite via the Smart-Cut device, and can detect cancer in frozen tissue sections using molecular imaging in a murine model.

Mucin Stain on Frozen Sections

1999 ◽  
Vol 123 (5) ◽  
pp. 378-380
Author(s):  
Suchetha Soans ◽  
Lorenzo M. Galindo ◽  
Fernando U. Garcia
Keyword(s):  
Colonic Mucosa ◽  
Frozen Section ◽  
Staining Technique ◽  
Turnaround Time ◽  
Staining Procedure ◽  
Normal Colonic Mucosa ◽  
Frozen Sections ◽  
Tissue Sections ◽  
Paget Disease ◽  
Frozen Tissue

Abstract Context.—The importance of frozen-section diagnoses in the practice of pathology cannot be overemphasized. In some cases, the use of a mucin stain can greatly aid in the diagnosis. Since few methods for mucin staining have been described that can be used in the frozen-section setting, we developed one such staining procedure for mucin. Objective.—To develop a rapid mucicarmine staining technique to be used on frozen sections that does not significantly delay overall turnaround time. Design.—A standard mucicarmine staining technique was modified by using a concentrated mucicarmine stain and a microwave oven, to reduce the total staining time to 3 minutes or less. Frozen tissue from normal colonic mucosa was used as a control, and skin from extramammary Paget disease for evaluation of margins was used as a case. Results.—The rapid mucicarmine stain successfully demonstrated the presence of mucin on frozen-tissue sections. Mucin stained deep rose, and the connective tissue stained green. Conclusion.—This rapid and simple mucin staining technique can be used on frozen sections with no significant effect on the overall turnaround time, thereby aiding rapid diagnosis on frozen sections.

scholarly journals Role of Intraoperative Frozen Section Diagnosis in the Management of Patients with CNS Lesions: Evaluation of 60 Consecutive Intraoperative Frozen Sections from 1991 to 1993

2020 ◽  
Vol 1 (1) ◽  
Author(s):  
Seyed Mirsattari ◽  
Fraser W. Saunders
Keyword(s):  
Frozen Section ◽  
Imaging Techniques ◽  
Frozen Sections ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Intraoperative Management ◽  
Clinical Diagnoses ◽  
Cns Lesions

To determine the role of intraoperative frozen sections (FSs) in the management of patients with central nervous system (CNS) lesions, 60 consecutive intraoperative clinical diagnoses of CNS lesions were presented and compared with concomitantly obtained FS diagnoses. Clinical diagnoses were established byhistory, physical examination, imaging techniques, and gross appearance of the abnormal tissue in situ. Tissue samples were obtained intraoperatively and processed for FS diagnoses. The findings of the FS diagnoses were reported to the operating room and compared with the clinical diagnoses. The remainingbiopsy samples were used to prepare paraffin-embedded tissue sections from which the definitive diagnoses were made. Comparison of the clinical and FS diagnoses, using paraffin-embedded tissue as the true diagnosis, shows that FS diagnosis has a limited contribution to intraoperative patient management by the neurosurgeon. The rate of diagnostic failures between the two techniques was very similar; clinical diagnoses and FSs were misinterpreted in 12 and 11 of the 60 cases, respectively. Compared to a clinical diagnosis, the intraoperative FS technique provided no significant improvement in diagnosis and management; it altered the intraoperative management of the patients in 2 of 60 cases.

Discovery of intracellular 2,8 dihydroxyadenine crystals in bovine lymphatic and hepatic cells

1987 ◽  
Vol 45 ◽  
pp. 906-907
Author(s):  
W. E. Rigsby ◽  
D. M. Hinton ◽  
V. J. Hurst ◽  
P. C. McCaskey
Keyword(s):  
Polarized Light ◽  
Paraffin Sections ◽  
Tissue Samples ◽  
Whole Cells ◽  
Tissue Sections ◽  
Hepatic Cells ◽  
Slaughter House ◽  
Mammalian Tissues ◽  
Carbon Coated ◽  
Cellular Components

Crystalline intracellular inclusions are rarely seen in mammalian tissues and are often difficult to positively identify. Lymph node and liver tissue samples were obtained from two cows which had been rejected at the slaughter house due to the abnormal appearance of these organs in the animals. The samples were fixed in formaldehyde and some of the fixed material was embedded in paraffin. Examination of the paraffin sections with polarized light microscopy revealed the presence of numerous crystals in both hepatic and lymph tissue sections. Tissue sections were then deparaffinized in xylene, mounted, carbon coated, and examined in a Phillips 505T SEM equipped with a Tracor Northern X-ray Energy Dispersive Spectroscopy (EDS) system. Crystals were obscured by cellular components and membranes so that EDS spectra were only obtainable from whole cells. Tissue samples which had been fixed but not paraffin-embedded were dehydrated, embedded in Spurrs plastic, and sectioned.

Raman molecular imaging of brain frozen tissue sections

2014 ◽  
Vol 120 (1) ◽  
pp. 55-62 ◽  
Author(s):  
Rachel E. Kast ◽  
Gregory W. Auner ◽  
Mark L. Rosenblum ◽  
Tom Mikkelsen ◽  
Sally M. Yurgelevic ◽  
...  
Keyword(s):  
Molecular Imaging ◽  
Tissue Sections ◽  
Frozen Tissue

Surface immunoglobulin restriction in B-non Hodgkin's lymphomas in cell suspension and on frozen tissue sections

2009 ◽  
Vol 35 (4) ◽  
pp. 399-407 ◽  
Author(s):  
Philip M. Kluin ◽  
Roel A. Weger ◽  
Henk-Jan Schuurman ◽  
Peter P. J. Peters ◽  
Philomé I. Spies ◽  
...  
Keyword(s):  
Cell Suspension ◽  
Tissue Sections ◽  
Surface Immunoglobulin ◽  
Frozen Tissue ◽  
Hodgkin's Lymphomas

Frozen Section Examination of Liver, Gallbladder, and Pancreas

2005 ◽  
Vol 129 (12) ◽  
pp. 1610-1618 ◽  
Author(s):  
Juan Lechago
Keyword(s):  
Frozen Section ◽  
Gallbladder Wall ◽  
Confounding Factors ◽  
Tertiary Referral ◽  
Pertinent Literature ◽  
Medical Centers ◽  
Frozen Tissue ◽  
Relative Rarity ◽  
Gross Examination ◽  
Frequent Presence

Abstract Context.—Frozen section of the liver is a comparatively frequent request that most often applies to a relatively limited number of situations. The only indication for frozen tissue examination of a gallbladder with any frequency is the presence of a polypoid mucosal lesion or a suspicious thickening of the gallbladder wall. A variety of intraoperative consultations may be applicable during surgery of the pancreas. Objective.—To examine the indications and pitfalls regarding the gross examination and frozen section performance for liver, gallbladder, and pancreas. Data Sources.—Author experience and review of the pertinent literature. Conclusions.—Although indications are relatively straightforward for frozen section of liver and gallbladder, handling of the pancreas specimens for frozen tissue examination is often a cause for a certain degree of anxiety. This situation is the result of a relative rarity of such specimens outside large tertiary referral medical centers coupled with a variety of confounding factors, including the presence of chronic pancreatitis with distortion of the normal structures and the frequent presence of variable degrees of dysplasia. The suboptimal preservation of the frozen tissue adds further angst to the scenario. In this article, the main issues are critically examined in light of the experience of the author and others.

Visualisation of GABAergic neurons and synapses in the rat brain using immunohistochemistry for two forms of glutamate decarboxylase

2021 ◽  
Vol 21 (2) ◽  
pp. 63-73
Author(s):  
Valeria A. Razenkova ◽  
Dmitrii E. Korzhevskii
Keyword(s):  
Brain Tissue ◽  
Glutamate Decarboxylase ◽  
Brain Structures ◽  
Gabaergic Neurons ◽  
Laboratory Animals ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Background Staining ◽  
The Central Nervous System ◽  
Rat Forebrain

BACKGROUND: Taking into account the importance of GABAergic brain system research and also the opportunity to achieve specific and accurate results in laboratory studies using immunohistochemical approaches, it seems important to have a reliable method of visualization GABA-synthesizing cells, their projections and synapses, for the morphofunctional analysis of GABAergic system both in normal conditions and in the experimental pathology. AIM: The aim of the study was to visualize analyze GABAergic neurons and synapses within rats brain using three different antibody types against glutamate decarboxylase and to identify the optimal conditions for reaction performing. MATERIALS AND METHODS: The study was performed on paraffin brain tissue sections of 5 adult Wistar rats. Immunohistochemical reactions using three antibody types against glutamate decarboxylase isoform 67 (GAD67) and glutamate decarboxylase isoform 65 (GAD65) were performed. Additional controls on C57/Bl6 mice and Chinchilla rabbits brain samples were also carried out. RESULTS: Antibodies used in the research made it possible to achieve high quality of GABAergic structures visualizing without increasing background staining. At the same time different antibody types are distinct in their efficacy to perform immunohistochemistry reaction on laboratory animal brain tissue samples. By performing additional controls, we discovered that there is necessary to adsorb secondary reagents immunoglobulins in order to eliminate nonspecific staining. It was found that GAD67 and GAD65 distribution in rat forebrain structures is different. It was stated that GAD67 immunohistochemistry most completely reveals GABAergic brain structures compared to GAD65 immunhistochemistry. The possibility of determining morphological features of GABAergic neurons and synaptic terminals, as well as performing quantitative analysis, was demonstrated. CONCLUSIONS: The approach proposed makes it possible to specifically visualize GABAergic structures of the central nervous system of different laboratory animals. This could be useful both in fundamental studies and in pathology research.

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