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- 摘要
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- 实验方案
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The potential of biobanked liquid based cytology samples for cervical cancer screening using Raman spectroscopy.
摘要： Patient samples are unique and often irreplaceable. This allows biobanks to be a valuable source of material. The aim of this study was to assess the ability of Raman spectroscopy to screen for histologically confirmed cases of Cervical Intraepithelial neoplasia (CIN) using biobanked liquid based cytology (LBC) samples. Two temperatures for long term storage were assessed; 80oC and -25oC. The utility of Raman spectroscopy for the detection of CIN was compared for fresh LBC samples and biobanked LBC samples. Two groups of samples were used for the study with one group associated with disease (CIN3) and the other associated with no disease (cytology negative). The data indicates that samples stored at -80oC are not suitable for assessment by Raman spectroscopy due to a lack of cellular material and the presence of cellular debris. However, the technology can be applied to fresh LBC samples and those stored at -25oC and is, moreover, effective in the discrimination of negative samples from those where CIN 3 has been confirmed. Pooled fresh and biobanked samples are also amenable to the technology and achieve a similar sensitivity and specificity for CIN 3. This study demonstrates that cervical cytology samples stored within biobanks at temperatures that preclude cell lysis can act as a useful resource for Raman spectroscopy and will facilitate research and translational studies in this area.
关键词： cervical pre-cancer,cytology,ThinPrep,Raman spectroscopy,cervical cancer,biobank
更新于2025-09-23 15:22:29
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Fluorescence in situ hybridization in 1?mL of selective urine for the detection of upper tract urothelial carcinoma: a feasibility study
摘要： Kidney-sparing surgery of upper tract urothelial carcinoma (UTUC) requires a stringent follow-up with frequent ureteroscopies. Triage testing could reduce the number of follow-up ureteroscopies and hence minimize the invasiveness of follow-up. The use of urine-based markers for triage seems appealing but should be feasible with selective urine from outpatient cystoscopy to maximize the reduction of invasiveness. In this study, the feasibility of UroVysion? fluorescence in?situ hybridization (FISH) for the detection of UTUC in 1?mL of selective urine is investigated. Ten consecutive patients with biopsy-proven UTUC and five patients with negative diagnostic ureteroscopy findings were included in this case-control study. During ureteroscopy, 1?mL of selective urine was collected passively with a ureteral splint for Urovysion? FISH. The FISH rater was blinded to any clinical information. The results of FISH were compared to the findings of concomitantly collected selective urine cytology and the patients’ UTUC status. FISH was feasible in all samples with a sensitivity of 90% and a specificity of 80% for UTUC. In comparison, selective cytology resulted in a diagnostic yield of 87% with a sensitivity of 80% and a specificity of 67%. In conclusion,?UTUC detection is feasible with FISH in 1?mL of passively collected selective urine. Thus from a technical point of view, FISH could be used as an outpatient triage test to decide if follow-up ureteroscopy is necessary after kidney-sparing surgery of UTUC. Evaluation of the diagnostic accuracy of FISH for the suggested pathway deserves further attention.
关键词： Ureteroscopy,Urothelial carcinoma,Cystoscopy,Fluorescence in?situ hybridization,Urine cytology,Upper tract urothelial carcinoma
更新于2025-09-23 15:21:21
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Raman spectroscopic detection of high-grade cervical cytology: Using morphologically normal appearing cells
摘要： This study aims to detect high grade squamous intraepithelial cells (HSIL) by investigating HSIL associated biochemical changes in morphologically normal appearing intermediate and superficial cells using Raman spectroscopy. Raman spectra (n = 755) were measured from intermediate and superficial cells from negative cytology ThinPrep specimens (n = 18) and from morphologically normal appearing intermediate and superficial cells from HSIL cytology ThinPrep specimens (n = 17). The Raman data was subjected to multivariate algorithms including the standard principal component analysis (PCA)-linear discriminant analysis (LDA) and partial least squares discriminant analysis (PLS-DA) together with random subsets cross-validation for discriminating negative cytology from HSIL. The PCA-LDA method yielded sensitivities of 74.9%, 72.8%, and 75.6% and specificities of 89.9%, 81.9%, and 84.5%, for HSIL diagnosis based on the dataset obtained from intermediate, superficial and mixed intermediate/superficial cells, respectively. The PLS-DA method provided improved sensitivities of 95.5%, 95.2% and 96.1% and specificities of 92.7%, 94.7% and 93.5% compared to the PCA-LDA method. The results demonstrate that the biochemical signatures of morphologically normal appearing cells can be used to discriminate between negative and HSIL cytology. In addition, it was found that mixed intermediate and superficial cells could be used for HSIL diagnosis as the biochemical differences between negative and HSIL cytology were greater than the biochemical differences between intermediate and superficial cell types.
关键词： cervical cytology,Raman spectroscopy,multivariate algorithms,biochemical changes,HSIL
更新于2025-09-23 15:21:21
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Sensitive and Rapid Cancer Diagnosis with Immunoplasmonic Assay Based on Plasmonic Nanoparticles: Toward Fine-Needle Aspiration Cytology
摘要： Fine-needle aspiration cytology is a minimally invasive diagnostic strategy. However, collected specimens have to undergo a series of complex preparation steps, thus limiting the use of fine-needle aspiration cytology as a diagnostic approach for cytopathologists. In this study, we propose a cytology approach, easily adaptable to conventional one, by employing plasmonic biomarkers (gold(red_scattering), silver/gold(blue_scattering), and silver/gold(green_scattering) nanoparticles), each having a well-defined optical feature. Plasmonic nanoparticles, incubated with the suspension cells, are fully visible and differentiable on the cell membranes under the darkfield lateral RGB (Red-Green-Blue) side-illumination microscopy. The post-fabrication of plasmonic nanoparticles by the conjugation of a specific antibody (anti-human epidermal growth factor receptor 2: HER2) directly allows to visualize the selectively recognized antigens of the suspension cells (MDA-MB-453++HER2 and Jurkat-HER2) in co-cultures and individual cultures by using the side-illumination compared to the conventional dark-field visualization protocol. We also incubate two cancer cell lines (trypsinized MDA-MB-231++CD44 and MDA-MB-453++HER2) with anti-CD44 and anti-HER2 functionalized plasmonic nanoparticles, and nano-immunoplasmonic and immunofluorescence assays show a good agreement with each other for the recognition of expressed antigens (CD44 and HER2). Our results demonstrate a remarkable potential of plasmonic nanoparticles for a simple and rapid examination of small population of cells from collected specimens by using nano-immunoplasmonic approach and the side-illumination microscopy.
关键词： Fine-needle aspiration cytology,Plasmonic nanoparticles,Plasmonic biomarkers,Cancer diagnosis,Immunoplasmonic Assay,Side-illumination microscopy
更新于2025-09-23 15:19:57
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Synovial Sarcoma Diagnosis on Fine Needle Cytology Sample Confirmed by Fluorescence in Situ Hybridization
摘要： Synovial sarcoma (SS) is a mesenchymal neoplasm usually affecting adolescents and young adults, with one third of cases occuring in the first two decades of life. SS is characterized by the specific balanced chromosomal translocation t(X;18)(p11;q11), resulting in the fusion of SYT gene on chromosome 18 with SSX1, SSX2 or SSX4 gene on chromosome X. SS predominantly arises in the soft tissue around large joints of the lower extremities. Other sites include head and neck, thorax, heart or pericardium, abdomen and pelvis. Exceedingly rare cases have been reported in an intramuscular location at the shoulder. The cytological diagnosis of a monophasic synovial sarcoma can be challenging, due to the wide spectrum of differential diagnosis. Here we describe a rare case of an intramuscular monophasic synovial sarcoma diagnosed by an integrated cytological and molecular approach.
关键词： Synovial sarcoma,Intramuscular,Fine needle cytology,Monophasic synovial sarcoma,Fluorescence in situ hybridization
更新于2025-09-10 09:29:36
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Molecular Diagnostics in Cytopathology (A Practical Handbook for the Practicing Pathologist) || Fluorescence In Situ Hybridization (FISH) Testing in Urinary Tract Cytology
摘要： Fluorescence in situ hybridization (FISH) allows for visualization of specific DNA sequences and can, therefore, be used for quantitation of chromosomes and genes, including aneusomies, chromosomal deletions, or amplifications. The commercial assay UroVysion? (Vysis, Inc., Downers Grove, IL, USA) has made the FISH technique available for routine use in hospital cytology laboratories, but most UroVysion? tests are performed at large reference laboratories. The UroVysion? assay is composed of four single-stranded fluorescently labeled nucleic acid probes – three chromosome enumeration probes (CEP) for the chromosomes 3, 7, and 17 and the single locus-specific identifier (LSI) probe 9p21. The DNA probes are directly labeled with the four different fluorescent dyes SpectrumRed (CEP3), SpectrumGreen (CEP7), SpectrumAqua (CEP17), and SpectrumGold (LSI 9p21).
关键词： UroVysion?,locus-specific identifier,Fluorescence in situ hybridization,urothelial carcinoma,urinary cytology,FISH,chromosome enumeration probes
更新于2025-09-10 09:29:36
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Vibrational Microspectroscopy for Cancer Screening
摘要： Vibrational spectroscopy analyses vibrations within a molecule and can be used to characterise a molecular structure. Raman spectroscopy is one of the vibrational spectroscopic techniques, in which incident radiation is used to induce vibrations in the molecules of a sample, and the scattered radiation may be used to characterise the sample in a rapid and non-destructive manner. Infrared (IR) spectroscopy is a complementary vibrational spectroscopic technique based on the absorption of IR radiation by the sample. Molecules absorb specific frequencies of the incident light which are characteristic of their structure. IR and Raman spectroscopy are sensitive to subtle biochemical changes occurring at the molecular level allowing spectral variations corresponding to disease onset to be detected. Over the past 15 years, there have been numerous reports demonstrating the potential of IR and Raman spectroscopy together with multivariate statistical analysis techniques for the detection of a variety of cancers including, breast, lung, brain, colon, oral, oesophageal, prostate and cervical cancer. This paper discusses the recent advances and the future perspectives in relation to cancer screening applications, focussing on cervical and oral cancer.
关键词： cytology,oral cancer,Raman spectroscopy,FTIR spectroscopy,optical diagnosis,cervical cancer,lasers,screening
更新于2025-09-04 15:30:14
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A spontaneously resolving breast lesion: imaging and cytological findings of nodular fasciitis of the breast with FISH showing USP6 gene rearrangement
摘要： We report a case of nodular fasciitis of the breast in a 48-year-old woman who presented with a tender rapidly growing right breast lump. Ultrasound guided fine needle aspiration (FNA) of the solid mass was performed. Cytology was reported as atypical spindle cell neoplasm and the patient was referred to a breast surgeon at a tertiary institution for a definitive diagnosis and further management. Follow-up ultrasound showed partial regression and MRI, mammogram after 2–3 weeks confirmed spontaneous and total resolution of the lesion. Nodular fasciitis of the breast is rarely diagnosed on cytology alone and a histological diagnosis is usually required for a definitive diagnosis. However, in this case, the lesion spontaneously resolved prior to core biopsy or diagnostic open biopsy. The cytological features in conjunction with immunohistochemistry and the clinical history strongly suggest nodular fasciitis, which is further supported by a USP6 FISH positive result.
关键词： cytology,USP6 gene rearrangement,nodular fasciitis,breast,spontaneous resolution
更新于2025-09-04 15:30:14
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Fluorescence imaging of invasive head and neck carcinoma cells with integrin αvβ6-targeting RGD-peptides: an approach to a fluorescence-assisted intraoperative cytological assessment of bony resection margins
摘要： We assessed the use of peptides containing arginylglycylaspartic acid (RGD) that target integrin (cid:2)v(cid:3)6 as a potential approach for a fluorescence-assisted intraoperative cytological assessment of bony resection margins (F-AICAB) in patients who had bone-infiltrating squamous cell carcinoma (SCC) of the head and neck. This was assessed to demarcate invasive carcinoma cells that stained for (cid:2)v(cid:3)6. Specimens from bony resection margins (n=362) were defined as either malignant or benign according to the results of cytological and histological examinations. Integrin (cid:2)v(cid:3)6-targeting fluorescence-labelled RGD peptides were added to the cytological samples and the accuracy of the resulting signal assessed by comparing it with the cytological findings. The value of F-AICAB was evaluated to find out if it could help to improve future diagnoses, tests, and treatments. Integrin (cid:2)v(cid:3)6 was strongly expressed in invasive SCC cells and qualified as a marker for bone-infiltrating carcinoma cells. It showed a high affinity to bind to invasive SCC cells and enabled swift and specific demarcation of (cid:2)v(cid:3)6-stained carcinoma cells. It was also diagnostic, with a sensitivity of 100% (95% CI 81.3% to 99.3%), specificity of 98.3% (95% CI 94.4% to 99.0%), positive predictive value of 92% (95% CI 70.2% to 94.3%), and negative predictive value of 100% (95% CI 96.9% to 99.9%), compared with the cytological findings. The targeting of specific integrin subtypes with selective, synthetic ligands, adapted for multimodal imaging, is a promising new approach to diagnosis. Further studies are necessary to provide more evidence for successful clinical translation and to establish the impact on clinical procedures.
关键词： Fluorescence imaging,Integrins,(cid:2)v(cid:3)6 integrin,Head and neck squamous cell carcinoma,RGD peptide,Cytology,Intraoperative margin control
更新于2025-09-04 15:30:14