Ultrafast Toluidine Blue Staining for Rapid On-Site Evaluation of Cytological Smears

Hewer, Ekkehard; Schmitt, Anja

doi:10.1159/000505254

Cited by 7 publications

(10 citation statements)

References 6 publications

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“…Traditional methods like Papanicolaou and Giemsa stains, despite their diagnostic clarity and established utility, pose challenges in terms of time and resource requirements [ 9 , 12 , 13 , 15 ]. This has catalyzed the exploration of alternative staining methods, as evidenced by the research conducted by Amannagi et al [ 16 ] and Hewer et al [ 17 ] and our own previous investigation [ 12 ].…”

Section: Discussionmentioning

confidence: 99%

“…Their findings underscore the importance of rapid staining techniques, highlighting how such methods can facilitate prompt preliminary assessments without compromising diagnostic accuracy. Similarly, Hewer et al [ 17 ] also investigated toluidine blue in thyroid FNAB, emphasizing its efficiency in cytology reporting. However, similar to Amannagi’s study [ 16 ], Hewer et al [ 17 ] concentrated primarily on sample adequacy rather than a comprehensive cytological evaluation.…”

Section: Discussionmentioning

confidence: 99%

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Evaluating Diagnostic Clarity: The Comparative Efficacy of BlueStain in Serous Effusion Cytology under the International System for Reporting Serous Fluid Cytopathology Reporting Framework

Alves,

Azevedo,

Ferreira

et al. 2024

Diagnostics

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Serous effusion cytology is a pivotal diagnostic and staging tool in clinical pathology, valued for its simplicity and cost-effectiveness. Staining techniques such as Giemsa and Papanicolaou are foundational, yet the search for rapid and efficient alternatives continues. Our study assesses the efficacy of an in-house-developed BlueStain, a toluidine blue variant, within the International System for Reporting Serous Fluid Cytopathology (TIS), aiming to optimize diagnostic clarity and resource use. Materials and Methods: This section provides details on the cohort of 237 patients with serous effusions, the ethical approval process, sample collection, and staining procedures with BlueStain, Papanicolaou, and Giemsa. It also describes the microscopic evaluation criteria, scoring system, and statistical methods used to compare the stains. Results: BlueStain demonstrated notable performance, particularly in identifying malignant cells, presenting a competitive alternative to the Papanicolaou stain, which, despite higher quality indices in other categories, requires more resources and time. The study revealed that BlueStain might offer a valuable balance between quality and efficiency, especially in cases where rapid diagnostic turnaround is essential. Conclusions: Our findings suggest that BlueStain is a viable staining method in the context of serous effusions, capable of providing detailed cytomorphological analysis. While traditional stains hold their place for their established diagnostic clarity, BlueStain offers a rapid and resource-optimized alternative. The absence of definitive diagnostic criteria in the atypical category and the inherent sample heterogeneity underscores the necessity for adaptable staining methods like BlueStain. The study highlights the potential trade-offs between detail and practicality in staining techniques, advocating for further research into innovative methods that do not compromise diagnostic precision for cost and time efficiency.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Evaluating Diagnostic Clarity: The Comparative Efficacy of BlueStain in Serous Effusion Cytology under the International System for Reporting Serous Fluid Cytopathology Reporting Framework

Alves,

Azevedo,

Ferreira

et al. 2024

Diagnostics

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“…There are some factors that make a rapid stain widely accepted for use. A rapid stain should provide an easy and successful fixation, or preferably, no requirement of fixation at all, cellular transparency, an easy and user-friendly staining process, and low cost [4,5]. For the most common rapid staining method, DQ, the entirely air-dried smears are "must-have" to provide a successful staining.…”

Section: Discussionmentioning

confidence: 99%

“…A good and correct adequacy assessment affects the success of a final report in cytopathology. There are a few stains available in the market for ROSE, namely, Diff-Quik (DQ), Toluidine blue, and ultrafast Papanicolaou stains [4,5]. Of those, the most common in use is DQ [6].…”

Section: Introductionmentioning

confidence: 99%

A New Cytology Staining Method: A Fast Approach for Rapid On-Site Evaluation on Thyroid Fine-Needle Aspiration Cytology

Alves¹,

Ferreira²,

Oliveira³

et al. 2022

Acta Cytologica

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Introduction: Thyroid fine-needle aspiration (FNA) is a well-established technique in the cytology literature. Through the introduction of rapid stains in cytology practice, the ever-increasing utility of rapid on-site evaluation (ROSE) has strengthened the place of FNA as a primary diagnostic method in patient management. There are few stain variants available in the market for ROSE, namely Diff-Quik (DQ), Toluidine blue, and ultrafast Papanicolaou stains. Recently, our group developed a new staining variant labeled as original “BlueStain®” technique that was not previously tested in this context. Methods: 40 FNA thyroid cases were studied. At least two slides were prepared from each patient: one stained by DQ and the other by BlueStain®. Simultaneously, a ROSE diagnosis was performed as the two staining methods were compared, evaluating the parameters of background, cellularity, details of colloid presence, cell morphology, nuclear details, cytoplasmic details, and overall staining, scored on a scale from 1 to 3, representing poor, average, and good, respectively. Results: The quality index was slightly better for BlueStain® (53% vs. 47%) but not significantly different between the two stains. BlueStain® provides better details in both the presence and type of colloid as well as nuclear details, which are regarded as very important for diagnosis in thyroid cytology. There were eight cases with discordant diagnosis when compared between two stains from the same patient. In five cases of indeterminate cases, BlueStain® allows to bring them to the benign category, probably because this staining method allows a clear observation of the colloid in the background of the smears. However, since we are observing two different slides, we cannot rule out that these differences are a question of sample collecting and/or smearing. Conclusions: Our data demonstrates that BlueStain® is suitable to provide good-quality slides for primary assessments of thyroid aspirates studied by ROSE. In fact, in some aspects, this new staining method shows better preservation of colloid and cell details, revealing itself as an alternative to the DQ stain variant, upholding performance level while being 10 times cheaper and simpler because it requires just one step of staining.

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“…The excess dye was removed and the cells were washed thrice with PBS (2 min/step). The cells were imaged using an optical microscope (XDS-1A) and an inverted microscope (Olympus, IX71) ( Hewer and Schmitt, 2020 ).…”

Section: Methodsmentioning

confidence: 99%

Effect of Amniotic Membrane/Collagen-Based Scaffolds on the Chondrogenic Differentiation of Adipose-Derived Stem Cells and Cartilage Repair

Cao

Tong

Wang³

et al. 2021

Front. Cell Dev. Biol.

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Objectives: Repairing articular cartilage damage is challenging. Clinically, tissue engineering technology is used to induce stem cell differentiation and proliferation on biological scaffolds to repair defective joints. However, no ideal biological scaffolds have been identified. This study investigated the effects of amniotic membrane/collagen scaffolds on the differentiation of adipose-derived stem cells (ADSCs) and articular cartilage repair.Methods: Adipose tissue of New Zealand rabbits was excised, and ADSCs were isolated and induced for differentiation. An articular cartilage defect model was constructed to identify the effect of amniotic membrane/collagen scaffolds on cartilage repair. Cartilage formation was analyzed by imaging and toluene blue staining. Knee joint recovery in rabbits was examined using hematoxylin and eosin, toluidine, safranine, and immunohistochemistry at 12 weeks post-operation. Gene expression was examined using ELISA, RT-PCR, Western blotting, and immunofluorescence.Results: The adipose tissue was effectively differentiated into ADSCs, which further differentiated into chondrogenic, osteogenic, and lipogenic lineages after 3 weeks’ culture in vitro. Compared with platelet-rich plasmon (PRP) scaffolds, the amniotic membrane scaffolds better promoted the growth and differentiation of ADSCs. Additionally, scaffolds containing the PRP and amniotic membrane efficiently enhanced the osteogenic differentiation of ADSCs. The levels of COL1A1, COL2A1, COL10A1, SOX9, and ACAN in ADSCs + amniotic membrane + PRP group were significantly higher than the other groups both in vitro and in vivo. The Wakitani scores of the ADSC + amniotic membrane + PRP group were lower than that in ADSC + PRP (4.4 ± 0.44**), ADSC + amniotic membrane (2.63 ± 0.38**), and control groups (6.733 ± 0.21) at week 12 post-operation. Osteogenesis in rabbits of the ADSC + amniotic membrane + PRP group was significantly upregulated when compared with other groups. Amniotic membranes significantly promoted the expression of cartilage regeneration-related factors (SOX6, SOX9, RUNX2, NKX3-2, MEF2C, and GATA4). The ADSC + PRP + amniotic membrane group exhibited the highest levels of TGF-β, PDGF, and FGF while exhibiting the lowest level of IL-1β, IL6, and TNF-α in articular cavity.Conclusion: Amniotic membrane/collagen combination-based scaffolds promoted the proliferation and cartilage differentiation of ADSCs, and may provide a new treatment paradigm for patients with cartilage injury.

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Ultrafast Toluidine Blue Staining for Rapid On-Site Evaluation of Cytological Smears

Cited by 7 publications

References 6 publications

Evaluating Diagnostic Clarity: The Comparative Efficacy of BlueStain in Serous Effusion Cytology under the International System for Reporting Serous Fluid Cytopathology Reporting Framework

Evaluating Diagnostic Clarity: The Comparative Efficacy of BlueStain in Serous Effusion Cytology under the International System for Reporting Serous Fluid Cytopathology Reporting Framework

A New Cytology Staining Method: A Fast Approach for Rapid On-Site Evaluation on Thyroid Fine-Needle Aspiration Cytology

Effect of Amniotic Membrane/Collagen-Based Scaffolds on the Chondrogenic Differentiation of Adipose-Derived Stem Cells and Cartilage Repair

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