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PubMed2004

Description of a flow cytometry approach based on SYBR-14 staining for the assessment of DNA content, cell cycle analysis, and sorting of living normal and neoplastic cells.

RATIONALE: This study aimed to expand the utilization of a simplified flow cytometric approach that employing SYBR-14/PI staining into broader flow cytometry applications, including (i) measurement of the DNA content; (ii) performing cell cycle analysis on mammalian cells; and (iii) sorting of live SYBR-14-stained mammalian cells based on DNA content. MATERIAL AND METHODS: Cell lines of human origin were stained with SYBR-14 and propidium iodide (PI) and assessed by a dual-color flow cytometry. Finally, sorting of living SYBR-14-stained human cell lines was performed. RESULTS: Dual staining with SYBR-14 and PI of human cells followed by flow cytometry analysis demonstrates that in addition to quality assessment, this staining could be utilized to determinate the DNA content on mammal cells. In addition, it resolves the diploid, tetraploid, and aneuploid DNA content. Furthermore, the SYBR-14-stained mammal cells were efficiently sorted based on DNA content and live cells were obtained. All these features have not been previously described with the utilization of this staining approach. CONCLUSIONS: Results of this study demonstrate that this flow cytometric approach not only allows assessment of the viability of cells, but also the DNA content of mammal cells. In addition, this approach allows one to sort viable cells stained with SYBR-14. These findings open-up unexpected and unrestricted avenues for sorting of living mammal cells and provide significant advantages over the traditionally cumbersome sorting approaches for living cells, which demand very specialized and expensive UV light sources as well as sophisticated sorting procedures.

Cell Cycle↗

PubMed1975

A comparison of subjective measurements of human sperm motility and viability with two live-dead staining techniques.

Two commonly used live-dead stains (eosin-nigrosin (EN) and eosin-opal blue (EOB)) were compared with the estimated active spermatozoa in semen samples from patients attending an infertility service. Twenty-eight semen samples were analyzed throughout the day of their collection by estimating the number of active spermatozoa and by staining a portion of the incubated sample (37 degrees C) with each stain. The samples were analyzed 30, 60, 120, 240, and 360 minutes after the initial collection. At 30 minutes there were no significant differences between the estimated values and those of either stain. The slope of the EN stain closely paralleled that of the estimated measurements throughout the remainder of the time periods, while the EOB slope was somewhat steeper. Repeatability of values for semen samples obtained on different days was generally good for each stain. A comparison of semen samples from 85 patients, 30 minutes after collection, showed no significant differences between the numbers of estimated active sperm and the percentage live using the EN stain. Studies of 132 semen samples using the EN stain showed a positive correlation with the over-all quality of sperm motility. The results indicate that there is a role for live-dead staining in assessing semen quality.

Cell Biology↗

PubMed1983

Differential staining of collagens and non-collagens with Coomassie Brilliant Blue G and R.

The following proteins were subjected to electrophoresis in SDS gels and stained with both Coomassie Brilliant Blue R and Coomassie Brilliant Blue G: the pepsin-treated collagen types I, II, III and V, and non-pepsin-treated type IV collagen, and the non-collagens, laminin, fibronectin, myosin, beta-galactosidase, fibrin, phosphorylase b and serum albumin. The Coomassie Brilliant Blue G stain was formulated as in the dye-binding protein assay reagent of Bradford (Anal. Biochem. 72: 248-254, 1976). Coomassie Brilliant Blue R prominently stained all polypeptides, but the collagen chains, including the type IV chains, stained metachromatically (red or pink) while the non-collagens stained orthochromatically (blue-violet). In the Bradford reagent, however, only the non-collagens and the intact type IV chains were prominently stained; the pepsin-treated collagen chains were virtually undetectable provided that detergent had been exhaustively removed prior to immersion in the stain. Metachromatic staining with Coomassie Brilliant Blue R is attributed to the presence of closely-spaced proline and hydroxyproline residues in sequences from triple-helical domains. The staining of type IV chains with the Bradford reagent is attributed to the presence of binding sites in the sequences from the non-triple-helical domains only, since such binding sites are absent from chains derived from the pepsin-treated collagens.

Collagen↗

PubMed1995

A picropolychrome staining technique applied to Dupuytren's tissue.

Although the histology of Duputyren's tissue is well-documented, conventional stains do not distinguish between the different types of collagen which biochemistry and immunochemistry suggest are present. Duputyren's specimens [nodules (n = 26), cords (n = 15) and dermofasciectomies (n = 6)] were stained with haematoxylin and eosin, Van Gieson's Mallory's, Masson's, and Herovici's picropolychrome stain, and examined for both cellularity and collagen staining characteristics. All stains illustrated the marked cellularity of the nodules, contrasting with a paucity of cells within the cords. The first four stains demonstrated uniformity of the collagen staining within the tissues. Herovici's picropolychrome, however, showed distinct staining patterns for the dermis, nodules and cords, with both purple/red and blue areas. Other studies suggest that those fibres stained purple/red and blue are types I and III collagens respectively. These findings may shed further light on the tissue of origin of Duputyren's disease.

Aged↗

PubMed2004

Anterior capsule staining for capsulorhexis in cases of white cataract: comparative clinical study.

PURPOSE: To compare the safety and efficacy of trypan blue 0.1%, gentian violet 0.001%, indocyanine green 0.5% (ICG), fluorescein 2%, and the patient's autologous blood for anterior capsule staining in cases of white cataract. SETTING: Rajendra Prasad Center for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India. METHODS: Fifty eyes of 50 patients with age-related white cataract had anterior capsule staining with trypan blue, ICG, or gentian violet under an air bubble or subcapsularly with fluorescein or autologous blood followed by phacoemulsification with foldable intraocular lens implantation. Each stain was used in 10 eyes. The ease of creating a continuous curvilinear capsulorhexis (CCC) and the complications during the surgery were noted. Postoperative examinations at 6 hours, 1 day, 1 week, and 1 month included slitlamp microscopy, uncorrected visual acuity, and best corrected visual acuity (BCVA). The staining patterns on the anterior capsule, side port, corneal tunnel, and anterior cortex were assessed intraoperatively and within 6 hours and at 1 day. The intraocular pressure (IOP) was assessed at 1 day; pachymetry, at 1 day and 1 month; and the endothelial cell count, at 1 month. RESULTS: The surgeon had best visualization during the anterior capsulorhexis with trypan blue, ICG, and gentian violet, and a complete CCC was achieved in all eyes in the 3 groups. Two eyes each in the fluorescein and autologous blood groups had extension of the CCC so that the capsulorhexis was complete but not curvilinear. Anterior capsule fibrosis was detected with trypan blue (1 eye) and ICG (2 eyes). The anterior vitreous was stained with fluorescein in 2 eyes. All eyes achieved a BCVA of 20/30 or better from 1 week postoperatively to the last follow-up. The side port and corneal tunnel were stained most intensely with gentian violet followed by trypan blue and ICG and less intensely with fluorescein and autologous blood. The IOP, pachymetry, and endothelial cell loss were comparable between the stains. CONCLUSION: Although trypan blue, ICG, gentian violet, fluorescein, and autologous blood were safely used to stain the anterior capsule for phacoemulsification in eyes with white cataract, trypan blue, ICG, and gentian violet were more effective in staining the capsule.

Anterior Eye Segment↗

PubMed1998

Spectrally resolved morphometry of the nucleus in hepatocytes stained by four histological methods.

A novel concept of spectrally resolved morphometry for histological specimens was developed using light microscopy combined with spectrally resolved imaging. The spectroscopic characteristics of rat hepatocytes stained by Haematoxylin and Eosin, Romanowsky-Giemsa, periodic acid-Schiff and Masson's trichrome were assessed. Light intensity in the range 450-850 nm was recorded from 10000 pixels of nuclear domains of each stained cell and represented as light transmittance spectra and optical density. In order to identify spectral shifts caused by stain-macromolecule interactions, we compared the spectra of individual stain components with those of DNA and bovine serum albumin. Chromatin and interchromatin areas were classified spectrally using a chosen spectral library followed by morphometric calculations of nuclear domains for each staining method. The spectral fingerprints of Masson's trichrome stain distinguished the nucleolus from the rest of the nuclear chromatin, enabling the demarcation and calculation of the nucleolar area. Spectrally resolved imaging of human hepatocytes stained by Masson's trichrome stain revealed marked differences between the nucleolar area in normal human hepatocytes compared with hepatocellular carcinoma. Masson's trichrome stain also distinguished the nucleolar area in human breast carcinoma cells and keratinocytes.

Animals↗

PubMed2002

Cytokeratin 7 and 20 staining for the diagnosis of lung and colorectal adenocarcinoma.

The origin of metastatic adenocarcinoma lesions can sometimes be difficult to diagnose. The objectives of our study were to establish the cytokeratin staining pattern of primary and metastatic lung and colorectal adenocarcinomas, and to determine if this helps to identify the site of origin of metastatic lesions. We reviewed a total of 102 tissue samples from patients in our tumour registry, with either primary or metastatic lung or colorectal adenocarcinoma. Tissue sections were stained for cytokeratin 7 and 20 and read as positive or negative for staining. Clinical and radiologic information was reviewed from computerised charts. The cytokeratin 7+/cytokeratin 20- pattern characterised 96% (29 out of 30) of primary and 95% (21 out of 22) of metastatic lung adenocarcinomas. All the primary (26), and 88% (21 out of 24) of metastatic colorectal adenocarcinomas stained cytokeratin 7-/cytokeratin 20+. Samples from a variety of metastatic sites were evaluated for cytokeratin 7 and 20 staining. Out of the 102 samples, in 95% (97 out of 102) of the cases, the cytokeratin 7 and cytokeratin 20 staining pattern characterised and differentiated between lung and colorectal adenocarcinoma. Primary and metastatic lung adenocarcinomas show a cytokeratin 7+/cytokeratin 20- staining pattern, while colorectal adenocarcinomas stain cytokeratin 7-/cytokeratin 20+. Cytokeratin staining is helpful in the diagnostic differentiation of metastatic lesions from these two common primaries, and assists in determining the site of origin of metastatic lesions.

Adenocarcinoma↗

PubMed2002

A rapid staining technique for the detection of the initiation of germination of bacterial spores.

AIMS: We propose to apply the Wirtz-Conklin staining technique to evaluate spore germination. METHODS AND RESULTS: Spores at different stages of germination were stained with modified spore stain (Wirtz-Conklin) and evaluated for staining properties. Bacillus spores suspended in deionized water, which does not support germination, stained greenish-blue. Spores suspended in germination enhancers that did not form bacilli stained pink, indicating the initiation of germination. Spores suspended in culture media, which promotes bacterial outgrowth, formed bacilli and were also stained pink. CONCLUSIONS: Modified spore stain (Wirtz-Conklin) was found to be useful to detect the initiation of spore germination as early as 30 min following incubation in a germination environment. SIGNIFICANCE AND IMPACT OF THE STUDY: This simple staining procedure is useful in detecting the initiation of germination of bacterial spores.

Bacillus↗

PubMed1999

Chromoendoscopy and intravital staining techniques.

Chromoendoscopy and intravital staining techniques are synonymous methods for the endoscopic early detection of malignant changes in the intestinal tract. Endoscopic intravital staining involves the use of absorptive stains (methylene blue and Lugol's solution), contrast stains (indigo carmine) and reactive stains (Congo red). Lugol's iodine solution is used to identify superficial carcinomas in the squamous epithelium of the oesophagus. Methylene blue stains the specialized intestinal epithelium in Barrett's oesophagus and, in addition to this, is helpful in the diagnosis of dysplasia. Intravital staining with indigo carmine contributes to contrasting and accentuating changed mucosal processes. Together with Cresyl violet, contrast staining is particularly important in detecting small, early malignant changes in the colon. The use of chromoendoscopy enables a biopsy diagnosis of superficial dysplastic changes and an accurate delineation of carcinomatous areas. In conjunction with the modern video-endoscopy (high-resolution endoscopy and magnification endoscopy), vital staining forms the diagnostic foundation for the detection of early malignant changes in the gastrointestinal tract. These techniques are therefore prerequisites to local endoscopic tumour therapy (mucosectomy). Despite their increasing acceptance, these methods must prove their diagnostic merit in randomized studies.

Coloring Agents↗

PubMed1991

Application of a CCD linear array camera in the quantification of tissue staining: NBT detection in ischaemic myocardium.

A method was developed to quantify the intensity of tissue staining using a CCD (charge-coupled device) camera. Reflection spectra of NBT-stained (nitro-blue tetrazolium-stained) and unstained myocardium were recorded via fibre optics coupled to a CCD camera, connected to a microcomputer. The calculation of the intensity of staining was based on evaluation of the NBT-related changes of the reflectance spectrum. In each of six anaesthetized sheep, global ischaemia was induced by cross-clamping of the aorta. The hearts were removed and incubated at 35 degrees C. At predetermined times two sections of ventricular myocardium were taken, one of which was then stained with NBT, the other being left unstained. Evaluation of the reflectance spectra from stained sections during the first phase of ischaemia showed a slight loss of NBT colour intensity followed by a more rapid loss of staining until the values of the unstained sections were reached. In contrast to the conventional visual evaluation, the method provides quantitative data on the intensity of staining, and allows use of the NBT technique for statistical evaluation of what happens over time, and the regional distribution of ischaemic injury of tissue. This method may also be applied to other staining techniques.

Animals↗

PubMed1997

Anticytokeratin antibody 34 beta E12 staining in prostate carcinoma.

Anticytokeratin antibody 34 beta E12 is advocated as an immunohistochemical stain for discriminating benign and malignant lesions of the prostate. Positive staining with 34 beta E12 is said to identify benign lesions, whereas negative staining is said to help substantiate a diagnosis of carcinoma. It is further claimed that 34 beta E12 does not stain prostate carcinoma. The studies leading to these conclusions used hematoxylin-eosin-stained sections of primary prostate lesions as controls. Although the cytokeratin content of a few cell lines of metastatic prostate carcinoma has been investigated, the 34 beta E12 immunohistochemical staining of metastatic prostate carcinoma has not been evaluated. If 34 beta E12 positivity is present only in benign prostate cells, then metastatic prostate carcinoma cells should be uniformly negative with this stain. In 14 cases of moderate and high-grade prostate cancer with metastases to lymph nodes, we found 34 beta E12 positivity in 6 (43%) of 14 metastases and in 7 (54%) of 13 primary tumors. Our findings of 34 beta E12 staining in primary and metastatic moderately and poorly differentiated prostate carcinoma differ from those reported in the literature for well-differentiated prostate carcinoma. We urge caution in the use and interpretation of 34 beta E12 staining for the diagnosis of primary and metastatic prostate carcinoma.

Adenocarcinoma↗

PubMed2000

Gram staining of protected pulmonary specimens in the early diagnosis of ventilator-associated pneumonia.

We evaluated prospectively the use of Gram staining of protected pulmonary specimens to allow the early diagnosis of ventilator-associated pneumonia (VAP), compared with the use of 60 bronchoscopic protected specimen brushes (PSB) and 126 blinded plugged telescopic catheters (PTC) obtained from 134 patients. Gram stains were from Cytospin slides; they were studied for the presence of microorganisms in 10 and 50 fields by two independent observers and classified according to their Gram stain morphology. Quantitative cultures were performed after serial dilution and plating on appropriate culture medium. A final diagnosis of VAP, based on a culture of > or = 10(3) c.f.u. ml-1, was established after 81 (44%) samplings. When 10 fields were analysed, a strong relationship was found between the presence of bacteria on Gram staining and the final diagnosis of VAP (for PSB and PTC respectively: sensitivity 74 and 81%, specificity 94 and 100%, positive predictive value 91 and 100%, negative predictive value 82 and 88%). The correlation was less when we compared the morphology of microorganisms observed on Gram staining with those of bacteria obtained from quantitative cultures (for PSB and PTC respectively: sensitivity 54 and 69%, specificity 86 and 89%, positive predictive value 72 and 78%, negative predictive value 74 and 84%). Increasing the number of fields read to 50 was associated with a slight decrease in specificity and positive predictive value of Gram staining, but with a small increase in its sensitivity and negative predictive value. The results obtained by the two observers were similar to each other for both numbers of fields analysed. Gram staining of protected pulmonary specimens performed on 10 fields predicted the presence of VAP and partially identified (using Gram stain morphology) the microorganisms growing at significant concentrations, and could help in the early choice of the treatment of VAP. Increasing the number of fields read or having the Gram stain analysed by two independent individuals did not improve the results.

Adolescent↗

PubMed1996

Staining of bone aluminium: comparison between aluminon and solochrome azurine and their correlation with bone aluminium content.

The aim of this study was to compare the sensitivity and specificity of the two histochemical stains most commonly used as indirect markers of the aluminium bone content. The clinical study was made in 28 biopsies from patients undergoing haemodialysis and the experimental study in 17 tibias from Wistar rats aluminium overloaded and with different deposits of iron. All samples were stained with aluminon, solochrome azurine and Perls and aluminium bone content was also measured. When the positive cases with Perls were excluded in the clinical study (without iron interference), the trabecular surface stained with solochrome azurine correlated with the aluminium bone content (r = 0.71, P < 0.001). With aluminon, on the other hand, no correlation was found. Solochrome azurine was always positive with aluminium contents greater than 8 micrograms/g. Aluminon was positive over 17 micrograms/g. In the experimental study, the iron concentration, in addition to other parameters, was also measured. As in the clinical study, the trabecular surface stained with solochrome azurine correlated with the aluminium content. If the positive Perls cases were excluded, the trabecular surface stained with solochrome azurine doubled the trabecular surface stained with aluminon (P < 0.001). No intratrabecular aluminon staining was observed while the intratrabecular solochrome staining correlated with the aluminium content (P < 0.001). Solochrome azurine was more sensitive than the aluminon and its lack of specificity can be easily corrected by employing Perls staining to exclude the iron interference.

Aluminum↗

PubMed2003

Grading of corneal and conjunctival staining in the context of other dry eye tests.

PURPOSE: To describe the Oxford Scheme for grading ocular surface staining in dry eye and to discuss optimization of stain detection using various dyes and filters. Also, to propose a sequence of testing for dry eye diagnosis. METHODS: The grading of corneal and conjunctival staining is described, using the Oxford Scheme, including biomicroscopy, optical filters, illumination conditions, and the characteristics of and instillation techniques used for, selected clinical dyes. RESULTS: A series of panels, labeled A-E, in order of increasing severity, reproducing the staining patterns encountered in dry eye, are used as a guide to grade the degree of staining seen in the patient. The amount of staining seen in each panel, represented by punctate dots, increases by 0.5 of the log of the number of dots between panels B to E. The use of the vital dyes fluorescein, lissamine green, and rose Bengal is described; fluorescein and lissamine green, used in conjunction with appropriate absorption filters, are recommended for use in clinical trials. The placement of staining in relation to the sequence of other diagnostic tests is discussed. CONCLUSIONS: The monitoring and assessment of corneal and conjunctival staining can be greatly enhanced by the use of a grading scale, controlled instillation of dyes, and standard evaluation techniques. This is of particular benefit in clinical trials, where ocular surface staining is commonly employed as an outcome measure

Coloring Agents↗

PubMed1996

The response of selected members of the archaea to the gram stain.

Archaea possess a broader range of cell envelope structural formats than eubacteria and their cell walls do not contain peptidoglycan. Some archaea have only a single S-layer as their cell wall (e.g. Methanococcus jannaschii and Sulfolobus acidocaldarius), whereas others have multiple layers (e.g. Methanospirillum hungatei). Sometimes there can also be a high proportion of tetraether lipids in membranes to make the envelope more resilient to environmental stress (e.g. Methanococcus jannaschii and Sulfolobus acidocaldarius grown at 70 degrees C). Since the Gram reaction depends on both the structural format and the chemical composition of the cell envelope of eubacteria, it was important to determine if the same is true for archaea. Methanospirillum hungatei, Methanosarcina mazei, Methanobacterium formicicum, Methanococcus jannaschii and Sulfolobus acidocaldarius, chosen because of their different envelope formats and chemistries, were subjected to a Gram stain that can be used for transmission electron microscopy. In this staining regimen, the iodine is replaced by potassium trichloro(eta 2-ethylene)platinate(II) as the mordant, and the platinum of the new compound is the electron-scattering agent for electron microscopy. Of all these archaea, only Methanobacterium formicicum stained Gram-positive since its pseudomurein wall remained intact; the platinum compound formed large electron-dense aggregates with the crystal violet that were located in the vicinity of the cell wall and the plasma membrane. All but the terminal filament cells of Methanospirillum hungatei stained Gram-negative because the limiting porosity of its external sheath was so small that the Gram reagents could not enter the cells. The terminal cells of filaments stained Gram-positive because the staining reagents gained entry through the terminal plugs. All other archaea stained Gram-negative because their cell walls were so disrupted during staining that the crystal violet-platinum complex could not be retained by the cells. Methanococcus jannaschii was grown at both 50 degrees C and 70 degrees C so that the tetraether lipids in its plasma membrane could be increased from 20% (50 degrees C) to 45% (70 degrees C) of the total lipids; in both cases the cells stained Gram-negative.

Archaea↗

PubMed2005

Cluster designation 5 staining of normal and non-lymphoid neoplastic skin.

BACKGROUND: Immunohistochemical staining for cluster designation 5 (CD5) has been found to label a variety of non-lymphoid tumors. METHODS: A variety of eccrine, apocrine, follicular, epithelial, and pagetoid lesions were selected and stained with an anti-CD5 monoclonal antibody (Novocastra Labs, Newcastle upon Tyne, UK, clone 4C7) by immunohistochemistry. The intensity of positive cytoplasmic staining was graded semiquantitatively (1+ weak staining, 2+ strong staining). Additionally, the percentage of positive lesional cells was placed in one of four categories: >75%, 25-75%, 1-25%, and <1%. RESULTS: Within normal skin, CD5 labeled lymphocytes, apocrine glands, deep dermal eccrine glands, and smooth muscle (weak). The majority of benign and malignant apocrine lesions demonstrated strong focal (36%, n=11)-to-diffuse (64%, n=16) staining. In contrast, labeling of benign eccrine tumors was more focal, tending to localize around ducts (79%, n=19). Microcystic adnexal carcinoma demonstrated focal staining of deeper ductal structures (71%, n=7), whereas desmoplastic trichoepithelioma and basal cell carcinoma showed only rare positive cells. All cases of mammary (n=7) and extramammary (n=8) Paget's disease labeled diffusely for CD5. Pagetoid Bowen's disease (n=6), intraepidermal sebaceous carcinoma (n=3), nor melanoma in situ (n=6) showed any CD5 staining. CONCLUSIONS: Immunohistochemical staining for CD5 is extremely useful in the differential diagnosis of pagetoid epidermal lesions and will mark mammary and extramammary Paget's disease, but not pagetoid Bowen's disease, melanoma in situ, or sebaceous carcinoma.

CD5 Antigens↗

PubMed2003

Effect of insulin on rat ovarian leptin expression by immunohistochemical staining.

AIM: Leptin and insulin may interact in regulating ovarian steroid synthesis. The objective of this study was to investigate immunohistochemical staining of leptin in normal rat ovarian tissues and in rats treated with insulin and insulin plus human chorinoic gonadotropin (hCG). METHODS: Paraffin blocks of rat ovarian tissues from a previous study, in which 18 adult, female Wistar rats with an average weight of 250 g were divided into three groups to receive either saline solution, human insulin (2 U/day) or human insulin (2 U/day) plus hCG (4 U/day) for 4 weeks, were used in this study to compare the effects on leptin staining. The results were analysed using a semiquantitative scoring system, such as mild, moderate and strong. RESULTS: No staining was observed in granulosa cells and theca interna cells of normal ovarian tissues. Theca externa cells had mild staining intensity (+), corpus luteum had moderate (+ +) and stroma had mild (+) staining intensity. Histological structure was impaired in the insulin group, luteinized cells had mild staining, there was no difference in other cell groups. Only theca externa cells of the developing follicles were stained in insulin plus hCG group, luteinized cells again had mild staining. CONCLUSIONS: Besides damaging the rat ovarian structure, insulin reduced staining intensity of leptin in luteinized cells. Insulin may stimulate ovarian steroid synthesis not only through its own receptors, but also by acting on the leptin expression of these cells.

Animals↗

PubMed2005

Rapid toluidine blue stain for Mohs' micrographic surgery.

BACKGROUND: Toluidine blue is a useful stain for detecting basal cell carcinoma during Mohs' micrographic surgery. OBJECTIVE: To demonstrate the efficacy of alkalinization on the toluidine blue stain. METHODS: A 1% aqueous toluidine blue-1% aqueous sodium borate solution was used to stain microscope slides for basal cell carcinoma during Mohs' micrographic surgery. RESULTS: Total toluidine blue staining time was reduced to less than 2.5 minutes, without compromising the quality of the stain. CONCLUSIONS: The rapid toluidine blue stain reduces staining time while maintaining staining quality, including the advantages specific to the toluidine blue stain.

Basal Cell Carcinoma↗