Staining Technology Research Articles

Genetic diversity of DNA microsatellite for Tibetan Yak

To assess the genetic diversity and relationship of the Tibetan yak breeds. The genetic diversity and phylogenies of a total of 480 individual from 11 Tibetan yak groups were analyzed using PCR and multiplex gel electrophoresis of silver staining technology with eight pairs of microsatellite markers.The result showed that these markers were highly polymorphic loci with rich genetic diversity in the Tibetan yak populations.The average polymorphic information content (PIC) in 11 groups of yak were higher than 0.5. The highest HEL13 was 0.8496, and the lowest TGLA57 was 0.7349. Among them, the PICof Dingqing yak was minimum (0.7505), indicating that the group is relatively pure.Sangri Yak had the highest PIC value (0.7949) indicating greater genetic variationwithinthe groups. Among the 11 groups examined, the order of heterozygosity size wasSangri(0.8193)>Jiangda(0.8190)>Sangsang(0.8157)>Baqing(0.8150)>Kangbu(0.8123)> Jiali(0.8087)>Gongbujiangda(0.8054)>Sibu(0.8041)>Leiwuqi(0.8033)>Pali(0.8031)>Dingqing(0.7831). The groups from eastern Tibet had grater genetic diversity than those from Western Tibet, which indicate that Tibet may be one of the cradles of the yak.According to the genetic distance, the cluster relationship constructed with UPGMA and NJ methods showed that 11 yak groups in Tibet could be divided into three forms. In summary,Tibet yak has abundant genetic diversity and the selected microsatellite markers can be used to evaluategenetic diversity of Tibetan yak.

Characterization of glioblastoma (GBM) vasculature and protein expression of surrounding tumor cells on single FFPE sections with a multicycle multiplexed in situ immunofluorescent staining technology.

2097 Background: GBM is the most common brain tumor in humans and has a dismal prognosis. Although antiangiogenic therapy (bevacizumab) is an option for GBM, there is still unmet need to understand tumor pathophysiology and predictive biomarkers. We built a tissue based multiplexed immunofluorescent assays and developed algorithms to identify and quantify tumor vasculature, that enabled quantification, visualization, and colocalization of multiple protein in surrounding tumor cells at single cell and subcellular levels. This assay provides unique opportunity to explore tumor heterogeneity of tissue morphology and their relationships to vasculature, and is a novel tool for biomarker and treatment discovery. Methods: Tissue micro arrays (TMAs) were constructed from 141 GBM patients. Fluorescent dye labeled antibodies against 18 biomarkers were sequentially applied on single sections of these TMAs. Metrics were built to identify vessels, quantify distance of tumor cells to vessels, and analyze expression profiles of biomarkers, including signaling molecules in EGFR, PI3K/AKT, TGF-beta pathways, and hypoxia marker Glut1, in proximity to blood vessels. Results: CD31 was successfully used to identify blood vessels in GMB. Vessel segmentation and quantification were performed on all of the images. Biomarker profiling in the context of blood vessels demonstrated different patterns in close proximity to vessels, with some biomarkers showing increased levels (e.g. SMA, EGFR, pS6), some showing decreased levels (e.g. p4EBP), and others remain the same (FOXO3a, S6). Quantification of biomarkers showed heterogeneous expression within the same sample and across the cohort. In addition, co-localization of the above markers was visualized and demonstrated on single cell and subcellular levels. Conclusions: We were able to use a novel fluorescent multiplexing technology (MultiOmyx) to study GBM biology. This technology allowed the simultaneous analyses of multiple biomarkers of GBM, and provides new insights on the relationship of markers to each other, tumor heterogeneity and angiogenesis.

Nucleolar activity of haemocytes in the adult firebug as an alternative animal model

Firebug haemocytes are a newly developed alternative animal model, but data on their physiology are deficient. We examined the nucleolar activity, which reflects the intensity of cellular proteosynthesis, using toluidine blue and AgNOR stain technology. Prohaemocytes, granulocytes and plasmatocytes contain on an average two nucleoli, whereas spherulocytes have on average six. The number of compact nucleoli, which are the attribute of intensive proteosynthetic activity, was significantly higher in prohaemocytes. Our data support the hypothesis that prohaemocytes have a physiology similar to mammalian haematopoietic stem cells.

Abstract 1857: Intracellular protein GP128 is a novel tumor angiogenesis marker on gastric cancer cell lines and GI tissues: Ectopic expression on the cell surface defined by MS38-2.1 mAb

Abstract Our aim is to generate monoclonal antibodies (mAbs) against novel markers on the cancer cell surface using an innovated high throughput screening (HTS) strategy. Mixed live cells from four gastric cancer cell lines (MKN45, SGC7901, BGC823 and MKN28) were used as the immunogen for A/J mice: spleen cells were then fused with mouse SP2/0 myeloma cells to generate monoclonal antibody hybridomas. The 16 highly positive hybridoma colonies were selected from 20,000 colonies in sixty-seven 96-well plates through screening using live cell (gastric cancer cells versus normal PBMC) staining and FACS-HTS technology. Protein-A affinity purified mAbs were used to define the antigens, which were confirmed as novel biomarkers on the cancer cell surface using microarray, mass and proteomic analyses. Protein microarray analysis with the MS38-2.1 mAb identified an intracellular protein GP128 (e.g., SPRR3) as the most likely a novel cancer biomarker. The immunohistochemical profiles of MS38-2.1 mAb in GI (gastric and colorectal) cancer tissues were found to be significantly higher (P<0.01) than those in normal GI tissues, and confirmed to be restricted to the small- and medium-sized blood vessels and the cancer cells. Human SPRR3 belongs to a family of small proline-rich repeat proteins that is normally expressed intracellularly. Our findings demonstrate that GP128, like SPRR3, could be ectopically expressed in the extracellular matrix in GI cancers. Conclusion: Our results demonstrate that the intracellular protein GP128 might play a role in gastrointestinal cancer development. This investigation also provides an example of identification of potential cancer biomarkers through a novel screening technique. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1857. doi:1538-7445.AM2012-1857

A new DNA band display technology of microsatellite DNA

This study proposes a new DNA band display technology of microsatellite DNA called Fluorescent Imaging Technology. In comparison with Silver Stain Technology, this technology is worth popularizing in the laboratory because of its high resolution, efficiency, simplicity and clear background. Key words : Microsatellite DNA, polyacrylamide gel electrophoresis, fluorescent imaging technology, silver staining technology.

The detection of a transgenic soybean biochip using gold label silver stain technology

A method for the rapid detection of transgenic soybean crops based on a combination of gene chip and "gold label silver stain" (GLSS) technologies has been established. To ensure the specificity of this method, the CaMV35S promoter and Nos terminator were selected as probes because they are both exogenous genes that are specific to transgenic soybean plants. The addition of biotin-modified dUTPs to a polymerase chain reaction (PCR) system can produce amplified nucleic acid segments containing biotin. These labeled PCR products then hybridize with specific probes on the chip and are subsequently bound by streptavidin-modified gold nanoparticles (GNPs). Due to the catalytic nature of the GNPs, silver staining can be used to visualize the hybridized probes, which appear as signals in varying shades of gray. The intensity value of the gray signals can be obtained using a general scanner. Silver staining for 10 min was determined to produce the optimal signal-to-noise ratio. In addition, this method was shown to be highly specific and had a detection sensitivity of 288.57 pg/μL.

Screening for cervical neoplasia: A community‐based trial comparing Pap staining, human papilloma virus testing, and the new bi‐functional celldetect® stain

Although cytological screening for cervical neoplasia has lowered mortality rates, current screening methods are plagued by sub-optimal sensitivity and/or specificity. The purpose of this study was to compare the performance of the new CellDetect® staining technology as a potential screening tool. This initial, non-blinded study, utilized samples are taken at a community-based clinic. The diagnostic results using CellDetect® were compared with the performance of Pap staining and human papilloma virus (HPV) testing on the same material, as well as the follow-up biopsies. These data were statistically analyzed in terms of sensitivity, specificity, predictive value (N.P.V and P.P.V), and inter-observer agreement. Bi-functional CellDetect® staining revealed morphological details and tinctorial properties that permitted recognition of neoplasia even at low magnification. Performance-wise, CellDetect® demonstrated non-inferiority for all statistical parameters to both Pap and HPV tests. Importantly, superior sensitivity compared with Pap staining was observed, as well as higher specificity than HPV testing with near equivalent sensitivity. We conclude that CellDetect® is a promising approach to early detection of cervical cancer because of its bi-functional capabilities that afford high sensitivity and specificity. The data suggest that this new methodology warrants further and more extensive clinical evaluation.

Abstract 4155: Simultaneous quantitation of multiple signaling molecules in individual circulating tumor cells (CTCs) by multi-color laser scanning cytometry

Abstract Introduction: Recent advances in circulating tumor cell (CTC) technology provide a new alternative approach to examine proteomic and molecular characteristic of tumor cells from cancer patients. The current trend in anti-cancer drug development focuses on new compounds that modulate multiple signaling pathways. With CTC detection used as a potential liquid biopsy, the simultaneous examination of signaling molecules for multiple pathways on the same CTC could provide insight into the drug activity in tumor cells. We used proprietary laser scanner cytometry (LSC) technology and developed a multi-color immunofluorescence (IF) procedure to detect 7 biomarkers within the same CTC. We validated this new staining technology using the (1) receptor tyrosine kinase pathways based molecules included total and phospho-IGF-1R, in combination with total and phospho-EGFR, or with total and phospho-ERK1/2, or with total and phospho-AKT (S473) in HT-29 colorectal tumor cells, and (2) same marker expression in CTCs enriched from cancer patients. Purpose: To validate the new multiple marker IF staining procedure and laser scanning cytometry (LSC) platform using ligand treated or untreated tumor cells, and in CTCs enriched from cancer patients. Method: A target biomarker staining panel was combined with standard CTC staining (DAPI, CK+ and CD45-) to create a 7-color IF staining panel to examine marker expression in tumor cell lines and in CTCs from cancer patients. Cultured tumor cells and CTCs enriched from Cellsearch Profile were cytospun onto slides, stained, scanned (by LSC) and analyzed. Results: Using a simultaneous co-staining procedure and LSC-based analysis, we demonstrated that treatment of HT29 cells with IGF1 for 15-30 minutes induced significant increases in both p-IGF1R expression and p-ERK1/2 and p-AKT expression on HT29 cells. In addition, anti-IGF1R and/or anti-EGFR, or with multi-kinase inhibitor treatment leads to differential reduction of p-ERK1/2 and p-AKT. Using CTCs enriched from cancer patient, simultaneous detection of multiple signaling molecules was also observed. Summary A multi-color, LSC-based methodology was developed to quantify expression of up to 4 biomarkers in CTCs. Application of this technology in clinical studies to examine the activation status of CTCs from patients could provide important information on pathway-specific drug activity and guide dosing selection. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4155. doi:10.1158/1538-7445.AM2011-4155

Abstract 5172: Fluorescence labeling of cytokeratin-negative circulating tumor cells in peripheral blood: A new paradigm for the study of cancer progression

Abstract Introduction: The measurement of circulating tumor cells (CTCs) in blood generally requires either anti-EpCAM for capture, anti-cytokeratin (CK) for detection, or both. However, EpCAM and CK are absent in some tumor cells, and both may be down-regulated during neoplastic progression or during epithelial-to-mesenchymal transition (EMT). Here we show capture on a micro-fluidic system using single or multiple antibodies, and CTC detection with CEE-Enhanced (CE), a novel in situ staining method that fluorescently labels the capture antibodies bound to the CTCs. Methods: Buffy coat cells isolated from 8 mL of blood were pre-incubated with either anti-EpCAM alone or with a mixture of antibodies to epithelial and mesenchymal cell surface antigens. CTCs were captured on a micro-fluidic channel. Identification of CTCs was determined with anti-CK and with CE to detect cells with the capture antibodies bound to the cell surface antigens. All cells scored as positive for CK or CE were, by definition, CD45-negative and DAPI-positive. Results: Using anti-EpCAM alone for capture, significantly more CTCs were detected by CE staining than with anti-CK in breast and prostate cancers. This indicated that CK-negative CTCs were captured but not detected and that some EpCAM-positive CTCs were CK-negative. Results using capture antibody mixtures varied from sample to sample but gave up to 2-fold higher CK-positive cells than anti-EpCAM only, and as much as 4-fold higher CE-positive cells. Control blood from healthy donors was CK and CE-negative. All CK-positive cells co-stained with CE, as determined with different fluorescent labels. In a clinical study of stage IV breast cancer, CTCs were isolated with an antibody mixture and sequentially stained with CK and CE. Fifteen of 24 samples (63%) contained CK-positive cells (range 1-60 CTCs) while 24 of 24 samples (100%) contained additional CE-positive cells (range 1-41; median=11; Wilcoxon test, p=0.02). The modest correlation coefficient (r = 0.57, p=0.004) for the number of CE-positive and CK-positive cells in each sample suggests that one or more different phenotypes of CTCs were being detected. Amplified Her2 was detected by FISH in isolated CK-positive CTCs, and also in CK-negative, CE-positive CTCs from Her2-positive patients, indicating these were tumor cells. Conclusions: The CEE-Enhanced staining technology enables the detection of CK-negative CTCs. This novel detection method, based on the in situ labeling of antibodies used for capture, greatly expands single and multi-antibody approaches to the study of rare circulating cells. Specifically, this allows the exploration and study of circulating cells not expressing CK such as highly de-differentiated tumor cells, stem cells or those tumor cells undergoing EMT. The clinical significance of these CK-negative CTCs is under investigation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5172. doi:10.1158/1538-7445.AM2011-5172

Advances in the Analysis of Hepatitis C Virus Specific T Cell Responses

There is a growing consensus that cellular immune responses are associated with the clinical outcome of hepatitis C virus infection (HCV). The development of Tetramer staining, ELISpot, flow cytometry and epitope mapping technologies make it possible to enumerate, phenotype and assess the proliferation and function of HCV specific T cells; as well as map and predict T cell epitopes and track the evolution of T cell epitopes. Such information is essential for the development of effective therapeutic and prophylactic HCV vaccines. This article summarizes the technical advances relevant to HCV specific T cell responses.

COMPARATIVE ANALYSIS OF DIRECT FLUORESCENCE, ZENON LABELING, AND QUANTUM DOT NANOCRYSTAL TECHNOLOGY IN IMMUNOFLUORESCENCE STAINING

A comparative analysis was performed to determine the sensitivity and efficiency of three fluorescent labeling techniques, including direct fluorescent-antibody staining (FA), Zenon labeling, and quantum dot (QD) nanocrystal technology. Two varicella-zoster virus immunoglobin (Ig) G forms, mAb 4F9 and mAb g62, were selected for these studies. The results indicated that: (1) All three methods demonstrated similar brightness and photostability; (2) the time required to conjugate the antibody varied, with Zenon labeling being the quickest; and (3) the stability of each conjugated complex was different, with FITC/rhodamine-conjugated antibody being the most stable.

“Microenvironmental contaminations” induced by fluorescent lipophilic dyes used for noninvasive in vitro and in vivo cell tracking

AbstractDetermining how normal and leukemic stem cells behave in vivo, in a dynamic and noninvasive way, remains a major challenge. Most optical tracking technologies rely on the use of fluorescent or bioluminescent reporter genes, which need to be stably expressed in the cells of interest. Because gene transfer in primary leukemia samples represents a major risk to impair their capability to engraft in a xenogenic context, we evaluated the possibility to use gene transfer–free labeling technologies. The lipophilic dye 3,3,3′,3′ tetramethylindotricarbocyanine iodide (DiR) was selected among 4 near-infrared (NIR) staining technologies. Unfortunately we report here a massive transfer of the dye occurring toward the neighbor cells both in vivo and in vitro. We further demonstrate that all lipophilic dyes tested in this study (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindotricarbocyanine perchlorate [DiI], DiD, DiR, and PKH26) can give rise to microenvironmental contamination, including when used in suboptimal concentration, after extensive washing procedures and in the absence of phagocytosis or marked cell death. This was observed from all cell types tested. Eventually, we show that this microenvironmental contamination is mediated by both direct cell-cell contacts and diffusible microparticles. We conclude that tracking of labeled cells using non–genetically encoded markers should always be accompanied by drastic cross validation using multimodality approaches.

Histological Analysis of Single Peripheral Nerve Fiber in Acute Nerve Elongation Process

To observe the histological alterations of single nerve fiber structures after nerve elongation by employing a rabbit peroneal nerve stretching model. 14 rabbits weighing mean 3. 0 kg (2.02-3.31 kg.) were used in the experiment. Two rabbits were used as control when only a sham operation was done (group one, 0% stretch). Acute stretching of the peroneal nerves to elongate them by 10% was done in 6 rabbits (group two, 10% elongation) and by 20% (group three, 20% elongation) in another 6 rabbits. All animals were evaluated by tissue staining technology in a teased-fiber study. The internodal lengths were measured, and nodes of Ranvier and Schmidt-Lanterman notch were observed. The nerve fiber length was increased after stretching. The mean internodal length was 1208.31 microm in group one, 1347.26 microm in group two, and 1411.35 microm in group three. Compared with the control group, mean internodal length was elongated by 11.50% in group two and 16.80% in group three. The difference was statistically significant. The node of Ranvier and Schmidt-Lanterman notch was wider in both group two and group three. Rupture of nerve fiber at the node of Ranvier was observed in group three. The peroneal nerve in rabbits can adapt to mild stretching by internodal length elongation. Elongation by 20% will cause structural rupture and therefore is the limit for nerve elongation.

Research on a Novel Rigid Internal Fixation System

The purpose of the present study was to evaluate the efficiency of fixation with a novel (bulge core type system) rigid internal fixation system. Mandibular linear fractures were created artificially in dogs and fixed using a bulge core type system or a miniplate system in vitro. A 3-point load cell was used to contrast the fixation strength of the 2 systems in vitro. Histologic sections were used to analyze the healing of mandibular linear fractures and to contrast the efficiency of fixation of the 2 systems in vivo. Tetracycline-fluorescein staining technology was used to evaluate the bone healing speed in the mandibular fracture lines during bone healing. In vitro, the 3-point load cell showed that the fixation strength of the bulge core type system was greater than that of the miniplate system (P < .05). In vivo, the histologic sections and tetracycline-fluorescein staining showed that greater and quicker bony healing occurred using the bulge core type system (P < .05) and that the efficiency of fixation of the 2 systems was similar. Our results have shown that the bulge core type system can provide sufficient strength for mandibular linear fracture healing.

Protein subcellular localization profiling of breast cancer cells by dissociable antibody microarray staining

We have developed dissociable antibody microarray (DAMA) staining technology that provides a new approach to the global analysis of protein subcellular localization (SCL) in fixed cells. We have developed and optimized this technology for protein SCL profiling, generated ChipView, a program for management and analysis of molecular image database, and utilized the technique to identify proteins with unique SCL in breast cancer cell lines. We compared the SCL profiles of 325 proteins among nine different breast cell lines, and have identified one protein, Cyclin B1, with distinctively different SCLs between normal and cancer cell lines. With classic individual immunostaining, Cyclin B1 was confirmed to localize to the cytoplasm of seven breast cancer cell lines and in both cytoplasm and nuclei of two normal breast cell lines, and to have higher expression levels in the cancer cell lines tested.

High frequency of spontaneous triploids in barley

HereditasVolume 64, Issue 1 p. 131-134 Open Access High frequency of spontaneous triploids in barley Jens Sandfaer, Jens SandfaerSearch for more papers by this author Jens Sandfaer, Jens SandfaerSearch for more papers by this author First published: February 1970 https://doi.org/10.1111/j.1601-5223.1970.tb02280.xCitations: 2AboutPDF ToolsExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Literature cited Derenne, P. 1967. Triploïdie et trisomie chez l'orge cultivée, I. L'état triploïde. Bull. Rech. Agron. Gembloux N. S. 2: 242– 259. Jacobsen, P. 1954. Chromosome numbers in the genus Hedera L. Hereditas 40: 252– 254. Jacobsen, P. 1965. An improved squash-slide freezer. Stain Technol. 40: 63– 65. Kerber, E. R. 1954. Trisomics in barley. Science 120: 808– 809. Nilan, R. A. 1964. The cytology and genetics of barley 1951–1962.—Monogr. Suppl. No. 3, Res. Studies 32, No. 1. Wash. State Univ.. Sandfaer, J. 1968. Induced sterility as a factor in the competition between barley varieties. Nature 218: 241– 243. Sandfaer, J. 1970. Barley stripe mosaic virus as the cause of sterility interaction between barley varieties. Hereditas 64: 150– 152. Smith, L. 1951. Cytology and genetics of barley. Bot. Rev. 17: 1– 355. Tsuchiya, T. 1958. Studies on cross compatibility of diploid, triploid and tetraploid barleys I. Seed development in reciprocal crosses between diploids and induced autotetraploids. Bot. Mag. Tokyo 71: 48– 52. Tsuchiya, T. 1960. Studies on cross compatibility of diploid, triploid and tetraploid barley, III. Results of 3x × 3x crosses. Seiken Zihô 11: 29– 37. Citing Literature Volume64, Issue1February 1970Pages 131-134 ReferencesRelatedInformation

Repetitious DNA in mammalian chromosomes.

HereditasVolume 69, Issue 1 p. 149-153 Open Access Repetitious DNA in mammalian chromosomes Albert de la Chapelle, Corresponding Author Albert de la ChapelleFolkhälsan Institute of Genetics Apollogatan 1 Helsingfors 10, FinlandSearch for more papers by this authorJim Schröder, Jim SchröderSearch for more papers by this authorRitva-Kajsa Selander, Ritva-Kajsa SelanderSearch for more papers by this author Albert de la Chapelle, Corresponding Author Albert de la ChapelleFolkhälsan Institute of Genetics Apollogatan 1 Helsingfors 10, FinlandSearch for more papers by this authorJim Schröder, Jim SchröderSearch for more papers by this authorRitva-Kajsa Selander, Ritva-Kajsa SelanderSearch for more papers by this author First published: December 1971 https://doi.org/10.1111/j.1601-5223.1971.tb02430.xCitations: 1AboutPDF ToolsExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Literature cited Arrighi, F. E. and Hsu, T. C. 1971. Localization of heterochromatin in human chromosomes. Cytogenetics 10: 81– 86. Arrighi, F. E., Hsu, T. C., Saunders, P. and Saunders, G. F. 1970. Localization of repetitive DNA in the chromosomes of Microtus agrestis by means of in situ hybridization. Chromosoma 32: 224– 236. Caspersson, T., Lomakka, G. and Zech, L. 1971. The 24 fluorescence patterns of the human metaphase chromosomes — distinguishing characters and variability. Hereditas 67: 89– 102. Caspersson, T., Zech, L., Johansson, C. and Modest, E. J. 1970. Identification of human chromosomes by DNA-binding fluorescent agents. Chromosoma 30: 215– 227. Gagné, R., Tanguay, R. and Laberge, C. 1971. Differential staining patterns of heterochromatin in man. Nature 232: 29– 30. Hungerford, D. A. 1965. Leukocytes cultured from small inocula of whole blood and preparation of metaphase chromosomes by treatment with hypotonic KCl. Stain Technol. 40: 333– 338. Jones, K. W. 1970. Chromosomal and nuclear location of mouse satellite DNA in individual cells. Nature 225: 912– 919. Human chromosomes in the spotlight. 1971. — Nature 232: 1– 2. Pardue, M. L. and Gall, J. G. 1970. Chromosoma localization of mouse satellite DNA. Science 168: 1356– 1358. Riegler, R. 1966. Microfluorometric characterization of intracellular nucleic acids and nucleoproteins by acridine orange. Acta Physiol. Scand. 67 (Suppl. 267): 1– 121. Schmid, W. 1967. Heterochromatin in mammals. Arch. Julius Klaus — Stift. Vererbungsforsch. 42: 1– 60. Schnedl, W. 1971. Unterschiedliche Fluorescenz der beiden homologen Chromosomen Nr. 3 beim Menschen. Humangenetik 12: 59– 63. Schröder, J. and de la Chapelle, A. 1971. Fetal lymphocytes in the maternal blood. Blood (in press). Sumner, A. T., Evans, H. J. and Buckland, R. A. 1971. New technique for distinguishing between human chromosomes. Nature 232: 31– 32. Citing Literature Volume69, Issue1December 1971Pages 149-153 ReferencesRelatedInformation

Novel Histochemical Stain for Tinctorial Detection of Cancer and Neoplastic Cells

Zetiq has introduced a histochemical stain that claims to tinctorially identify cancer and neoplastic cells. Because of the potential importance of such a capability, we undertook to investigate Zetiq's CellDetect® staining technology as applied to cultured cell lines as well as an initial sample of clinical cases. This goal was pursued by investigating four types of comparisons: 1) cancer cell lines before and after differentiation; 2) cervical squamous-cell carcinoma (SCC) biopsies to non-neoplastic squamous epithelium; 3) SCCs to neoplastic, nonmalignant squamous epithelium; and 4) neo-plastic squamous cells to non-neoplastic squamous cells in cytological preparations. The clinical material was also stained with hematoxylin and eosin (biopsies) or Pap (cytologies) for diagnostic purposes. We found that all CellDetect®-stained cells exhibited one of the two tinctorial outcomes. Cell lines with malignant phenotype uniformly had red/purple cytoplasm, whereas the differentiated phenotype changed the color to blue/green. Moreover, these two color values are sufficiently distinct that they can be readily distinguished quantitatively with an ELISA reader when applied to cells in tissue culture. Biopsies of SCC and non-neoplastic tissues exhibit the same two color values: cancers had red/purple cytoplasm, whereas non-neoplastic tissues were green/blue stained. In contrast, neoplastic, nonmalignant tissues (dysplasias) stained red/purple, similar to SCCs. Cytological preparations gave similar results: neoplastic cells stained red/purple, whereas non-neoplastic cells exhibited green/blue cytoplasm. The study demonstrated that the staining was rapid and reproducible. Conclusion: The CellDetect® stain allows detecting cancer and neoplastic cells tinctorially based on a rapid, reproducible histochemical process.

Investigation and application progress of urine formed elements staining technology

The urine formed elements staining is of importance for diagnosis and differential diagnosis of urine system diseases. In this article, we describe several kinds of improved staining techniques, such as cytochemistry staining, fluorescent staining and textile dye staining. The prospect of urine formed elements staining technology is also described. Key words: Urinalysis; Staining and labeling

A color discriminating broad range cell staining technology for early detection of cell transformation

Background:Advanced diagnostic tools stand today at the heart of successful cancer treatment. CellDetect® is a new histochemical staining technology that enables color discrimination between normal cells and a wide variety of neoplastic tissues. Using this technology, normal cells are colored blue/green, while neoplastic cells color red. This tinctorial difference coincides with clear morphological visualization properties, mainly in tissue samples. Here we show that the CellDetect® technology can be deployed to distinguish normal cells from transformed cells and most significantly detect cells in their early pre-cancerous transformed state.Materials and Methods:In tissue culture, we studied the ability of the CellDetect® technology to color discriminate foci in a number of two stage transformation systems as well as in a well defined cellular model for cervical cancer development, using HPV16 transformed keratinocytes.Results:In all these cellular systems, the CellDetect® technology was able to sensitively show that all transformed cells, including pre-cancerous HPV 16 transformed cells, are colored red, whereas normal cells are colored blue/green. The staining technology was able to pick up: (i) early transformation events in the form of small type 1 foci (non-invasive, not piled up small, with parallel alignment of cells), and (ii) early HPV16 transformed cells, even prior to their ability to form colonies in soft agar. The study shows the utility of the CellDetect® technology in early detection of transformation events.