DOCSLIB.ORG

Nile Blue Inducible Fluorescence of Tumour Cells

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Nile Blue Inducible Fluorescence of Tumour Cells INDUCIBLE FLUORESCENCE OF TUMOUR CELLS 1395 Nile Blue Inducible Fluorescence of Tumour Cells A. L. BASTOS and DANTE MARQUES Servio de Patologia Celular Calouste Gulbenkian, Instituto Portugues de Oncoldgia Francisco Centil, Lisboa 4, Portugal (Z. Naturforsdi. 27 b, 1395—1398 [1972] ; received June 6, 1972) Inducible fluorescence, tumor cells An oxazine dye, Nile Blue sulfate, induces a fluorescent reaction in cytoplasmic granules (NBIFG) of living or fixed tumour cells in the same manner as reported before for Thiazine dyes. The NBIFG correspond to phase contrast positive granules when cells are viewed by phase contrast microscopy. Fluorescence disappears from NBIFG in a matter of 2 —4 days and the bodies turn deep blue. These granules have a succinate dehydrogenase (SDG) activity and are negative for acid phosphatase, peroxidasic activity and porphyrin. The cytological findings support the assumption that Nile Blue sulfate forms a salt linkage with unsaturated fatty acids of NBIFG which also show an oxi-reductive activity. The molecular nature of the fluorophore (s) is (are) unknown. We have previously shown that thiazine dyes can Material and Methods induce a fluorescent reaction in cytoplasmic granu- les of tumour cells when freshly stained by these Tumour Cells dyes and examined by fluorescence microscopy 2. The staining procedures (supravital and in vivo 1 The fluorescent reaction is called primary when staining) were carried out using transplantable tumour cells of mice, Sarcoma 37 tumour cells and Ehrlich the granules are visualized immediately after the tumour cells growing in ascitic form. Cells were ob- staining; secondary, when the granules become tained from tumour-bearing mice 5 to 10 days after fluorescent only after exposure of the cells to light. cell transplantation. The experiments were carried out The primary reaction is observed only with a parti- while the tumour was maintained during 378 conse- cutive passages. cular batch of Toluidine Blue and the secondary reaction with all Thiazine dyes so far tested, in- Nile Blue Staining cluding samples obtained from several manufactu- A modification of L i 11 i e's3 technique for de- rers. monstration of fatty acids was used as follows: — The dye inducible fluorescent (DIF) granules are 1. Fix tumour cells in 4% formaldehyde saline at 4 C refractile, which makes their microscopic recogni- for 1 hours. tion easy (i. e. Interference microscopy); cytochemi- 2. Remove the excess fixative by cell centrifugation for cal reactions for lipids (Oil Red 0, Sudan Black 6-10 min at 1000 rpm. and reduction of osmium tetroxide) are positive. 3. Resuspend the cells in an aqueous solution of Nilft Blue (0.005% w/v) and stain for 30 min. Sudanophilia and osmiophilia usually indicate 4. Mount the cell sediment in glycerol. the presence of unsaturated fatty acids. Since the 5. Examine by fluorescence microscopy. presence of these highly oxidizable lipids could help to explain the cytochemical nature of the DIF Fixatives reaction, L i 11 i es Nile Blue procedure was used to To study the effect of fixation the following fluids demonstrate unsaturated fatty acids. were used: Neutral formalin at 10% and 15%, Car- noy's fluid, Alcohol-Ether, 2.5% glutaraldehyde in The observations now reported show that Nile cacodylate HCl buffer. Blue sulfate (samples from several manufacturers) also gave the DIF reaction in the same granules Control of pH, dye concentration, temperature and stained by Thiazine dyes, both in living and fixed staining time cells. a) pH: The cells were stained in Nile Blue solution at pH 0.9 -12. The pH was buffer adjusted with NaOH Requests for reprints should be sent to Dr. A. L. BASTOS, (saturated solution). Above pH 9 the colour of the Servio de Patologia Celular Calouste Gulbenkian, Instituto solution changes from blue to pink and precipitates. Portugues de Oncologia Francisco Gentil, Lisboa 4, Portu- b) Dye concentration and tempera- gal. ture: The dye concentration in the staining solution 1396 A. L. BASTOS AND D. MARQUES was varied in tenfold dilutions from 0.5% to 0.00005% perchloric acid-mercapto ethylamine UV method for (w/v). The ratio of the cell volume to the volume of porphyrins 7. staining solution was 1 : 10. The temperature of the The above cytochemical preparations were then staining solution ranged from 0 °C to 100 °C. stained by Nile Blue sulfate to demonstrate the pre- c) Staining time: The staining times used sence of DIF granules and slides examined both by were als follows: - 0', T, 5', 10', 15', 30', 60', 2 hrs., light and fluorescence microscopy. 12 hrs., 24 hrs., 48 hrs. Nile Blue Compounds tested Results Nile Blue Sulfate [C.I. (913)51180] British Drug Houses, Poole, England, Batch N. 2868090; Matheson The fluorescence microscopic appearance of vi- Coleman & Bell, Norwood, Ohio, USA, Batch N. tally and supravitally stained and fixed tumour cells 481208; Chroma-Gesellschaft Schmid & Co., Stuttgart, are shown in Figs. 1,2* and 3. The Nile Blue indu- Germany, Batch unknown. All samples gave the fluo- rescent reaction. y Microscopic equipment _,40 UJ A Wild M20 microscope with a dual illumination O CC system for fluorescence, bright field and phase contrast UJ was used. It was fitted with Xenon lamp XBO 150w CL for fluorescence and a 12V/l00w quartziodine lamp for bright field and phase contrast. O <Z CL Fluorescence CD For Ultraviolet excitation — (c. 365 nm) the filter set consisted of a heat-absorbing filter KG 1 2, UG 1 3 4 O excitation filter . Red absorbing filter BG 38 and O 1 i GG 13 as the barrier filter. For Blue light excita- UJ tion — (c. 400 nm) a BG 12 3 excitation filter and C<O OG 1 3 barrier filter were used. £10 U. Cytochemistry O ca UJ In order to investigate the biological nature of DIF m granules, we employed the GOMORI lead phosphate ZD Z method 4 for lysosomes, peroxidactic activity in peroxi- 0 10 20 30 40 x somes, using 3 — 3' diamino benzidine (DBA) at pH 9 6, NUMBER OF FLUORESCENT GRANULES PER CELL succinodehydrogenase activity in mitochondria using a Graph I. Correlation of phase positive granules to fluorescent non-lipid soluble formazan forming salt-Nitro-BT 6 and bodies in 50 tumour cells. Fig. 1. Fluorescence microphotograph of ascitic Sarcoma 37. Figs. 6 a and 6 b. Fluorescence microphotograph of formalin Tumour cells vitally stained by Nile Blue. Bright green (ultra- fixed Nile Blue stained cells. Fluorescent emission disappears violet excitation) granules occur in the cytoplasm of almost in two days becoming the granules deep blue in bright field, every cell. Nucleus are non-fluorescent. 800 x. Fig. 6 b. 1200 x. Figs. 7 a and 7 b. Fluorescence microphotograph of an acid Fig. 2. Sarcoma 37 cells supravitally stained by Nile Blue. phosphatase preparation of Nile Blue stained tumour cells. Green colour emission of cytoplasmic granules as in vital The NBIFG do not show acid phosphatase activity although staining. 800 x. there are at least eight darker stained cells showing acid phos- phatase in bright field, Fig. 7 b. 1200 x. Fig. 4 a. Formalin fixed Sarcoma 37 tumour cell stained by Figs. 8 a and 8 b. Fluorescence microphotograph of peroxidac- Nile Blue and observed under bright field. No cytoplasmic tic activity in Sarcoma 37 tumour cells stained by Nile Blue. granules are seen. 2000 x. The NBIFG show no activity when the erythrocytes and neu- trophil have a positive reaction in the same preparation, Fig. 4 b. Phase contrast microphotograph of the cell shown Fig. 8 b. 1200 x. in Fig. 4 a. Note the presence of more than ten phase contrast Figs. 9 a and 9 b. Fluorescence microphotograph of five Sar- positive granules and four macro nucleolus. coma 37 tumour cells stained for succinic dehydrogenase and DIF reaction. Note the distribution of succinic dehydrogenase Fig. 5 a. Phase contrast microphotograph of formalin fixed activity in the same cells photographed in bright field, Fig. Nile Blue stained Sarcoma 37 tumour cells showing the pre- 9 b. 1200 x. sence of cytoplasmic granules. 1200 x. Figs. 10 a and 10 b. Diffuse cytoplasmic porphyrin emission Fig. 5 b. Fluorescence microphotograph of the same field as (red) of tumour cells which is weak when compared to the Fig. 5 a. Note the size, form and distribution of fluorescent reaction in erythrocytes (arrows). When these porphyrin pre- granules. parations of tumour cells are stained by Nile Blue (10 b) DIF granules are present showing yellow emission with blue light * Fig. 2 s. Tafel S. 1396 a. excitation. 1200 x. A. L. BASTOS and D. MARQUES. Nile Blue Inducible Fluorescence of Tumour Cells (S. 1395) Zeitschrift für Naturforschung 27 b, Seitel396 a Zeitschrift für Naturforschung 27 b. Seite 1396 b INDUCIBLE FLUORESCENCE OF TUMOUR CELLS 1397 V fluorescence microscopy. Graph 1 represents the cor- (* relation of phase-positive granules per cell to * * e fluorescent granules per cell. * A constant observation of Nile Blue stained O „ (pH 5) fixed cells is the disappearance of the # fluorescence in a matter of 2 — 4 days with the gra- * •' ' * * nules turning deep blue. § % Visual control of the preparations obtained i- > 0 under different staining conditions (Dye concentra- * * tion, temperature and staining time) showed that t fluorescence of NBIF granules (Excitation at 400) » was not affected by these variables. Figs. 7 to 10 illustrate simultaneously acid phos- • *\ phatase, peroxidactic activity, succinodehydro- > » genase, porphyrin and NBIF bodies in the same Fig. 3. Formalin fixed Sarcoma 37 tumour cells stained by preparations. These NBIF granules exhibit an asso- Nile Blue. Bright yellow emission of NBIFG with blue light ciated SDG activity (Figs. 9 a —9 b) with demon- excitation. 800 x. strable fluorescence in all cytochemical double cible fluorescent (NBIF) granules are identical to stained preparations.
Load more

Recommended publications
  • New Nile Blue Derivatives As NIR Fluorescent Probes and Antifungal Agents †
    Proceedings New Nile Blue Derivatives as NIR Fluorescent Probes † and Antifungal Agents Rui P. C. L. Sousa 1,2, João C. C. Ferreira 1,2, Maria João M. F. Sousa 2 and M. Sameiro T. Gonçalves 1,* 1 Centre of Chemistry, University of Minho, Campus of Gualtar, 4710-057 Braga, Portugal 2 Centre of Molecular and Environmental Biology, University of Minho, Campus of Gualtar, 4710-057 Braga, Portugal * Correspondence: [email protected] † Presented at the 22nd International Electronic Conference on Synthetic Organic Chemistry, 15 November– 15 December 2018; Available Online: https://sciforum.net/conference/ecsoc-22. Published: 14 November 2018 Abstract: The synthesis of four new Nile Blue derivatives with hydrogen, propyl and/or aminopropyl groups as substituents of the amines of 5- and 9-positions is described. Photophysical properties were evaluated in acidified ethanol and aqueous solution at physiological pH. Antifungal activity is also studied through the obtention of MIC values. Keywords: benzo[a]phenoxazines; Nile Blue derivatives; NIR fluorescent probes; antifungal agents 1. Introduction The development of new near-Infrared (NIR) fluorescent probes is a very important issue due to the wide range of applications [1–4]. These probes are an excellent choice to label biological material since its emission will not interfere with the natural fluorescence of biological compounds. Benzo[a]phenoxazinium salts, with Nile Blue being the best known, display fluorescence at around 600 nm and have been used as covalent and non-covalent fluorescent probes for amino acids, proteins and DNA, among other biological material [5–10]. In addition, applications as sensors or agents for photodynamic therapy (PDT) have been described [11,12].
    [Show full text]
  • Nile Blue Is a Basic Dye of the Oxazine Group, Which Has Been Used in Several Microscopic and Histochemical Techniques (4, 6)
    ACTA HISTOCHEM. CYTOCHEM. Vol. 16, No. 3, 1983 LETTER TO THE EDITOR NILE BLUE SULFATE STAINING FOR DEMONSTRATION OF LIPIDS IN FLUORESCENCE MICROSCOPY Nile blue is a basic dye of the oxazine group, which has been used in several microscopic and histochemical techniques (4, 6) . A staining method based on application of Nile blue sulfate was introduced early for the cytochemical demon- stration of lipids (3, 8, 9), showing neutral fats and fatty acids in red and blue, respectively. There is evidence (1, 10) that aqueous solutions of Nile blue sulfate contain the blue cation of the dye, a red oxidation product (Nile red), and the orange-red imino base. According to this view, phospholipids and fatty acids stain blue because they react with the Nile blue cation, meanwhile neutral fats appear in red color. Other observations seem to be in disagreement with this staining mecha- nism (5, 6) . A fluorescence reaction in tissues after staining with Nile blue sulfate and brilliant cresyl blue has been found by Bozzo and Campos Vidal (2). During the course of investigations by using oxazine dyes we have observed that Nile blue sulfate produces a strong fluorescence reaction in lipid droplets. The fat body of Drosophila larvae is a continuous tissue mass which has been chosen as test material because of its abundance in lipidic inclusions. Drosophila hydei testes, surrounded by the fat body, were fixed in 5 % formaldehyde for 1-24 hr, washed, and then immersed in 0.1 mg/ml aqueous Nile blue sulfate (Fluka) for 30 min. After staining, the material was briefly washed and mounted with a drop of water.
    [Show full text]
  • Dna Revealed
    Staining DNA on electrophoresis gels dna revealed Ethidium bromide, a potent mutagen Staining DNA on the move Concentrated In research laboratories, ethidium bromide and similar Recently, several commercial products have emerged that DNA Stain Dil e ute wi volum o th an equal f di use stilled water before fluorescent compounds are normally used to visualise DNA enable the DNA to be seen as it moves across the gel. Sto . re eze N at 4 °C. Do not fre AT N IO IO NA AT L C DUC ENTR GY E E FOR BIOTECHNOLO T HE U ING on a gel. Unfortunately, ethidium bromide and its breakdown Suppliers seldom reveal their composition, but several of NIVERSITY OF READ products are potent mutagens and carcinogens and therefore these stains contain Nile blue sulphate (also known as they should not be used in schools. Such dyes are often flat Nile blue A), a dye which had not previously been noted molecules with similar dimensions to DNA base pairs. When for its ability to stain DNA. Adkins and Burmeister (1996) ethidium bromide binds to DNA, it slips between adjacent give useful guidance as to its use as well as hints for identifying base pairs and stretches the double helix. This explains the other dyes which may be useful for visualising DNA. resources Methylene blue dye’s mutagenic effect — the ‘extra bases’ cause errors when the DNA replicates. In addition, short-wavelength UV light All of the dyes used for staining ‘mobile’ DNA are cationic Yung-Sharp, D. and (which itself is harmful) is required for ethidium bromide — that is, they are positively charged in the gel buffer, at Kumar, R.
    [Show full text]
  • Overcoming Cloning Problems by Staining Agarose Gels with Crystal Violet Instead of Ethidium Bromide in Lactate Dehydrogenase Ge
    Acta Biologica Hungarica 56 (3–4), pp. 389–397 (2005) OVERCOMING CLONING PROBLEMS BY STAINING AGAROSE GELS WITH CRYSTAL VIOLET INSTEAD OF ETHIDIUM BROMIDE IN LACTATE DEHYDROGENASE GENE FROM PLASMODIUM VIVAX AND PLASMODIUM FALCIPARUM D. TURGUT-BALIK,1* V. ÇELIK,1 KATH MORETON2 and R. L. BRADY2 1 Department of Biology, Faculty of Arts and Sciences, University of F°rat, Elaz°=, Turkey 2 Department of Biochemistry, University of Bristol, University Walk, Bristol BS8 1TD, U.K. (Received: August 9, 2004; accepted: November 9, 2004) In this study, lactate dehydrogenase gene from Plasmodium vivax has been tried to subclone into an expression vector. Some of the Plasmodium falciparum lactate dehydrogenase mutant genes have also been tried to clone and subclone into a vector, but we failed to clone or subclone either of the genes. DNA visualisation in electrophoretic gels typically requires UV radiation and the fluorecent dye ethidium bro- mide. A crystal violet-stained gel was run instead of an ethidium bromide gel and so avoided the use of UV radiation. This enabled us to clone or subclone both Plasmodium vivax lactate dehydrogenase gene and Plasmodium falciparum lactate dehydrogenase mutant genes into any desired vector. Keywords: Plasmodium vivax – Plasmodium falciparum – lactate dehydrogenase – crystal violet – ethid- ium bromide INTRODUCTION Drug resistance of Plasmodium to currently available antimalarials is increasing throughout the world. Discovery of a new, effective, safe, permanent and inexpen- sive antimalarial is essential to overcome this problem. The enzyme lactate dehydrogenase (LDH) has been targeted for the design of a novel antimalarial drug. It is a vital enzyme for the malarial parasite.
    [Show full text]
  • A Consideration of the Term Gloeocystidium
    April 1976 MEMOIRS OF THE NEW YORK BOTANICAL GARDEN 28(1): 123-130 A CONSIDERATION OF THE TERM GLOEOCYSTIDIUM MICHAEL J. LARSEN AND HAROLD H. BURDSALL, JR. Center for Forest Mycology Research, Forest Products Laboratory, Forest Service, U. S. Department of Agriculture, Madison, WI 53705 Structures termed “gloeocystidia” occur in diverse genera throughout the major groups of Homobasidiomycetes and have been variously defined: Ainsworth et al. (1971) state that it is a cystidium “that is thin-walled, usually irregular and with highly refractive hyaline or yellowish contents.” Snell and Dick ( 197 1) list the variant spelling “gleocystidium” with the definition, “A special form of cystidium in Hymenomycetes, of gelatinous or horny consistency and with oily, resinous, or granular contents.” Talbot (1954) and Price (1975) have provided comprehensive statements on the concept of gloeocystidia. We present a condensed version here of Talbot’s (1954, p. 288) definition. Sterile organs, with thin walls; lack of sculpturing and encrustation; contents hyaline to brownish, highly refractive, homogeneous, granular, or oily; aris­ ing from subhymenial and contextual tissues; staining deeply in phloxine and eosine in KOH mounts and becoming brown in iodine solutions. In 1944, Romagnesi proposed the term “macrocystide” for a cystidial form in the “Lactario-russulés,” and these cystidia (macrocystidia) were described as “trés longue . fusiform ou claviforme, souvent terminée par une pointe ou un appendice variable; son pédicule est très long et souvent en connexion avec laticifères de la trame,” and secondly “très souvent, mais non toujours, devient gris-bleu ou noirâtre . au contact de la sulfovanilline. .” Romagnesi’s ( 1944) interpretation of macrocystidia is apparently based primarily on form, and secondarily on the chemical reaction with sulfovanillin.
    [Show full text]
  • Analysis of Polyhydroxyalkanoates Granules in Haloferax Mediterranei by Double-Fluorescence Staining with Nile Red and SYBR Green by Confocal Fluorescence Microscopy
    polymers Article Analysis of Polyhydroxyalkanoates Granules in Haloferax mediterranei by Double-Fluorescence Staining with Nile Red and SYBR Green by Confocal Fluorescence Microscopy Verónica Cánovas 1,2,* , Salvador Garcia-Chumillas 1,2, Fuensanta Monzó 1, Lorena Simó-Cabrera 3,4 , Carmen Fernández-Ayuso 1, Carmen Pire 3,4 and Rosa María Martínez-Espinosa 3,4,* 1 Technological Centre of Footwear and Plastic of the Region of Murcia (CETEC) Avda, Europa 4-5, 30840 Alhama de Murcia, Spain; [email protected] (S.G.-C.); [email protected] (F.M.); [email protected] (C.F.-A.) 2 Cetec Biotechnology, Avda, Europa 4-5, 30840 Alhama de Murcia, Spain 3 Department of Agrochemistry and Biochemistry, Biochemistry and Molecular Biology Division, Faculty of Science, University of Alicante, Carretera San Vicente del Raspeig s/n, San Vicente del Raspeig, 03690 Alicante, Spain; [email protected] (L.S.-C.); [email protected] (C.P.) 4 Multidisciplinary Institute for Environmental Studies “Ramón Margalef”, University of Alicante, Ap. 99, 03080 Alicante, Spain * Correspondence: [email protected] (V.C.); [email protected] (R.M.M.-E.); Tel.: +34-968-662-200 (V.C.); +34-965-903-400 (ext. 1258) (R.M.M.-E.) Abstract: Haloferax mediterranei is a haloarchaeon of high interest in biotechnology because it produces Citation: Cánovas, V.; and mobilizes intracellular polyhydroxyalkanoate (PHA) granules during growth under stress Garcia-Chumillas, S.; Monzó, F.; conditions (limitation of phosphorous in the culture media), among other interesting metabolites Simó-Cabrera, L.; Fernández-Ayuso, (enzymes, carotenoids, etc.). The capability of PHA production by microbes can be monitored with C.; Pire, C.; Martínez-Espinosa, R.M.
    [Show full text]
  • Lysosomal Localization and Mechanism of Uptake of Nile Blue Photosensitizers in Tumor Cells1
    [CANCER RESEARCH 51, 2710-2719, May 15, 1991] Lysosomal Localization and Mechanism of Uptake of Nile Blue Photosensitizers in Tumor Cells1 Chi-Wei Lin,2 Janine R. Shulok, Sandra D. Kirley, Louis Cincotta, and James W. Foley Urology Research Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114 [C-W. L., J. R. S., S. D. KJ, and Rowland Institute for Science, Cambridge, Massachusetts 02142 ¡L,C., J. W. F.] ABSTRACT tosensitizers with high tumor selectivity will enable effective treatment of multiple, infiltratili!*, and invisible tumors, thus Nile blue derivatives have been shown to be potentially effective expanding the utility of PDT as a useful tool in cancer therapy photosensitizers for photodynamic therapy of malignant tumors. Results with intent to cure. Active research is under way to search for of a previous study suggested that the high accumulation of these dyes more tumor-selective sensitizers (4-6) and to improve the sen- in cells may be the result of dye aggregation, partition in membrane lipids, and/or sequestration in subcellular organelles. In this report, sitizer delivery system for better tumor targeting (7-10). results of studies are presented from an investigation of the subcellular Several early studies with animal tumor models have shown localization and mechanism of accumulation of these dyes in cells in that benzophenoxazines, including several Nile blue analogues, vitro. A video-enhanced fluorescence microscopy was used, and a punctate constitute a special class of dyes that are selectively localized in pattern of fluorescence was seen, most of which was localized in the tumors (11-15).
    [Show full text]
  • Handbook of Biological Dyes and Stains Synthesis and Industrial Applications
    HANDBOOK OF BIOLOGICAL DYES AND STAINS SYNTHESIS AND INDUSTRIAL APPLICATIONS R. W. SABNIS Pfizer Inc. Madison, NJ HANDBOOK OF BIOLOGICAL DYES AND STAINS HANDBOOK OF BIOLOGICAL DYES AND STAINS SYNTHESIS AND INDUSTRIAL APPLICATIONS R. W. SABNIS Pfizer Inc. Madison, NJ Copyright Ó 2010 by John Wiley & Sons, Inc. All rights reserved. Published by John Wiley & Sons, Inc., Hoboken, New Jersey Published simultaneously in Canada No part of this publication may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, scanning, or otherwise, exckpt as permitted under Section 107 or 108 of the 1976 United States Copyright Act, without either the prior written permission of the Publisher, or authorization though payment of the appropriate per-copy fee to the Copyright Clearance Center, Inc., 222 Rosewood Drive, Danvers, MA 01923, (978) 750-8400, fax (978) 750-4470, or on the web at www.copyright.com. Requests to the Publisher for permission should be addressed to the Permissions Department, John Wiley & Sons, Inc., 111 kver Street, Hoboken, NJ 07030, (201) 748-601 1, fax (201) 748-6008, or online at http://www.wiley.com/go/permission. Limit of Liability/Disclaimer of Warranty: While the publisher and author have used their best efforts in preparing this book, they make no representations or warranties with respect to the accuracy or completeness of the contents of this book and specifically disclaim any implied warranties of merchantability or fitness for a particular purpose. No warranty may be created or extended by sales representatives or written sales materials.
    [Show full text]
  • Supporting Information a Nile Blue Based Infrared Fluorescent Probe
    Electronic Supplementary Material (ESI) for ChemComm. This journal is © The Royal Society of Chemistry 2014 Supporting information A Nile Blue Based Infrared Fluorescent Probe: Imaging tumors that over-express Cyclooxygenase-2 Benhua Wanga, Jiangli Fan*a, Xianwu Wangb, Hao Zhua, Jingyun Wang*b, Huiying Mua, Xiaojun Penga a State Key Laboratory of Fine Chemicals, Dalian University of Technology, No. 2 Linggong Road, High-tech District, Dalian 116024, China. b Department School of Life Science and Biotechnology, Dalian University of Technology, No. 2 Linggong Road, Ganjingzi District, Dalian 116024, China. E-mail: [email protected], [email protected]. Content Materials and methods.....................................................................................................................S2 Synthesis of Compounds.................................................................................................................S2 Scheme S1. Synthetic procedures of Niblue-C6-IMC and its intermediates.................................S2 Photophysical properties of Niblue-C6-IMC.................................................................................S3 Cell incubation and staining with Niblue-C6-IMC........................................................................S4 Native-PAGE...................................................................................................................................S4 Fluorescence image of live cell stained with NBD C6-ceramide....................................................S4
    [Show full text]
  • Nile Blue 690 Perchlorate
    PO Box 31126 Dayton, OH 45437 Tel: 937.252.2989 Fax: 937.258.3937 E-mail: [email protected] www.exciton.com NILE BLUE 690 PERCHLORATE Synonym: 5-amino-9-(diethylamino)-benzo[a]phenoxazin-7-ium perchlorate Catalog No.: 06900 CAS No.: 53340-16-2 Chemical Formula: C20H20N3O.CIO4 MW: 417.85 Appearance: Dark green crystals Structure: + (H5C2)2N O NH2 ClO - N 4 Lasing Wavelength Max. Range Pump Source Concentration Abs Fl (nm) (nm) (nm) Solvent (molar) λ-max λ-max _________________________________________________________________________________________________ 705 FL3 Methanol 624e 660m 717 689-750 FL11a Methanol 628e 722 FL Methanol 678 656-719 XeCl(308)110 Methanol 7 x 10-4 710 696-740 XeCl(308)114 Methanol 2.3 x 10-3 698 680-723 XeF(351)154 Ethanol 8 x 10-4(NB690)+ 3.8 x 10-3(R610) 681 662-710 Nd:YAG(532)58 683 Nd:YAG→CV670(647)33 Methanol 2x10-4 109 -3 698 682-715 Nd:YAG(355) MeOH/H2O,3/2 1.5 x 10 (NB690), 1.5 x 10-4(R640) 111 -4 670 657-692 N2(337) Ethanol 4 x 10 (NB690), 1.9 x 10-3(R610) 114 -3 695 683-751 N2(337) Ethanol 1.1 x 10 5 -3 696 683-710 N2(337) Ethanol 3.8 x 10 (R610), 8 x 10-4(NB690) 90 729 688-783 N2(337) Ethanol 730 692-782 Kr(cw)14 EG 750 710-790 Kr(647)17 EG 1 x 10-3 695 682-730 Cu(511,578)175 Methanol 2 x 10-3 EG = Ethylene glycol; MeOH/H2O = Methanol/water; e = ethanol; m = methanol PO Box 31126 Dayton, OH 45437 Tel: 937.252.2989 Fax: 937.258.3937 E-mail: [email protected] www.exciton.com NILE BLUE 690 PERCHLORATE REFERENCES: 3.
    [Show full text]
  • Nile Blue-Based Nano-Sized Ph Sensors for Simultaneous Far- Red and Near-Infrared Live Bioimaging
    Nile blue-based nano-sized pH sensors for simultaneous far- red and near-infrared live bioimaging Jeppe Madsen, Irene Canton, Nicholas J. Warren, Efrosyni Themistou, Adam Blanazs, Burcin Ustbas, Xiaohe Tian, Russell Pearson, Giuseppe Battaglia, Andrew L. Lewis and Steven P. Armes* 1. Table of Contents 1. Table of Contents .......................................................................................................... 1 2. Tables ............................................................................................................................ 2 3. Figures ........................................................................................................................... 2 4. Materials ........................................................................................................................ 3 5. Synthesis of NBM monomer ......................................................................................... 4 6. Synthesis of NBC monomer .......................................................................................... 4 7. Reverse-phase HPLC analysis of Nile Blue monomers ................................................ 5 8. Dye purification ............................................................................................................. 5 9. Synthesis of PMPC macro-CTA via RAFT polymerization ......................................... 5 10. ATRP of PMPC with Nile Blue dye present throughout the polymerization ............ 6 11. ATRP of MPC with Nile Blue dye added at high MPC
    [Show full text]
  • Safety Data Sheet Packet
    Date of Issue: 23 March 2016 SAFETY DATA SHEET PACKET Product Identifier SRM Number: 1934 SRM Name: Fluorescent Dyes for Quantitative Flow Cytometry (Visible Spectral Range) SRM Description: This Standard Reference Material (SRM) is intended for use in assigning fluorescence intensity values to calibration standards for quantitative flow cytometry in the visible spectral range. A unit of SRM 1934 consists of four amber ampoules, each containing 2 mL of a different fluorophore solution or suspension. The solutions include Part A: Fluorescein Solution (60 μM aqueous borate buffer solution); Part B: Nile Red Solution (60 μM acetonitrile solution); Part C: Coumarin 30 Solution (acetonitrile solution); and Part D: APC Suspension (Allophycocyanin fluorescent protein in aqueous buffer solution, 100 µL). SRM 1934 Parts: Part A: Fluorescein Solution Part B: Nile Red Solution Part C: Coumarin 30 Solution Part D: APC Suspension Company Information National Institute of Standards and Technology Standard Reference Materials Program 100 Bureau Drive, Stop 2300 Gaithersburg, Maryland 20899-2300 Telephone: 301-975-2200 Emergency Telephone ChemTrec: FAX: 301-948-3730 1-800-424-9300 (North America) E-mail: [email protected] +1-703-527-3887 (International) Website: http://www.nist.gov/srm This page intentionally left blank. DATE: 23 March 2016 Product Identifier SRM Number: 1934 SRM Name: Fluorescent Dyes for Quantitative Flow Cytometry (Visible Spectral Range) SRM Part: Part A: Fluorescein Solution Under the U.S. Department of Labor, Occupational Safety and Health Administration (OSHA) 29 CFR 1910.1200, this Standard Reference Material (SRM) is NOT classified as a physical hazard or a health hazard, a simple asphyxiant, combustible dust, pyrophoric gas, or hazard not otherwise classified.
    [Show full text]