Luxol Fast Blue MBS Research Articles

Certification procedures used by the Biological Stain Commission for eriochrome cyanine R (C.I. 43820, Mordant blue 3)

ABSTRACTEriochrome cyanine R (C.I. 43820, Mordant blue 3), also known as chromoxane cyanine R and solochrome cyanine R, has been used as a biological stain since 1957. In conjunction with ferric ions, it provides selective blue coloration of the nuclei of cells in methods procedurally similar to commonly used progressive or regressive hemalum (aluminum-hematoxylin) stains. Eriochrome cyanine R also is used to stain the myelin sheaths of axons in nerve tissue; the results are visually similar to those in sections stained with luxol fast blue MBS (C.I. 74180, solvent blue 38) with selective blue coloration of myelin and erythrocytes. Eriochrome cyanine R is an article of commerce with many uses in industrial coloration and analytical chemistry; it can be used instead of either hematoxylin or luxol fast blue MBS, especially in the event of a shortage of either of the latter compounds. The Biological Stain Commission (BSC) will certify batches of eriochrome cyanine R that meet the criteria set out in this document. The criteria include satisfactory UV/visible spectra at pH 4 and pH 12 − 13, a dye content not less than 40% and not greater than 52% (calculated as the color acid; equivalent to 46 − 59% of the trisodium salt), and satisfactory performance in three staining methods: regressive for nuclei, progressive for nuclei and regressive for myelin.

Myelin-targeted, texaphyrin-based multimodal imaging agent for magnetic resonance and optical imaging.

Reliable methods of imaging myelin are essential to investigate the causes of demyelination and to study drugs that promote remyelination. Myelin-specific compounds can be developed into imaging probes to detect myelin with various imaging techniques. The development of multimodal myelin-specific imaging probes enables the use of orthogonal imaging techniques to accurately visualize myelin content and validate experimental results. Here, we describe the synthesis and application of multimodal myelin-specific imaging agents for light microscopy and magnetic resonance imaging. The imaging agents were synthesized by incorporating the structural features of luxol fast blue MBS, a myelin-specific histological stain, into texaphyrins coordinated to GdIII . These new complexes demonstrated absorption of visible light, emission of near-IR light, and relaxivity values greater than clinically approved contrast agents for magnetic resonance imaging. These properties enable the use of optical imaging and magnetic resonance imaging for visualization of myelin. We performed section- and en block-staining of ex vivo mouse brains to investigate the specificity for myelin of the new compounds. Images obtained from light microscopy and magnetic resonance imaging demonstrate that our complexes are retained in white matter structures and enable detection of myelin. Copyright © 2016 John Wiley & Sons, Ltd.

Dark and light neurons in the human brain, with special reference to their reactions to Golgi's silver nitrate, luxol fast blue MBS and azocarmine G.

Sections from the human somatosensory cortex were observed with a light microscope. The neurons were classified into light and dark ones. The light neurons were slightly stained with thionin, luxol fast blue MBS and azocarmine G (80% of all neurons). The dark neurons were more or less shrunken, and stained intensely with these dyes (20% of all neurons). Double staining with luxol fast blue MBS and azocarmine G was especially useful to demonstrate the dark neurons, since it clearly stained even their fine processes. Neither light nor dark neurons were reactive to nick end labeling for detection of DNA fragmentation. Triple staining with Golgi's silver nitrate (or Gallyas's ammoniacal silver carbonate), luxol fast blue MBS and azocarmine G show that the majority of the dark neurons were argyrophilic (argyrophilic dark neurons, 15% of all neurons), while some of them were not argyrophilic (non-argyrophilic dark neurons, 5% of all neurons). Triple staining also showed that the light neurons were only occasionally argyrophilic (argyrophilic light neurons, 5% of all neurons); usually, the light neurons were not argyrophilic (non-argyrophilic light neurons, 75% of all neurons). The results confirm that dark neurons usually represent certain populations of neurons in the human brain, and that they are basically identical to the argyrophilic neurons. The discussion suggests that the argyrophilic light and dark neurons are excited cells, the non-argyrophilic dark neurons are exhausted cells, and the non-argyrophilic light neurons are resting cells. Triple staining further demonstrated that some glial cells were darkened and stained with Golgi's silver nitrate or Gallyas's silver carbonate. Additional Golgi's silver block staining showed that the argyrophilic neurons stained by the conventional block staining method usually possessed a shrunken cell body, which was characteristic of the dark neurons.

A histoquantitative study of the striate cortex and lateral geniculate body in normal, blind and demented subjects.

The amount of myelin in the outer band of Baillarger in the human visual cortex stained by Luxol Fast Blue MBS, has been measured in normal individuals of wide age range, in a group of blind and severely visually impaired but otherwise normal individuals and in a series of demented patients of the Alzheimer and multi-infarct type who were apparently visually normal. The amount of myelin is found to be significantly reduced in the blind and demented groups. Examination of the lateral geniculate body in the blind cases demonstrated slight reduction in neuron numbers and marked reduction in mean neuron diameter. Neuronal cytoplasmic ribonucleic acid, nucleolar volume and numbers of tetraploid glial nuclei in the blind or visually impaired cases were significantly reduced when compared with controls, indicating reduced function in these neurons. Similar results were obtained on examining this nucleus in Alzheimer and multi-infarct cases, suggesting that the visual pathways may be involved in these conditions. It is concluded that loss of myelin from the outer band of Baillarger occurs in association with reduced function of neurons in the lateral geniculate body.

Quantitative histochemistry of myelin using Luxol Fast Blue MBS.

The amount of Luxol Fast Blue MBS in the band of Genarri was measured with two types of scanning microdensitometer and the optical density determined. The amount of stain measured was proportional to the section thickness employed, thus demonstrating that the dye has stoichiometric properties in tissue sections. Blocks of tissue treated with phospholipid solvents showed an increased uptake of stain, suggesting that phospholipids are not a primary substrate for the dye in myelin staining. The dye may, therefore, be used to quantify myelin in tissue sections.

Luxol fast blue MBS and phloxine; a stain for mitochondria.

The applicability of Luxol fast blue MBS as a 0.1% solution in 0.05% acetic acid to the staining of mitochondria, first recognized in rat kidney by Shanklin and Nassar (Stain Techn., 34: 257-60. 1959), was confirmed in various organs (formalin-Zenker and Regaud's fixations; paraffin embedding) of the mouse and bullfrog. In liver cells and in the epithelium of renal tubules, mitochondria were stained green, selectively and clearly. The dark cells of the renal tubules and the middle piece of sperms in both animals were conspicuously demonstrated by their dense assemblages of green granules. The periodic acid-Schiff procedure proposed by Shanklin and Nassar as a counterstain was replaced by staining in 0.5% aqueous phloxine, 2-3 min; differentiation in 5% phosphotungstic acid, 2 min; and washing in water, 5 min. This simplified and accelerated the techique, and gave a better color contrast. Advantages of Luxol fast blue MBS and phloxine staining over traditional methods for mitochondria in paraffin sections ...

Iron hematoxylin chelates. II. Histochemistry of myelin sheath stains.

Aldehyde blockage, methylation, acetylation, amine blockage, and 4 min, 30 min and 24 hr deamination of paraffin sections of cat spinal cord were followed by staining with Lendrum's phloxine-tartrazine, Luxol fast blue MBS, PAS, phosphotungstic acid-hematoxylin, and Weil stains. Only the effects on staining of myelin are reported. These histochemical procedures separated the 5 stains into 3 groups: (1) phloxine-tartrazine and Luxol fast blue, (2) PAS, and (3) Weil and phosphotungstic acid-hematoxylin. The 3 patterns indicate that these stains may attach to 3 different reactive molecular sites in the sections. For the Weil and phosphotungstic acid-hematoxylin, such reactive sites are probably a secondary or tertiary amine or both.

MORPHOLOGICAL AND CYTOCHEMICAL PROPERTIES OF THE HOLOCRINE CELLS IN THE EPIDIDYMIS OF THE MOUSE.

Holocrine secretory cells have been identified in the epithelium of the epididymal canal of the mouse. These cells develop from basal cells. During their differentiation they grow toward the lumen of the epididymal canal and come to form club-shaped structures with an expanded apical portion, a central nucleus and a thin stalk-like connection to the basement membrane. Mature holocrine cells are characterized by their high acid phosphatase and aliesterase activity. They also are highly active for succinic dehydrogenase, nicotinamide adenine dinucleotide diaphorase, and nicotinamide adenine dinucleotide phosphate diaphorase. Nucleoside diphosphatase, thiamine pyrophosphatase, adenosine triphosphatase, and alkaline nucleoside phosphatase are also found in these cells. These cells are also characterized by their reactivity with the Aoyama and periodic acid-Schiff reactions. They react moderately with the molybdate and Luxol Fast Blue MBS reactions for choline containing compounds. Mature holocrine cells may disintegrate in situ or may be discharged in toto into the lumen of the epididymal canal. Glycerylphosphorylcholine was identified in extracts prepared from sperm-free epididymides of mice. Glycerylphosphorylcholine reacts with Aoyama and periodic acid-Schiff reactions as do mature holocrine cells. This fact coupled with the identification of choline containing material in holocrine cells suggests that they may be one site for the formation of glycerylphosphorylcholine.

Luxol fast blue ARN: a new solvent azo dye with improved staining qualities for myelin and phospholipids.

Luxol fast blue ARN (Du Pont, C.I. solvent blue 37) is a diarylguanidine salt of a sulfonated azo dye. This dye was compared with other Luxol blue and Luxol black dyes. Luxol fast blue ARN has improved staining qualities for phospholipids and myelin, and can advantageously be substituted for Luxol fast blue MBS (MBSN). Appropriate staining times for a 0.1% dye solution in 95% ethanol (containing 0.02% acetic add) at 35°-40° C range from 2-3 hr. After staining, the sections should be rinsed in 95% ethanol, rinsed in distilled water, and differentiated for 2 sec in 0.005% Li2CO3, rinsed in 70% ethanol, washed in water, and counterstained as required. Phospholipids and myelin selectively stain deep blue. A fixative containing CaCl2, 1%; cetyltrimethylammonium bromide, 0.5%; and formaldehyde, 10%, in water gave excellent results with brain. However, 10% formalin can be used. The staining of the phospholipids is probably due to the formation of dye-phospholipid complexes.

Luxol fast blue as a stain for Mallory's alcoholic hyaline.

Luxol fast blue MBS (du Pont), which has frequently been used as a stain for phospholipids, stains Mallory's “alcoholic” hyaline a deep purplish blue. The stain is stable and provides histological appearances far superior to other methods. It is used on paraffin sections of tissue fixed in formalin or formalin-sublimate as a 0.1% solution in 90% alcohol at 60°C for 8 hr. Differentiation is made with 0.05% Li2CO3 and a red counterstain applied.

Effects of Inorganic Oxides, Sulfides, Chlorides, and Acid Chlorides on Metachromasia and Other Staining Properties

The ability of 17 inorganic compounds (POCl3, PSC13, PC13, P2O5, P2S5, P4S3, P4S7, PC15, Sb2O5, As2O5, BiOC12, SeOC12, SO2C12, Sb2S5, VOC12, SiC14 and CrO2Cl2) dissolved in pyridine or 2,2,4-trimethyl pentane, to enhance subsequent staining of tissue components with toluidine blue, phosphotungstic acid-hematoxylin (PTAH), leukofuchsin, and dihydroxydinaphthyl-disulfide (DDD) was studied. Eight of these compounds were also tested for ability to enhance staining with Alcian blue 8GN and Luxol fast blue MBS. Nine of the 17 compounds produced increased staining of certain tissue components with leukofuchsin, 13 with toluidine blue, 16 with PTAH, and 16 with DDD. The results suggest additional approaches to identification of tissue entities by induced metachromatic basophilia and leukofuchsin positivity as well as by the other stains studied, and also suggest a number of hitherto unstudied modes of reaction between the dyes used and reactive groups of tissue components. Many reactions of the compounds tested, ...

Luxol Fast Blue Mbs: A Stain for Phase Contrast Microscopy

A staining method to increase the contrast of sectioned material for phase contrast microscopy is described. Two stock solutions of the stain are required. The first is made by dissolving 2 gm of luxol fast blue MBS in 100 ml of 95% ethanol. The second solution is made up of 4 ml of a 29% aqueous solution of FeCl3, 95 ml of 95% ethanol, and 1 ml of concentrated HCl. The staining solution is made by mixing equal parts of the two solutions. Sections are deparaffinized and taken to 70% alcohol, stained for 1.5 hr, dehydrated, cleared and covered as usual.