What can be used as a substitute for methylene blue?
You can substitute Methylene Blue in most staining protocols that call for carmine or Janus green B. Methylene blue is also used by aquarium hobbyists for preventing fungal disease. Wet Mounting Medium. A glycerol-based mounting medium can provide a high refractive index than pure water. It is necessary to see certain cellular structures.
How do you make methylene blue?
Methylene blue is a formal derivative of phenothiazine. It is a dark green powder that yields a blue solution in water.. How do you make methylene blue solution at home? Procedure: Dissolve 0.1g methylene blue in 100 ml distilled water (0.1% MB stock solution) • Mix 10 ml of the MB stock solution with 90 ml of the 0.1M glycine buffer. Do you need a prescription for methylene blue?
Why does methylene blue inhibit Gram positive bacteria?
The growth of Gram positive bacteria is generally inhibited on EMB agar because of the toxicity of methlyene blue dye.
Why is methylene blue or iodine necessary?
Why is methylene blue and iodine necessary? Methylene blue can be used in biology as a stain to indicate whether cells are dead or alive. When studying plant cells, iodine can stain not only starch, but also enters the cell wall-cell membrane pores and can facilitate the staining of the nucleus, rendering it more visible under the microscope.
What is the chemical name for methylene blue?
Chemical compound. New methylene blue (also NMB) is an organic compound of the thiazine class of heterocycles. It is used as a stain and as an antimicrobial agent. It is classified as an azine dye, and the chromophore is a cation, the anion is often unspecified.
What is NMB used for?
NMB is a staining agent used in diagnostic cytopathology and histopathology, typically for staining immature red blood cells. It is a supravital stain. It is closely related to methylene blue, an older stain in wide use.
How long to incubate methylene blue stain?
Mix equal amounts of blood and new methylene blue stain (2 to 3 drops, or approximately 50 μL each), and allow to incubate at room temperature for 3 to 10 minutes.16
What pH is blue in a stain?
Staining solution. 1.0% brilliant cresyl blue or New methylene blue in iso-osmotic phosphate buffer pH 7.4. New batches of stain must be tested with a known positive control because the redox action of the dyes may vary from batch to batch.
What color are haemoglobin H inclusions?
Patients with α thalassaemia, who form haemoglobin H (β 4), have red cells in which multiple blue-green spherical inclusions develop on exposure to brilliant cresyl blue or New methylene blue. This is mainly a feature of haemoglobin H disease, but small numbers of similar cells may be seen in α thalassaemia trait, particularly, but not only, in α0 thalassaemia heterozygosity. A modified method with a higher degree of specificity and a sensitivity of 94% has been proposed when haemoglobin H inclusions are used for the detection of α 0 thalassaemia. 58
What color is Heinz body?
Heinz bodies are also stained by New methylene blue, but they stain a lighter shade of blue than the reticulofilamentous material of reticulocytes and stain well with methyl violet ( Fig. 15-5 ).
What is the best blue for reticulocytes?
Better and more reliable results are obtained with New methylene blue than with brilliant cresyl blue. New methylene blue is chemically different from methylene blue, which is a poor reticulocyte stain. New methylene blue stains the reticulofilamentous material in reticulocytes more deeply and more uniformly than does brilliant cresyl blue, which varies from sample to sample in its staining ability. Azure B is a satisfactory substitute for New methylene blue; it has the advantage that the dye does not precipitate and it is available in pure form.41 It is used in the same concentration and the staining procedure is the same as with New methylene blue.
What color should reticulofilamentous blood be?
A larger proportion of anaemic blood, and a smaller proportion of polycythaemic blood, should be added than of normal blood. In a successful preparation, the reticulofilamentous material should be stained deep blue and the nonreticulated cells should be stained diffuse shades of pale greenish blue.
What color is Hb H?
In Hb H disease, α-thalassemia minor, and silent carrier α-thalassemia, brilliant cresyl blue or new methylene blue stain may be used to induce precipitation of the intrinsically unstable Hb H.43 Hb H inclusions (denatured β 4 tetramers) typically appear as small, multiple, irregularly shaped greenish-blue bodies that are uniformly distributed throughout the RBC. They produce a pitted pattern on the RBC surface similar to the pattern of a golf ball or raspberry ( Figure 25.7 ). In Hb H disease, almost all RBCs contain Hb H inclusions. 36 In α-thalassemia minor, only a few cells may contain these inclusions, and in silent carrier α-thalassemia, only a rare cell does. These inclusions appear different from Heinz bodies, which are larger and fewer in number and most often appear attached to the inner membrane of the RBC. This test is very sensitive in detecting Hb H in α-thalassemia syndromes. 43
Introduction
Methylene Blue Stain, also known as methylene blue aqueous is a ordinary stain. It is used for capsule stain as well as simple staining for a arpid and easy way to determine cell shape, size, and arrangements of bacteria.
Principle of Methylene Blue Stain
The capsule is distinct from the slime layer that most bacterial cells form in that it is a thick, detectable, discrete layer outside the cell wall. The capsule stain takes on an acidic stain while a basic stain to detect capsule production. A positive capsule stain requires a mordant that precipitates the capsule.
Preparation of Methylene Blue Stain
Weigh 0.5 gm methylene blue powder on a piece of clean paper (pre-weighed) and dissolve the stain in about 30 ml of water.
Staining Procedure of Methylene Blue Stain
Transfer aseptically a loopful of microbial culture on a clean and grease-free dry slide.
Limitations of Methylene Blue Stain
Capsules are more delicate since using heat during the staining process should be avoided given that capsules would be easily destroyed.
Keynotes on Methylene Blue Stain
Capsules are fragile and can be diminished, desiccated, distorted, or destroyed by heating.