Janus green B 

Janus green B is a supravital stain. Janus green B staining reaction is oxygen dependent, and is reversibly inhibited by cyanide. Janus green B has been used for staining peripheral nerves in live insects, lymphatic vessels of rabbits and mitochondria^[1][2][3].

Observing New Linear Structures inside Rabbit Lymphatic Vessels using Janus Green B Staining Method^[1]
(1) Preparation of experimental animals: Anesthesia is administered to 12-week-old New Zealand rabbits, and an incision is made on the left side or in the middle of the rabbit's abdomen to expose the large lymphatic vessel located next to the inferior vena cava.
(2) Preparation of Janus Green B staining solution: The dye concentration is 65%, meaning 1 gram of Janus Green B is dissolved in 100 milliliters of 99.9% ethanol, and it is preheated to 40°C.
(3) Staining: Using a 30.5-gauge needle, approximately 0.3 milliliters of the preheated Janus Green B staining solution is injected as slowly as possible into a lymph node or lymphatic vessel close to the inferior vena cava. The injection needle is gently withdrawn, and the flow of the Janus Green B staining solution in the lymphatic vessel is observed presenting a blue color.
(4) Washing and maintaining temperature: A preheated phosphate-buffered saline (pH 7.4) at 40°C is injected to remove any backflow of the Janus Green B staining solution and to maintain the tissue's temperature and active state.
(5) Observation and localization: The internal condition of the lymphatic vessels is carefully observed using a stereomicroscope to determine the location of well-stained areas. The focus of observation is adjusted according to the staining effect and individual differences.
(6) Fixing samples: The collected samples are immersed in a 10% neutral formalin solution for fixation as soon as possible.
(7) Additional staining (optional): A portion of the fixed samples is selected for secondary staining using a 0.1% (w/v) acridine orange PBS solution.
(8) Imaging: Confocal laser scanning microscopy is used for imaging analysis of the stained samples.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

511.06

C₃₀H₃₁ClN₆

2869-83-2

Solid

Brown to black

CN(C1=CC=C(/N=N/C2=CC3=[N+](C4=CC=CC=C4)C5=C(C=CC(N(CC)CC)=C5)N=C3C=C2)C=C1)C.[Cl-]

Room temperature in continental US; may vary elsewhere.

4°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Lee BC, et al. Novel threadlike structures (Bonghan ducts) inside lymphatic vessels of rabbits visualized with a Janus Green B staining method. Anat Rec B New Anat. 2005 Sep;286(1):1-7.  [Content Brief]

  [2]. LAZAROW A, et al. Studies on the enzymatic basis for the Janus green B staining reaction. J Histochem Cytochem. 1953 Jul;1(4):234-41.  [Content Brief]

  [3]. Yack JE. Janus Green B as a rapid, vital stain for peripheral nerves and chordotonal organs in insects. J Neurosci Methods. 1993 Aug;49(1-2):17-22.  [Content Brief]