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Trypan Blue Injection | Description, Uses and Side Effects

Home » Blog » Trypan Blue » Trypan Blue Injection | Description, Uses and Side Effects

In recent times, vital dyes and their use in ophthalmic surgery have multiplied in importance. Trypan blue Injection has been the best choice for anterior segment surgeons because of its safety, ease, and great ability to help surgery in hard situations that involve tissue visibility. Trypan blue has been put to use in cataract surgery for almost 10 years, and its use went on to include various anterior segment procedures like corneal transplantation and trabeculectomy. The techniques and outcomes of Trypan blue dye-assisted anterior segment surgeries will be discussed in this blog.

What is Trypan Blue Injection

Trypan Blue Injection is a sterile solution of Trypan Blue. Trypan blue has been the best method for staining dead cells to know the cell viability. This dye is not included from live cells that have intact membranes, although it can enter and focus in dead cells that have compromised membranes, making them dark blue. With no scientific support, there has been a belief that trypan blue is not accurate for cell viability below 80%. Trypan blue has been shown to change the morphology of cells that are dead to a diffuse shape, taking it to an overestimation of viability. When the cells were stained with propidium iodide, the same results were not seen—Trypan blue and propidium iodide co-localize, confirming that the diffuse objects are cells with nuclei. After the contact of trypan blue, the cell ruptures. The cells died outside the incubator, the formation of these diffuse objects was seen and counted over time. Rapid water influx may have made the cells rupture and vanish, as seen in the demonstration. Some dead cells have vanished after trypan blue staining. The total number of cells may be less, which results in an exaggeration of cell viability. This error can have an impact on the actual result of cellular therapies that rely on accurate measurements of the immune cells that will be infused back into patients.

Trypan blue Injection Brands

Brand Company Country
Blue Rhexis De Ceunynck Ophthalmology Belgium
Blue Rhexis (Trypan Blue Solution) Ophtho India Inc. India

Indications and Dosage

By the staining of the anterior capsule in the lens, trypan blue is indicated to use aid in ophthalmic surgery.

The dosage for cataract surgery:

A blunt cannula must be attached to a 2.25 mL syringe that contains trypan blue. After opening the eye, an air bubble is injected into the anterior chamber to stop trypan blue from being diluted by the aqueous. The blunt cannula is used to put trypan blue to the anterior lens capsule. As early as the dye gets in contact with the capsule, enough staining happens. To get all the excess dye, the anterior chamber is filled with a balanced salt solution. After that, an anterior capsulotomy can be performed.


15-25°C is the recommended temperature for storage of trypan blue. Keep it out of direct sunlight.

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Uses of Trypan blue Injection

In cataract surgery

In cataract surgery, trypan blue is used to stain the anterior capsule of the lens for capsulorhexis. As the vitreous capsule and Descemet’s membrane are basement membranes, Trypan blue bacteria improves Descemet’s membrane visibility during the cataract surgery.

In Identification of Retinal Breaks 

As per a research paper titled “Identification of Retinal Breaks Using Subretinal Trypan Blue Injection” trypan blue can be used to identify retinal breaks during vitrectomy for rhegmatogenous retinal detachment. For the purpose of the research Trypan blue 0.15% was injected transurethrally into the subretinal space using a 41-gauge cannula designed for macular translocation surgery.

Vital staining of the connective tissues

Trypan blue intravenously binds fast to portions of the intercellular connective tissue like the collagen, reticulin, and elastic fibres through the body. The previous observations of diffuse staining of the elastic fibres are simply a case of general affinity of connective tissue for dyes.

Side effects of Trypan blue injection

Discolouration of high water content hydrogen intraocular lenses and inadvertent staining of the posterior lens capsule and vitreous face has been known as the effects of using trypan blue. The posterior lens capsule staining of the vitreous face is often self-limiting and lasts up to one week.


Why is trypan blue used in cataract surgery?

Because of its safety and efficacy, trypan blue is mainly used in cataract surgery. Trypan blue has been known to be used since the year 1990 to colour the anterior capsule in cataract surgery.

How does trypan blue determine cell viability?

To know the number of viable cells in a cell suspension, the test of dye exclusion is used. They believe they live cells have intact cell membranes that keep trypan blue out, whereas dead cells don’t. 

Why do dead cells stain blue?

Dead cells lack these intact cell membranes, which can allow the dye to penetrate them and stain dead cells blue.


The information provided here is based on general knowledge, articles, research publications etc and we do not claim the authenticity of any of the information provided above. We do not claim or suggest/advise any medical, therapeutic, health or nutritional benefits of Trypan blue. We do not supply or promote our Trypan blue product for the applications which are covered by valid patents and which are not approved by the FDA.

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Trypan Blue | Chemistry, uses and side effects

Macsen Labs is a manufacturer and supplier of several grades of Trypan blue such as:-

Trypan Blue