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Key Documents

94448

Sigma-Aldrich

Gram′s crystal violet solution

for microscopy

Synonym(s):

Basic violet 3; gentian violet; hexamethyl pararosaniline; methyl violet 10B

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250 ML
€22.80
2.5 L
€107.00

€22.80

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250 ML
€22.80
2.5 L
€107.00

About This Item

UNSPSC Code:
12171500
NACRES:
MA.02

€22.80

Please contact Customer Service for Availability
Request a Bulk Order

grade

for microscopy

Quality Level

200

product line

BioChemika

form

liquid

shelf life

limited shelf life, expiry date on the label

technique(s)

microbe id | staining: suitable

color

purple to very dark purple

density

0.981 g/mL at 20 °C

εmax

33.8 at 585-595 nm in water

suitability

suitable for microscopy

antibiotic activity spectrum

Gram-negative bacteria
Gram-positive bacteria

application(s)

diagnostic assay manufacturing
hematology
histology

storage temp.

room temp

Related Categories

General description

Gram’s crystal violet solution, also known as gentian violet or hexamethyl pararosaniline, is a basic, cationic, acidotropic aminoarylmethane dye. It is a hexa-N-methylated triaminotriphenylmethane with a moderately sized lipophilic cation and a slightly nonplanar conjugated system. It is used in bacteriology as the primary stain in Gram staining to differentiate Gram-positive and Gram-negative bacteria. It is known to possess antibacterial and antifungal properties.[1][2]

Application

  • Crystal violet is routinely used for Gram staining and its variants, staining amyloid, bacterial components, and vascular plant tissues.
  • It is used in polychrome staining of epoxy resin sections, viability staining of cultured neurons, and confocal optical sectioning to analyze meiotic structures.
  • It is also employed in the acridine orange-crystal violet staining of intracellular bacteria, microsporidian spores, and cytological smears.

Biochem/physiol Actions

The antimicrobial effects of crystal violet are caused by free radical generation and the formation of unionized dye-bacteria complexes. It also inhibits bacterial protein synthesis and cell wall formation and is especially effective against Gram-positive bacteria.[2]

Principle

Crystal violet is the primary stain in Gram staining. It infiltrates the peptidoglycan-rich cell wall of Gram-positive bacteria, and forms complexes with Gram’s iodine that are entrapped and retain the purple color even after decolorization. Thus, Gram-positive bacteria can be differentiated from Gram-negative bacteria whose peptidoglycan layer is considerably thinner and does not retain crystal violet.

Signal Word

Warning

Hazard Statements

H226,H319,H351,H412

Hazard Classifications

Aquatic Chronic 3 - Carc. 2 - Eye Irrit. 2 - Flam. Liq. 3

Storage Class Code

3 - Flammable liquids

WGK

WGK 2

Flash Point(F)

93.2 °F

Flash Point(C)

34 °C

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Regulatory Listings

Regulatory Listings are mainly provided for chemical products. Only limited information can be provided here for non-chemical products. No entry means none of the components are listed. It is the user’s obligation to ensure the safe and legal use of the product.

EU REACH Annex XVII (Restriction List)

CAS No.

Choose from one of the most recent versions:

Certificates of Analysis (COA)

Lot/Batch Number
BCCM6957
BCCL8948
BCCK2162
BCCL0880
BCCJ8029

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If you require a particular version, you can look up a specific certificate by the Lot or Batch number.

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Alexander M Maley et al.
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Questions

1–3 of 3 Questions  

  1. If the concentration of a solution is 2% and a final concentration of 0.01%, should water or ethanol be added to further dilute it?

    1 answer

    1. Product 94448 is intended for use in a Gram Stain procedure. If you are diluting it down to 0.01%, you are not using the product as intended for staining bacteria in a Gram Stain procedure. If the concentration is a 0.01% solution, this brings attention towards the use of Turk's solution for counting cells using a hemacytometer. Turk's solution can be formulated with or without acetic acid. Using 10 to 20% ethanol or a higher concentration effectively kills the cells, causing the entire cell to stain and making it impossible to identify cells by the shape of their nuclei.

      If it is for a migration assay or some other use, it would be advisable to check the literature. In some instances, the standard procedure is to dilute the Crystal Violet in methanol. Crystal Violet has many and varied uses, so it is best to try different solvents and determine which is most appropriate for your use. A 2 percent solution can be diluted to a 0.1 percent solution with a 1:20 dilution in an appropriate solvent, such as water, 15% ethanol, or 100% methanol.

      Helpful?

  2. What is the concentration of Crystal Violet in 94448?

    1 answer

    1. The SDS states that the concentration of the Crystal Violet is between 1 and 2.5%. Crystal Violet in product HT901, listed as 2.3% in the Instructions For Use. The solubility of Crystal Violet in solution with ethanol present can exceed 1%, while in a solution with just water, solubilities are typically around 4 mg/ml or 0.4%.

      Helpful?

  3. What is the concentration of this product?

    1 answer

    1. The solution of Crystal violet-2g in 20ml of ethanol(95%) is added to 0.8g of Ammonium oxalate in 80 ml of deionized water. Therefore, the solution has a concentration of about 2%.

      Helpful?

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