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Gram staining kit

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Product Item Number
Product Specifications
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  • AS42L121
    100mL
    140
Product Description Product FAQ COA Product Literature MSDS

product description

Gram staining is a widely used identification staining method in bacteriology. The mordant iodine solution enters the bacterial cell and binds with crystal violet. Gram positive bacteria have a high and firm uptake of iodine and crystal violet, and are not easily decolorized by alcohol. Gram negative bacteria have low uptake of iodine and crystal violet, are unstable, and are easily decolorized by alcohol. For the convenience of observation, after decolorization, a red dye such as alkaline safranin is used for counterstaining. Positive bacteria remain purple, while negative bacteria are stained red.

ordering information

product name

Item number

specifications

Gram staining kit

AS42L121

100mL

Product components

components

specifications

A. Ammonium oxalate crystal violet dye solution

100mL

B. Gram iodine solution

100mL

C. Decolorization solution

100mL

B. Gram iodine solution

100mL

Transportation and storage

Transport at room temperature. Store at room temperature and avoid light, with a shelf life of 12 months.

Usage

1. Preparation

(1) Using a sterile inoculation ring, pick a thin layer of sample and place it on a glass slide for smear;

The inoculation ring is sterilized by burning with an alcohol lamp flame and can be cooled for 30 seconds before use;

If the tissue sample being tested may be contaminated with bacteria or if bacteria are growing in the culture medium, the sample can be directly picked up with an inoculation ring and placed on a glass slide. If detecting bacterial clones growing on a culture plate, add a drop of sterile water onto a glass slide, then pick up a clone and place it in water.

(2) Let the smear dry naturally, fix it with heat, and skip it 2-3 times on the flame of an alcohol lamp; Note: Do not keep the glass slide continuously on the flame to prevent the sample from overheating.

2. Staining

(1) Place the heat fixed glass slide in a dye vat containing ammonium oxalate crystal violet staining solution, and let the glass slide stand in the vat for 1 minute.

(2) Gently wash the glass slide with tap water for 5 seconds and shake off any excess water.

(3) Place the glass slide in a staining vat containing Gram iodine solution for 1 minute, wash with water, and shake off excess water.

3. Discoloration and re dyeing

(1) Tilt the glass slide slightly and add a decolorization droplet onto the top of the slide, allowing it to slide through the sample area until purple appears. It is important that decolorization should neither be excessive nor insufficient. If decolorization is excessive, Gram positive bacteria can be mistaken for negative bacteria; If the decolorization is insufficient, Gram negative bacteria can be misjudged as positive bacteria.

(2) Wash for 5 seconds and shake off excess water.

(3) Stain with safranin counterstain for 1 minute.

(4) Wash for 5 seconds and shake off excess water. Allow it to dry naturally, or carefully absorb the moisture with absorbent paper.

4. Microscopic examination: Observe bacteria under an oil microscope (900 ×~1000 ×) to distinguish between Gram positive and Gram negative bacteria. Do not use cover slips, add a drop of oil in the middle of the slide to observe bacteria. If necessary, the slides can be stored in a slide box for several months before observation.

Staining results: Gram positive bacteria stain blue purple, while Gram negative bacteria stain red.

matters needing attention

1. This product is only for scientific experimental research and should not be used in clinical diagnosis, treatment, or other fields.

2. For your safety and health, please wear lab clothes and disposable gloves when operating.

3. The key to Gram staining is to strictly control the degree of alcohol decolorization. If decolorization is excessive, positive bacteria can be mistakenly stained as negative bacteria; When decolorization is insufficient, negative bacteria can be mistakenly infected as positive bacteria. In addition, bacterial age also affects staining results, such as when positive bacteria are cultured for too long, or when they have died or some bacteria dissolve on their own, they often show negative reactions.

4. The specimen smear should not be too thick.

5. The temperature of the glass slide passing through the flame should not be too high.

6. If the smear is thick, extend the decolorization time until no more purple color appears.

7. If the iodine solution becomes transparent, it cannot be used.

8. When washing with water, gently move and rinse along the diagonal direction of the glass slide with a washing bottle to avoid washing away the bacterial cells.

9. When using this kit for the first time, it is recommended to take 1-2 samples for preliminary experiments.

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This product is only for scientific experimental research and should not be used in clinical diagnosis, treatment, or other fields.

Fill in the product batch number for querying

Fill in the product batch number for querying

Contact Information

Landline Phone Number:021-50778506 
Service Manager:18616108315

Address:4th Floor, Building 27, Lane 908, Ziping Road, Pudong New Area International Medical Park, Shanghai

E-mail:lsj0027@obiosh.com

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