Protocol: Cleaning Glass Coverslips with Acid (1M HCl)

Glass coverslips are used for mounting specimens, and growing cells. Cleaning the coverslips with 1M HCl will remove dust particles and oily chemicals, thus enhancing their microscopic properties. Clean coverslips are also suitable for coating with poly-lysine and other types of coatings such as fibronectin, and laminin.


Reagents and solutions
1M Hydrochloric acid (HCl)
50% ethanol
Absolute ethanol (>99%)

Deionized/Double distilled water

Equipment and disposables
Measuring cylinder
Aluminium foil
Incubator (set between 50 – 60°C)
Orbital shaker

Acid washing of coverslips


Use appropriate personal protective equipment (lab coat, gloves, goggles, etc) for your safety and follow your institute’s guidelines.

Step 1: Take 100 ml 1 M hydrochloric acid (HCl) in a 250 ml beaker. Add one by one 50 – 100 coverslips to the beaker.

Make sure coverslips are not sticking to each other.

Step 2: Cover the beaker with aluminum foil tightly (to stop excessive evaporation of liquid) and incubate at 50 – 60°C with gentle shaking overnight.

Note: You need to add 473.68 ml water.

Step 3: Remove the HCl from the beaker and wash the coverslips with deionized water 4 – 5 times.
Add 150 – 200 ml deionized/double distilled water. Cover the beaker with fresh Aluminium foil and shake it on the orbital shaker for 15 min. Discard the water. repeat this 4 – 5 times.

Note: The pH of the water will be ≈7 after all washes are over.

Step 4: Discard the water from the beaker and add 100 ml 50% ethanol. Shake the beaker on the orbital shaker for 15 min. Discard the 50% ethanol.

Step 6: Add 100 ml of absolute ethanol (>99% ethanol). Shake the beaker on the orbital shaker for 15 min. Discard the ethanol.

Step 7: Dry the coverslips in a dust-free area.

Coverslips should be at room temperature in a tightly sealed container.

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