How to use a Spectrophotometer

Summary

This video tutorial demonstrates the operation of a spectrophotometer, a device used to measure the absorbance of various samples. It begins with the initialization process, which may take several minutes, followed by setting the desired wavelength, specifically 540 nm for this demonstration. Users are guided through blanking the machine with a solvent blank to eliminate solvent absorbance before inserting sample cuvettes for measurement. The tutorial emphasizes the importance of re-blanking when changing wavelengths and highlights the simplicity of measuring multiple samples consecutively without readjustment.

Takeaways

• 🔧 A spectrophotometer is used to measure the absorbance of various samples.
• 🔌 Before use, remove the cover and turn on the spectrophotometer.
• ⏳ The initialization process can take several minutes after turning on the device.
• 🌈 Set the desired wavelength using the 'NM arrow up' or 'NM arrow down' buttons.
• 📏 For this demonstration, the wavelength is set to 540 nm.
• 💧 To blank the machine, place a solvent blank in the chamber and press 'zero ABS 100% T'.
• 🆕 After blanking, the device displays the set wavelength and an absorbance reading of 0.0000.
• 📊 To read a sample, replace the solvent blank with the sample cuvette and close the chamber.
• 🧪 Absorbance readings for multiple samples can be taken without adjusting the wavelength again.
• 🔄 If the wavelength is changed, re-blank the machine following the same procedure.

Q & A

What is the primary function of a spectrophotometer?

A spectrophotometer is used to read the absorbance of various samples.

What should you do when you first receive a spectrophotometer?

You should remove the cover and turn it on.

What happens during the initialization process of the spectrophotometer?

The spectrophotometer goes through an initialization process that can take several minutes.

How do you set the wavelength on a spectrophotometer?

You set the wavelength using the buttons labeled NM arrow up or NM arrow down.

What solvent is required for blanking the machine?

A solvent blank is needed, which is prepared in a cuvette.

What does the message 'setting blank' indicate?

It indicates that the spectrophotometer is calibrating itself to ignore the absorbance due to the solvent.

What does the reading '0.0000 A' signify?

It signifies that the absorbance of the solvent blank has been zeroed out.

How do you read the absorbance of a sample?

You remove the solvent blank, insert the sample cuvette, close the chamber, and read the absorbance displayed.

What should you do if you want to read multiple samples at the same wavelength?

You can simply open the chamber, replace the cuvette with the next sample, close it, and read the absorbance without needing to adjust the wavelength.

When do you need to re-blank the machine?

You need to re-blank the machine any time you change the wavelength.