Cytospin Instructional Video

Summary

This instructional guide walks users through the process of preparing microscope slides of cells using centrifugation. It covers the proper placement of slides in the cassette, the addition of filters and funnels, and the correct pipetting of cell samples. Key steps include balancing slides in the rotor, setting up the centrifuge, and ensuring the seal is secure. The video also highlights the importance of careful handling when removing slides to avoid damage to the cells. The entire process, ideal for eukaryotic cells, is completed in about 3 minutes at 800 rpm.

Takeaways

• 😀 The rotor must be vacuum sealed before opening to avoid damaging its spindle.
• 😀 To open the rotor, pull up on the central rod to release the seal, then remove the top.
• 😀 To reset the vacuum seal, ensure the plastic top is firmly in place and the center rod is pushed down.
• 😀 Check the seal by pulling on the blue handle before placing the rotor back on the spindle.
• 😀 Microscope slides must be specially prepared by placing them in a metal cassette with the frost side facing the hinge.
• 😀 Place a filter on top of the slide with an unused hole aligned with the corresponding hole in the cassette.
• 😀 Position the funnel on top of the filter, ensuring it opens into the hole, and clasp the clamp on the cassette.
• 😀 Make sure all parts of the preparation are aligned so no holes are covered by other parts.
• 😀 Pipette 100 microliters of the sample into the funnel and place the prepared slide into the rotor.
• 😀 Balance the centrifuge by ensuring each slide has a corresponding slide across from it in the rotor.
• 😀 Set the centrifuge to 800 rpm for 3 minutes to spin the eukaryotic cells, and listen for the jingle when it's done.
• 😀 After the centrifuge finishes, carefully remove the rotor, ensuring to remove both the funnel and the filter to avoid damaging the cells.

Q & A

What is the purpose of flattening the site before observing it under a microscope?

Flattening the site allows for easier visibility of the cells under the microscope, improving the quality of the observation.

Why is it important to remove the vacuum seal before opening the rotor?

Removing the vacuum seal before opening the rotor is crucial to avoid damaging the rotor's spindle, which could disrupt the centrifuge’s function.

How can you ensure the rotor's vacuum seal is properly set?

You can check that the seal is properly set by pulling on the blue handle. If it remains in place, the seal is secure.

What is the role of the frost tent in preparing microscope slides?

The frost tent is used to position the slide correctly in the metal cassette, ensuring proper alignment of the slide for effective centrifugation.

Why is it necessary to align all parts of the cassette properly?

Proper alignment is essential to prevent any part of the holes from overlapping, which could disrupt the centrifugation process and affect the results.

What is the purpose of the filter in the slide preparation process?

The filter is placed on top of the slide to ensure that cells are evenly distributed and do not get damaged during the centrifugation.

What should be done after pipetting a 100 microliter sample into the funnel?

After pipetting the sample, place the prepared slide into the rotor with the funnel side facing the center, ensuring proper balance with another slide across from it.

Why is balancing the slides in the rotor important?

Balancing the slides ensures that the centrifuge operates evenly and prevents any potential mechanical issues during the spinning process.

What settings are ideal for spinning eukaryotic cells in the centrifuge?

For normal eukaryotic cells, the ideal settings are 800 rpm for 3 minutes.

What should be done after the centrifuge finishes spinning?

Once the centrifuge finishes spinning and a jingle sounds, open the machine, remove the rotor, and carefully extract the slide from the casing, making sure to remove the funnel and filter without damaging the cells.