40 Questions
What is EasySep Direct used for?
EasySep Direct is used for isolating cells directly from whole blood without the need for RBC lysis, density gradient centrifugation, or other pre-processing steps.
What must be added to the whole blood when isolating myeloid cells?
EDTA must be added when isolating myeloid cells.
How long should the RapidSpheres be vortexed for?
The RapidSpheres should be vortexed for 30 seconds.
How should the isolation cocktail be added to the sample?
The isolation cocktail should be added using a pipette.
What should be done after adding the RapidSpheres to the sample?
The sample should be mixed using a pipette due to the viscous nature of blood.
How long should the sample be incubated at room temperature?
The sample should be incubated at room temperature according to the product information sheet, typically for 5 minutes.
What medium should be used to top up the sample for lymphoid isolations?
For lymphoid isolations, calcium and magnesium free PBS should be used to top up the sample.
How should the sample be mixed after topping up with the recommended medium?
The sample should be mixed using a pipette by drawing the sample up and down 2-3 times.
Where should the tube be placed after mixing the sample?
The tube should be placed into the magnet without a lid, ensuring it is fully seated in the indentation.
What is the final step after placing the tube into the magnet?
The final step is not provided in the text.
What is the purpose of incubating the sample at room temperature on the magnet?
To keep the magnetically labelled unwanted cells & RBCs to the back & bottom of the tube.
What should be done with the clear supernatant containing enriched cell suspension?
It must be pipetted off, not poured.
What volume of pipette should be used for the 5mL side of the magnet?
2mL pipette should be used.
How should the EasyEights magnet be placed for easier visualization?
It can be placed at a 45 degree angle.
What should be done when pipetting the supernatant off the bottom RBC pellet?
Collect the upper fraction of clear cell suspension, taking care not to disturb particles on the magnet side of the tube.
What should be done if isolating myeloid cells?
Use a fixed collection volume and refer to the info sheet for volume of cell suspension to be collected.
What should be done if isolating lymphoid cells?
Collect entire upper phase from top to bottom moving down the front of the tube, and also collect a small volume of RBC pellet (up to 10% of original sample volume).
What should be added to the new tube containing the enriched cell suspension?
RapidSpheres should be added.
What should be done after adding RapidSpheres to the new tube?
Mix with a pipette.
What is the final step to prepare enriched cells for use?
The enriched cell suspension should be transferred to a new tube.
What is the purpose of incubating the sample at room temperature on the magnet?
To keep the magnetically labeled unwanted cells and RBCs to the back and bottom of the tube, allowing the clear supernatant containing enriched cell suspension to be pipetted off.
What medium should be used to top up the sample for myeloid isolations?
A fixed collection volume with some RBCs to ensure optimal recovery, as indicated in the information sheet.
What should be added to the new tube containing the enriched cell suspension?
RapidSpheres should be added to the new tube containing the enriched cell suspension.
What should be done if isolating lymphoid cells?
Collect the entire upper phase from top to bottom moving down the front of the tube, and also collect a small volume of RBC pellet (up to 10% of the original sample volume).
What should be done after adding RapidSpheres to the new tube?
The mixture should be mixed with a pipette.
What should be done when pipetting the supernatant off the bottom RBC pellet?
Pipette the supernatant off the bottom RBC pellet, collecting the upper fraction of clear cell suspension from top to bottom, taking care not to disturb particles on the magnet side of the tube.
What is the final step to prepare enriched cells for use?
The enriched cell suspension should be transferred to a new tube and is now ready for use.
Where should the tube be placed after mixing the sample?
The tube should have the lid removed and be placed into the EasySep magnet for a second round of separation.
How long should the RapidSpheres be vortexed for?
The text does not specify a particular duration for vortexing the RapidSpheres.
How should the sample be mixed after topping up with the recommended medium?
The sample should be mixed with a pipette.
What is EasySep Direct used for?
EasySep Direct is used for isolating cells directly from whole blood without the need for RBC lysis, density gradient centrifugation, or other pre-processing steps.
What needs to be added to the round bottom tube if isolating myeloid cells?
If isolating myeloid cells, EDTA must be added to the round bottom tube.
How long should the RapidSpheres be vortexed for?
The RapidSpheres should be vortexed for 30 seconds.
What should be done after adding the isolation cocktail to the sample?
After adding the isolation cocktail to the sample, the RapidSpheres should be added to the sample using a pipette.
Why is using a pipette important when mixing the sample?
Using a pipette is important when mixing the sample because blood is viscous, and using a pipette ensures it is well mixed.
How long should the sample be incubated at room temperature?
The sample should be incubated at room temperature, with timing according to the product information sheet, typically around 5 minutes.
What should be used to top up the sample, and what are the specific recommendations for lymphoid and myeloid isolations?
The sample should be topped up with the recommended medium. For lymphoid isolations, calcium and magnesium free PBS should be used, while for myeloid isolations, calcium and magnesium free PBS with 1 mM EDTA should be used.
How should the sample be mixed after topping it up with the recommended medium?
After topping up the sample with the recommended medium, it should be mixed using a pipette by drawing the sample up and down 2-3 times.
What should be done before placing the tube into the magnet?
Before placing the tube into the magnet, the lid should be removed, and the tube should be seated fully in the indentation.
What is the purpose of the EasySep Direct platform for isolating cells?
The purpose of the EasySep Direct platform is to provide a fast, column-free, fully immunomagnetic platform for isolating cells directly from whole blood without the need for RBC lysis, density gradient centrifugation, or other pre-processing steps.
Test your knowledge of the EasySep Direct immunomagnetic platform for isolating cells directly from whole blood without the need for RBC lysis or density gradient centrifugation. Learn about the simple steps involved in isolating cells using this fast and column-free method.
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