I am currently trying to isolate Haematococcus pluvialis microzoid (~10micron) using a Pasteur pipette pulled into a micropipette and a light microscope. However I have a hard time to detect my micropipette under the microscope. Can anyone suggest how to get a gentle suction to get the desired cell, too?
Hi Tan Wai Yee;
I would suggest either: 1) attach some tubing to the pipet and use your own lungs to suck the cells into the pipette. If you do this, it is best to draw a little bit of deionized water into the pipette tip prior to isolation. OR 2) use your thumb over the back of the pipet to prevent any water coming in and once your pipette tip is aligned with the specimen you can release your thumb and the small amount of vacuum can draw the cells in.
Method 1 has been very effective for me. If you need some clarification I could try to provide some photos of the setup.
Dear David, thank you for your answer. I would like to know how do you settle the problem of not able to see the tip under the microscope? I tried to adjust the knobs but I only managed to get a shadow-like image.
It would be so much help if you can provide some photos of your setup! Many thanks!
Hello Tan Wai Yee,
I did a lot of microalgae isolations before using the same method David R. L. Burge refers (method 1). I would turn a glass Pasteur pipette even thinner using a flame to turn the tip into a really small capillary and attach a rubber tube at the end to use my lungs to help with the process. I would use an Inverted Optic Microscope which helps even more with the handling during the isolations and because with these microscopes the light comes from above the sample. So, as Chee Ping Tan, when I had located the microalgae cell I wanted to isolate, I would start to bring my capillary from above the sample until I could locate its shadow without touching the sample yet. When located, it was just a matter of making contact on the sample and you would see the tip of your capillary sucking the cell you are trying to isolate. Then yes, I would use my lungs to transfer the cell from the pipette into a multi-well plate with liquid medium.
At the beginning it can be quite frustrating because it’s exhausting and you’re not as accurate as you wanted it, but after a few sessions you will get much better and even develop your own tricks.
Good luck
Dear david ji,
You can use inverted microscope or stereo zoom microscope for isolation purpose where you will get enough work space..it will make ur job easier..
With regards...
Perumal
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