Recently, I am preparing multi-compartment liposomes（MIVs）. I want to observe it with a confocal microscope or a phase contrast fluorescence microscope, but I don’t know how to prepare the sample.
I tried to mix 1:1 with glycerol and buffer before, and then drip the liposome sample on the glass slide, then drop the above liquid on the glass slide and mix with the sample to be tested. However, after putting on a cover glass and observing with a microscope after mounting, the liposomes had been broken.
My liposomes are positively charged liposomes.
I am just venturing into this field of research and any advice on this topic would be highly appreciated.
Hi,
Attached literature describe different microscopic techniques for liposome analysis and how to prepare samples for observations.
More info:
Try the following procedure:
2.7. Confocal scanning laser microscopy (CSLM):
Samples of nanoliposomes were mixed with Nile Blue dye
(BDH, England, minimum dye content 90%, 1 g/l). A small
aliquot of the sample (usually 10–20 ml) was then transferred to
a glass slide which was then covered with a coverslip. CSLM
was performed on a Leica TCS 4D confocal microscope (Leica
Lasertechnik GmbH, Heidelberg, Germany) with 40and
100 oil immersion objectives (numerical apertures 1.3 and
1.4, respectively). The microscope was used in a fluorescent
mode at room temperature. Images were obtained using an aircooled
Ar/Kr laser at 488 nm argon laser (5 mW) at the
excitation and emission wavelengths of 543 and 580 nm.
From:
Colas, J. C., Shi, W., Rao, V. M., Omri, A., Mozafari, M. R., & Singh, H. (2007). Microscopical investigations of nisin-loaded nanoliposomes prepared by Mozafari method and their bacterial targeting. Micron, 38(8), 841-847.
I will be following this with great interest. I am not a microscopy expert, but I would be asking myself what broke them, the mixing, dissolved, and or the physical stress/pressure of the glass on top. I just downloaded all the files so I guess I have some reading to do. The MCL topic is new to me, and has tweaked my interest.
Dear Mike McGinness
The world of microscopy is fascinating and wonderful indeed. As we advance in science and technology, we have better tools to visualize, directly or indirectly, smaller and smaller objects.
Perhaps the following papers will interest you. Have a look and let me know your thoughts or if you like to have a PDF:
The role of high-resolution imaging in the evaluation of nanosystems for bioactive encapsulation and targeted nanotherapy
K Khosravi-Darani, A Pardakhty, H Honarpisheh, V Rao, MR Mozafari
Micron 38 (8), 804-818
------------ ---------------
Microscopy in nanobiotechnology
MR Mozafari
Micron 38 (8), 775-776
-----------------------------
Ultrastructural architecture of liposome-entrapped glutathione: A cryo-SEM study
MR Mozafari, CJ Reed, C Rostron
Cell. Mol. Biol. Lett 9, 101-103
----- ----- -----
A review of scanning probe microscopy investigations of liposome-DNA complexes
MR Mozafari, CJ Reed, C Rostron, V Hasirci
Journal of liposome research 15 (1-2), 93-107
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