Is it suitable to use scanning electron microscope (SEM), or is there any antibody to show gold nano inside the cell?
the best method to observe nanoparticles within the cells is transmission electron microscopy, SEM only scans the surface giving the surface topography, however electron beam penetrates in the cells transmit from cells. giving more detailed information about the sample , hence the nano particles within the cell.
Gaser is right, both these methods will work, however, the rational or endpoint for your imaging will dictate which method is best for you. If you want an overall estimation of the total amount of nanoparticles within your cells, the laser excitation method will be the most useful, but the limitation of this approach is the resolution (you will not be able to see organelle specific localisation. If it is high detail you require TEM (although time consuming) is the best option. There is also the option of hyper-spectral imaging in Live cells using the Cytoviva microscope if you have access to this instrument.
Hope this helps
In our Lab we use CytoViva hyperspectral. Excellent for viewing gold nanoparticles in cells. However you need to form a hypersctral library of the gold nanoparticles.
TEM or HR-TEM is the best option madam.... so that you can study them clearly..
you can do intracellular quantification (down to 35 femtogram, equivalent to single nanoparticle) perfectly with standard confocal laser scanning microscope. For 60 nm, the results perfectly correlate to mass specrtometry. For particles around 40 nm (the size you want to use), you still can quantify, but you will unterestimate the absolute concentration. This os not a big deal if you have calibrated before. Details in the CLSM filter and wavelegth settings you can find here:
Klein, S. ; Petersen, S. ; Taylor, U. ; Rath, D. ; BARCIKOWSKI, S.: Quantitative visualisation of colloidal and intracellular gold nanoparticles by confocal microscopy. In: Journal of Biomedical Optics 15 (2010), Nr. 3, S. 036015
Collect the cells, process them into Spur resin, ultrasection them and see the particles with electron micrscopy
SERS is very suitable for this purpose, the publication in attachment:
Article Magnetic Gold Nanoparticles in SERS-Based Sandwich Immunoass...
As Iva Que said fluorophor labelling is a simple way to do it. Especially, with FITC labelling you are able to see the cell uptake of GNPs using a fluoroscence microscopy (with green filter). FITC can bind GNPs directly, there are some reported works about it. Good luck!
the best method to observe nanoparticles within the cells is transmission electron microscopy, SEM only scans the surface giving the surface topography, however electron beam penetrates in the cells transmit from cells. giving more detailed information about the sample , hence the nano particles within the cell.
How to add GNPs uptaken cells in TEM grid? Can anybody suggest me some papers for TEM imaging of GNPs cell uptake protocol.?
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