5 replies to this topic

[Helios]

Has anyone used laab-tek chambered coverglass (cat no. 155380) for confocal microscopy???
after dapi staining, do you need to cover the cells with coverslip or can you directly visualise them under the confocal microscope?

[bob1]

If you want to use oil immersion lenses (20x or higher objective) you need to coverslip the slide so you can put the oil on it.  With lower mags, it may not be absolutely necessary, but you don't want the lens touching your sample or the lens getting covered in stuff from your sample.  Basically - always coverslip your slides.

[SuMi]

You generally have to use coverslips for confocal as the thickness of the coverslip is taken into consideration. We were told to always use No. 1.5 coverslips as this gives the most accurate result.

[Prep!]

yeah i vaguly remember having heard that the cover slip thickness also plays a rle in magnification!!! though wat bob says also is very logical!!!

P.S. its been ages since i even saw a microscope!!!     

[rhombus]

Pradeep Iyer, on Dec 2 2009, 09:50 AM, said:

yeah i vaguly remember having heard that the cover slip thickness also plays a rle in magnification!!! though wat bob says also is very logical!!!

P.S. its been ages since i even saw a microscope!!!     

Dear Pradeep Iyer,

You are probably referring to the "working distance " of the objective lense rather than magnification itself. Most confocal microscopes are purchased with long working distance objectives so that the thickness of nay coverslip used will not be an issue.

Kindest regards

Rhombus

[Prep!]

rhombus, on Dec 2 2009, 06:03 PM, said:

Pradeep Iyer, on Dec 2 2009, 09:50 AM, said:

yeah i vaguly remember having heard that the cover slip thickness also plays a rle in magnification!!! though wat bob says also is very logical!!!

P.S. its been ages since i even saw a microscope!!!     

Dear Pradeep Iyer,

You are probably referring to the "working distance " of the objective lense rather than magnification itself. Most confocal microscopes are purchased with long working distance objectives so that the thickness of nay coverslip used will not be an issue.

Kindest regards

Rhombus

Thank you for teh enlightenment rhombus!!!