MicrobeHunter.com Microscopy Forum

Clear explanations, info and drawings.

http://www.olympusmicro.com/primer/anat ... rsion.html

Very clear tutorial, thanks for the link and 'heads up' to visit that Olympus primer,75RR.

I ask everyone to advise me...how do you 'oil bridge' the condenser to the specimen glass with inverted microscopes?

Or is it that only glass slides with cover slips..but 'inverted glass slides' can have an inverted microscopes condenser oiled to that specimen slide? The Olympus primer does mention oil bridge with inverted stands..but does not illustrate the setup.

I always thought ( I have a vintage Olympus CKa inverted microscope) these scopes are used with culture dishes in which case the condenser is far far away from the flat culture dish...so no way to oil bridge condenser to culture flask. So does this mean only NA of 9.0 with inverted stand objectives...and never: NA1.25-1.40?

I know inverted stands are used in thin metal/silico disc w chips etc..but I still thought the setups have the condenser far from the inverted stands specimen stage.

Again, thanks for this link,75RR. charlie guevara

I ask everyone to advise me... how do you 'oil bridge' the condenser to the specimen glass with inverted microscopes?

I have no experience with inverted microscopes - however I hope these pdfs can help

http://www.alanwood.net/downloads/olymp ... ctions.pdf
http://www.olympusamerica.com/files/seg ... oscope.pdf

Charlie g,

I have not heard of inverted objective that is more powerful than 60x NA 0.85 - most if not all are at or below 40x NA 0.65. So probably no point of using oil immersion on any side of a sample slide.

If you oil condenser, you would void intrinsic advantage of an inverted scope (which is to provide more working space above the sample/slide). And as you already know, condensers for inverted scopes are of long working distance type, so by design, it is difficult/expensive achieve high NA and LWD at the same time, even if it makes sense to do so.

thanks 75RR and zzffnn...the excellent Olympus tutorial which this thread by 75RR gives link to...it speciffically mentions imersion oiled condensers in inverted stands...hmm?!!

Like you, zzffnn, I did not think there were high NA inverted stands...yet that Olympus primer notes instances of oil bridge to condenser in inverted stands. all the best...snow on the ground and 35 degrees here (finger lakes,US) this 5:30 am...yikes...seems like a record for early snow! charlie guevara

Charlie,

I did some reading again and realized that I totally forgot live cell (fluorescence) time lapse imaging. My apologies. In that case, you do use oil immersion objectives on inverted scope with inverted slide/cover slip (0.17 mm cover slip facing down towards oiled objective) and you can go up to objective NA 1.4 on an inverted scope.

Please take a look at this PDF (especially page 1 last paragraph, figure 1 and page 3 "methods"):

https://www.google.com/url?sa=t&source= ... FrwfTZL5ww

In that case, oil immersion on objective side provides more light gathering (important for fluorescence time lapse videos) and better resolution. For observing live cells in time lapse mode, you have to keep cells naturally alive for extended time (cells have to grow at 0.17mm bottom of culture vessel with culture media), which can be done better with specialized inverted scopes like this:

http://www.vanderbilt.edu/viibre/automa ... oscopy.php

And using specialized culture vessel with #1.5 sized (0.17mm) bottom like this:
http://216.92.84.132/wordpress/product/ ... cessories/

I am not sure whether oil immersion is required on the condenser side, as I have not done such experiements myself. I would think since a layer of cell culture media is involved, oiled condenser would be cumbersome. For objective NA of 1.25, dry condenser NA of 0.95 may be enough for some cases. But specialized oiled condenser may still be used, since we are talking about super expensive specialized professional scope system - anything is possible and no expense is saved here.

Wow, zzffnn!!! I went to thathose links, what intricate devices for live cell microscopy! Heaters which attach to the objective barrel to match approx 37 C temp of live cells...and the advice to use Cargille immersion oil of a specific blend to at 37 C have RI of 1.15...! And those culture dishes..I wonder how much training needed just to have a sense of what equipment is available for a given live cell microscopy project...I wonder which training institutions keep up to date with all that instrumentation...and the need to be: 'good with your hands' to use those setups!

Thanks for the link to: Biotech,PA,US charlie guevara

Charlie g,

I am impressed by the fact that you read everything. I read those pages very quickly and missed most things that you mentioned.

RI 1.15 oil blend? That would require expensive water immersion objectives, I guess (unless their infinity scope is dramactically modified to take LOMO WIs LOL). I suspect they don't use water immersion (but use oil blend to achieve the same RI) to avoid evaporation, vibration and mold?

Good to know. But too expensive for us hobbyists.