As I wrote earlier that I am bascially an engineer and
trying to use confocal microscopy to study the
diffusion across microbial floc obtained in a
wastewater treatment plant. This particular microscope
is being used for the first time in my lab., I am
finding it difficult to figure out. I have gone
through all the suggestions I received in this forum.
Although, it may look very basic and trivial and
perhaps not worthy to put in for discussins, I hope,
to receive answers to my queries:
I have LSM 510 Zeiss Axioplan 2 machine. I click on
expert mode and click on 'Micro' and get on the screen
which says, "Axioplan". Now, it has got different
headings:
1. 'Refelected light', here, I have option of: 'on'
but aperture n.a.. So, I have just two options, either
to put reflected light on or off. Could you tell me
when it has to put on and when not? Again it works
only when the microscope is in VIS mode and not when
in LSM mode.
2. Next comes, 'Reflector turret'. Here, it has got
DIC, Fset 09wf, Fset15 wf, None, None. Now, which is
to be used and in what combination. And, why there is
two 'None'?
3. 'Objective': There are five objectives and one
empty position. How to decide which one to be used?
4. 'Condensor': It is fixed at aperture 0.05 with any
changes n.a., so this saves me from deciding about any
figure.
5. 'Filter': This adjusted in terms of % transmission.
Now, how to decide about what percentage of llight is
to be used?
6. 'Field Stop': To be selected in terms of % field
stop. How to fix up the figure?
7. 'Transmitted light': to be selected in intersity %.
Again how to fix up the figure?
Further, when I start with zero % transmission filter,
zero % intesity of transmitted light and zero % field
shop and any reflector turret at the VIS (or with any
other value) but when I pull out the rod for LSM,
filter comes to 12 %, transmitted light to 4 % , field
stop at 12.5% and reflector turret to none position.
Should I assume that LSM mode selects these values on
its own and we should not try to set these values.
These questions may appear to be quite childish to the
experts but I think, they are very useful to the
biginners and will be useful in future, too.
--Bhuwan
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