Microscope, Nikon Optiphot Planar Microscope

Attachments

Prepared by:

Ying Qi & Justin Scanlon

Location:

Dow 2224

Hazards:

The following materials and equipment associated with this procedure present exposure or physical health hazards. Safety precautions are prudent and mandatory.

None

Controls:

Prior to performing this procedure, the following safety equipment must be accessible and ready for use: (e.g. chemical fume hood, biological safety cabinet, laminar flow hood, chemical spill kits)

None

Protective Equipment:

Prior to performing this procedure, the following personal protective equipment must be obtained and ready for use: (e.g. acid resistant gloves, safety eyewear, lab coat, chemical splash apron)

None

Waste Disposal:

This procedure will result in the follow regulated waste which must be disposed of in compliance with environmental regulations.

None

Spill:

In the event that a hazardous material spills during this procedure, be prepared to execute the emergency procedure below.

Not Applicable

Instructions:

OLD SOP

General instructions:

Remove the dust cover.
Turn on the power source for either transmitted or reflective light. Do not use an excessively low voltage, use at least 6V or higher.
To switch from looking in the eyepieces to capturing images, rotate the eyepieces clockwise.
For instructions on capturing images for the various techniques, please see their SOP by clicking the link:
- For using the Nikon Digital Insight DS-L1 (SOP): (the corresponding online systems if the units are turned onSorby.engin.umich.edu, Sauveur.engin.umich.edu)
- For using the Nikon Digital Insight DS-L2 (SOP): (the corresponding online system if the unit is turned onNomarski.engin.umich.edu)
- For using the Image J software on the Apple G4 computer for the Cohu high performance CCD camera (SOP)
Using the transmitted light source for capturing images is similar to the steps below (except where noted)

For focusing the sample:

Start with a large sample distance and move closer. Always start on the objective lens with the lowest magnification (5X on the microscopes in VVUL). These steps will prevent accidentally crashing into your sample damage to the lens or your specimen by crashing into it. There are 4 different objective lenses for the Optiphots in the VVUL: 5X, 10X, 40X and 100X (kept in Dow 2224C)
There is a 10X multiplier both in the eyepieces and in the image capturing devices,(meaning the total magnification for the 5X is actually 50X, and so on) To prevent damage to the lenses, use the revolving nosepiece to rotate between lenses.
Adjust the focal point by using the course adjustment knob and then with the fine adjustment knob.

Field and Aperture Diaphragm:

Field:

The field diaphragm is used to restrict the illumination range (observation range). The residual diffused reflected light occurring on the specimen or lens is restricted by stopping the field diaphragm down to the required range. As a result, flare is reduced and a high contrast specimen image can be observed.*

Aperture:

The aperture diaphragm is attached to the condenser lens and used to control resolving power, contrast and depth of focus. However, these factors cannot be controlled independently.*

Resolving Power

Contrast

Depth of focus

Brightness

Aperture diaphragm open 100%

High

Low

Shallow

Bright

Aperture diaphragm stopped down

Low

High

Deep

Dark

Rings for Reflective light microscope

Transmitted Light Field and Apeture Rings

Rings for transmitive light microscope

Use of polarized microscopy:

These are the transmitted light polarizer set, it is used in conjunction with the analyzer for simplified polarizing observation.
Polarized light allows for the observation of:
- Interference patterns
- Stress fields
- Birefringence
- Refractive index differences
- Optic relief of the specimen
For more information go to MicroscopyU's page about polarized light:
http://www.microscopyu.com/articles/polarized/polarizedintro.html
Reflective light polarizer sets are found in Dow 2224C

Use of filters:
ND32 is used for brightness adjustments

GIF filter is used for contrast adjustment, mainly in B/W imaging
For darkfield microscopy:
- The mirror-change knob should be pulled out.
- Both diaphragms should be wide open.
- Place in any filters to be used in slots provided in front of the transmitted light source.
- Make certain there are no polarizing plates or a Nomarski prism in the light path.
- Adjust the brightness by changing the lamp voltage.
- Continue to follow the procedure for the appropriate image capturing device
For brightfield microscopy:
- The mirror-change knob should be pushed in.
- Place in any filters to be used in slots provided in front of the transmitted light source.
- If using the Nomarski prism or any polarizing plates place them in their respective locations.
- Adjust the brightness by changing the lamp voltage.
- Continue to follow the procedure for the appropriate image capturing device.