With the help of the talented machinists at Caltech, I made this quintuple dichromatic beamsplitter cube slider. It allows the user to easily change between five excitation / emission wavelength combinations.
An aluminum extension (not yet anodized) must be made and inserted to allow the dichroic beamsplitter / filter slider to pass under the angled objective turret.
Note: the adjustable stage must be raised but the focus adjustment easily compensates for the slight extension. Relatively inexpensive "stage micrometers" (microscope slide with a metallic rule subdivided into ten micrometer divisions) are available from Scientific Vendors and are indispensable aids for photomicroscopy.
Often fluorescence microscopy is conducted in a darkened room to allow the microscopist's eyes to "dark adapt" so that faint fluorescence can be seen. In "dark adaptation" the pupils dilate and the retina's pigment epithelium retracts. This is especially useful when "picking clones" that are expressing green fluorescent protein (GFP).
There can be significant amounts of ultraviolet light pollution where mercury illumination is being used. Unseen ultraviolet was recorded as purple in this "dark-field" photograph of the microscope to the left. Ultraviolet light can not only contribute to cataracts, sunburn, snowblindness, skin cancer, it can also degrade images; especially during long exposures.
Make sure that sources of ultraviolet light such as germicidal mercury vapor lamps are turned off. Wear protective goggles or glasses, if possible.
The orange eye guard can be attached using adhesive Velcro strips to prevent its being accidentally dislodged and broken.
List of useful microscopy sites
- Zeiss Microscopy & Image Analysis:
- Confocal Introduction Washington/Keck
- Microscopy Primer Objectives
- An "E-zine" resource: Microscopy Online
- Commercial Dichroic Mirrors ("Filters") / GFP Chroma
- Commercial Fluorescent Labels Clontech
- UltrafastSensors ... Ultrafast Sensors
- The Microscope Lab ... Extensive - with many Links
