Watch our video for the counterstain and probe analysis step of the CytoCell® hematology FISH protocol. Study the Package Insert (Instructions for Use, IFU) carefully before using the protocol depicted in this video. This video does not replace the content from the Package Insert. The Package Insert can be found in the product packaging of each FISH Probe Kit and in the Product Documentation section of each FISH Probe Kit on the website. Refer to the Package Insert for warnings, precautions, storage and handling.
Watch quick video tips and tricks from our OGT experts for the counterstain and analysis stage of the CytoCell hematology FISH protocol.
Try to use just one type of immersion oil in your laboratory; different types of oil may be immiscible. Immersion oil of one type remaining on a microscope objective may result in a ‘milky’ appearance and a reduction of light transmission when it comes in contact with a slide with a different type of immersion oil.
Once the DAPI counterstain and coverslip have been applied to the slide, it is recommended that users seal the coverslip with varnish. This prevents excessive drying out and maintains the probe signal strength. For short term storage (0-2 weeks), the slides should be covered and stored at room temperature. For long term storage, the slides should be stored 2-8°C and covered.
Currently, there are two different triple filters commonly used in laboratories, one which is specific for TRITC/FITC/DAPI and one which allows visualization of Texas Red/FITC/DAPI. The first one is easily distinguished by the green light that comes from the objective on the microscope, the other gives a white/pale yellow colour. The only filter that allows simultaneous visualization of CytoCell's green, red and blue dyes is a Texas Red/FITC/DAPI filter.
Both types of filter can be used, but using the Spectrum Red will give brighter signals for Texas Red probes and using the Spectrum Orange will give brighter signals for our orange fluorophore.
Depending on the manufacturer’s recommendations, fluorescent filters typically need to be replaced every 2-4 years. Over time and use, the filter will begin to degrade. The damaged emission/excitation filters will show a mottled appearance, which begins on the edges and will eventually completely obscure the filter. The effects may not be noticeable on a day to day basis, but will manifest as higher than normal background and weaker signals.
Please contact CytoCell technical support for further advice and guidance if you suspect filter problems.
Depending on the type of bulb used, the shelf life can vary from 200-3000 hours. Please verify with your supplier. The older the bulb, the dimmer the fluorescence signals become, and the greater the chance of the bulb exploding. This is dangerous as mercury vapor is highly toxic.