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Having Probelems?

Complication:

DIFFUSE PROBE SIGNALS
This may be related to high-homology regions or inconsistent cell lines.


Solution:

If diffuse signals are on a cell line, run probe on normal sample to rule out a cell line mutation.

Complication:

NO SPECIFIC SIGNALS & HAZY
Underdigested tissue samples can prevent probe from entering nuclei.


Solution:

Increase digestion time or protease concentration to eliminate extracellular tissue. Cells should appear dark gray and distinguishable under a light microscope.

Complication:

HIGH LEVELS OF BACKGROUND
Non-stringent washes can cause background fluorescence in cells and make signal detection difficult.


Solution:

Add additional wash steps, or repeat post hybridization washes. If issue persists, decrease hybridization time.

Complication:

SMALL, WEAK SIGNALS
Small and faint signals can result from improper sample preparation, incorrect filter specifications, or non-optimal hybridization conditions.


Solution:

Ensure fixative was prepared properly and filter specifications are compatible with Empire Genomics’ probes. Time and temperature of hybridization may need to be adjusted as well.

Complication:

CELLULAR DEBRIS & BACKGROUND
If samples have not been properly fixed, inner nuclei staining or cellular debris can remain in the sample.


Solution:

Make sure to use fresh samples and fixative. Both reagents and glassware should be changed regularly.