| Problem | Cause | Solution |
|---|---|---|
| Weak or no Staining | 1. Low concentration of specific oligosaccharide on sample. 2. Low concentration of lectin conjugate. 3. Low concentration of Dye/Substrate. 4. Insufficient incubation time. 5. Inappropriate treatment of sample prior to labeling. | Causes #1 - #4 a. Increase incubation time. b. Increase concentration of sample (on blot), lectin conjugate and/or Dye/Substrate. a. Treat section or blot with a different blocking reagent. |
| High Background | 1. Lectin conjugate and/or Dye/Substrate is too concentrated. 2. Insufficient washing. 3. Insufficient blocking. 4. Sample contains enzymatic activity. | a. Decrease concentration of respective reagents. b. Shorten incubation time. a. Perform multiple washings and prolong washing time. a. Treat section or blot with a different blocking reagent. a. Determine if the sample contains activity which would give background staining in the absence of the Dye/Substrate. |
| Unexpected Staining Pattern | Multiple causes | a. Perform control reactions. b. Use other cytochemical techniques to prove or disprove finding. |
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