Leica Novocastra Liquid Mode D'emploi page 4

Quality Control
Differences in tissue processing and technical procedures in the user's laboratory may produce significant variability in results,
necessitating regular performance of in-house controls in addition to the following procedures.
Controls should be fresh autopsy/biopsy/surgical specimens, formalin-fixed, processed and paraffin wax-embedded as soon as possible
in the same manner as the patient sample(s).
Positive Tissue Control
Used to indicate correctly prepared tissues and proper staining techniques.
One positive tissue control should be included for each set of test conditions in each staining run.
A tissue with weak positive staining is more suitable than a tissue with strong positive staining for optimal quality control and to detect
minor levels of reagent degradation.
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Recommended positive control tissue is colon.
If the positive tissue control fails to demonstrate positive staining, results with the test specimens should be considered invalid.
Negative Tissue Control
Should be examined after the positive tissue control to verify the specificity of the labeling of the target antigen by the primary antibody.
Recommended negative control tissue is cerebellum.
Alternatively, the variety of different cell types present in most tissue sections frequently offers negative control sites, but this should be
verified by the user.
Non-specific staining, if present, usually has a diffuse appearance. Sporadic staining of connective tissue may also be observed in
sections from excessively formalin-fixed tissues. Use intact cells for interpretation of staining results. Necrotic or degenerated cells often
stain non-specifically. False-positive results may be seen due to non-immunological binding of proteins or substrate reaction products.
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They may also be caused by endogenous enzymes such as pseudoperoxidase (erythrocytes), endogenous peroxidase
(cytochrome C), or endogenous biotin (eg. liver, breast, brain, kidney) depending on the type of immunostain used. To differentiate
endogenous enzyme activity or non-specific binding of enzymes from specific immunoreactivity, additional patient tissues may be stained
exclusively with substrate chromogen or enzyme complexes (avidin-biotin, streptavidin, labeled polymer) and substrate-chromogen,
respectively. If specific staining occurs in the negative tissue control, results with the patient specimens should be considered invalid.
Negative Reagent Control
Use a non-specific negative reagent control in place of the primary antibody with a section of each patient specimen to evaluate
non-specific staining and allow better interpretation of specific staining at the antigen site.
Patient Tissue
Examine patient specimens stained with NCL-L-CEA-609 last. Positive staining intensity should be assessed within the context of any
non-specific background staining of the negative reagent control. As with any immunohistochemical test, a negative result means that
the antigen was not detected, not that the antigen was absent in the cells/tissue assayed. If necessary, use a panel of antibodies to
identify false-negative reactions.
Results Expected
Normal Tissues
Clone COL-1 detected the carcinoembryonic antigen in epithelial cells of the colon, rectum, esophagus and larynx, and in the squamous
mucosa of the cervix. (Total number of normal cases = 150).
Abnormal Tissues
Clone COL-1 stained 58/65 colon tumors (including 20/23 metastatic adenocarcinomas, 18/22 adenocarcinomas, 6/6 adenomas,
5/5 polyps, 4/4 mucinous adenocarcinomas, 3/3 metastatic mucinous adenocarcinomas and 2/2 metastatic signet-ring cell carcinomas),
33/77 lung tumors (including 14/28 adenocarcinomas, 12/28 squamous cell carcinomas, 3/8 small cell carcinomas, 1/6 atypical
carcinoids, 1/3 large cell carcinomas, 1/2 metastatic squamous cell carcinomas and 1/2 metastatic adenocarcinomas), 10/49 ovarian
tumors (including 3/12 endometrioid adenocarcinomas, 3/3 mucinous adenocarcinomas, 2/6 serous papillary adenocarcinomas,
1/3 mucinous cystadenocarcinomas, 1/3 mucinous cystadenomas, 0/5 serous adenocarcinomas, 0/4 theca cell tumors, 0/3 granulosa
cell tumors, 0/3 undifferentiated carcinomas, 0/2 endodermal sinus carcinomas, 0/1 dysgerminoma, 0/1 immature teratoma,
0/1 leiomyosarcoma, 0/1 borderline mucinous cystadenoma and 0/1 adenocarcinoma), 5/11 metastatic tumors (including 3/4
metastatic gastric carcinomas, 2/4 metastatic breast carcinomas, 0/1 metastatic nasopharyngeal carcinoma, 0/1 metastatic thyroid
adenocarcinoma and 0/1 metastatic esophageal squamous cell carcinoma), 5/5 chronic colitis, 4/4 Crohn's disease, 3/3 stomach
adenocarcinomas, 2/2 tumors of the small intestine (including 1/1 adenoma and 1/1 adenocarcinoma), 2/2 squamous cell carcinomas of
the cervix, 1/3 esophageal squamous cell carcinomas, 1/2 endometrial adenocarcinomas, 1/1 squamous cell carcinoma of the skin and
1/1 squamous cell carcinoma of the tongue. No staining was observed in breast tumors (0/5), thyroid tumors (0/5), brain tumors (0/4),
hepatocellular carcinomas (0/4), lymphomas (0/3), prostatic tumors (0/2), tumors of the head and neck (0/2), tumors of the adrenal gland
(0/2), bladder tumors (0/2), bone tumors (0/2), tumors of the salivary gland (0/2), seminomas (0/2), kidney tumors (0/2), a pancreatic
tumor (0/1), a prostatic hyperplasia (0/1), a melanoma (0/1), a case of tuberculosis of the colon (0/1) and a pheochromocytoma (0/1).
(Total number of abnormal cases = 267).
NCL-L-CEA-609 is recommended for the detection of carcinoembryonic antigen in normal and neoplastic tissues, as an adjunct
to conventional histopathology using non-immunologic histochemical stains.
CEA-609-L-CE
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