Table 2.
Histochemical and immunohistochemical analyses of the intraoral pigmented low-grade AdCaNOS
| Ductal/glandular structures | Solid areas | Tumor stroma | Pigment | Details | |
|---|---|---|---|---|---|
| Histochemical analysis | |||||
| Periodic acid Schiff stain with diastase digestion | – | – | – | – | Foci of intraluminal mucinous material positive |
| Periodic-acid Schiff stain without diastase digestion | – | – | – | – | Foci of intraluminal mucinous material positive |
| Mucicarmine stain | – | – | – | – | Foci of intraluminal mucinous material positive |
| Colloidal iron staining | – | – | + focal | – | Foci of intraluminal mucinous material positive |
| Perls stain | – | – | – | – | Foci of hemosiderin granules positive |
| Fontana Masson stain | – | – | – | 1+ | Potassium permanganate bleaching-sensitive |
| Alcian blue stain | – | – | + focal | – | Foci of intraluminal mucinous material positive |
| Ziehl-Neelsen stain | – | – | – | – | |
|
Immunohistochemical analysis Antibody (Clone; dilution; manufacturer) | |||||
| pan-CK (Clones AE1/AE3; 1:500; Dako#) | 3+ | 3+ | |||
| CK7 (Clone OV-TL 12/30; 1:500; Dako#) | 3+ | 1+ | |||
| CK20 (Clone Ks20.8; 1:500; Dako#) | – | – | |||
| Vimentin (Clone Vim 3B4; 1:500; Dako#) | – | 3+ | |||
| α-SMA (Clone 1A4; 1:500; Dako#) | – | – | Scarce positive cells in close relationship with residual salivary gland ducts | ||
| Calponin (CALP; 1:300; Abcam+) | – | 1+ | |||
| h-Caldesmon (E-89; 1:40; Cell Marque♰) | – | – | |||
| DOG-1 (K9; 1:100; Leica*) | – | 1+ | |||
| 34betaE12 CK (34Be12; 1:300; Dako#) | 3+ | 2+ | |||
| S-100 (Polyclonal; 1:3000; Leica*) | 3+ | 3+ | |||
| GFAP (Clone 6F2; 1:500; Dako#) | – | 1+ | |||
| Podoplanin (Clone D2-40; 1:500; Dako#) | – | 1+ | |||
| EMA (Clone E29; 1:500; Dako#) | 2+ | – | Scarce positive cells in solid areas | ||
| p63 (Clone DAK-p63; 1:500; Dako#) | – | 3+ | |||
| CD68 (Clone KP1; 1:3000; Dako#) | – | – | |||
| CD163 (Clone 10D6; 1:500; Leica*) | – | – | |||
| CD11c (Clone 5D11; 1:500; Leica*) | – | – | |||
| CD56 (Clone CD564; 1:100; Leica*) | 1+ | 3+ | |||
| CD57 (Clone TB01; 1:600; Dako#) | – | – | |||
| ERBB2/HER2 (SP3; 1:200; Cell Marque♰) | – | – | |||
| HMB45 (Clone HMB45; 1:500; Dako#) | 1+ | 1+ | |||
| MART-1/Melan-A (A103-MART-1; 1:120; Cell Marque♰) | 1+ | 1+ | |||
| p53 (Clone DO-7; 1:500; Leica*) | – | ||||
| Ki-67 (Clone SP6; 1:500; Abcam+) | < 2% | ||||
Negative: –; Positive 1+: 5–25% of the tumor cells positive; Positive 2+: > 25–75%; Positive 3+: > 75%
Antigen retrieval was performed using 10 mmol/l citrate buffer (pH 6.0)
*Leica Biosystems Newcastle Ltd; Newcastle, United Kingdom
#Dako; Agilent Technologies, Inc., Glostrup, Denmark
+Abcam; Cambridge, United Kingdom
♰Cell Marque; Sigma Aldrich Company, Darmstadt, Germany