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conjugated antibodies reveal mature zymogene granules in the apical region of
acinar cells in human pancreatic tissue (FITC filter setup). (B) Endocrine cells of the
islets of Langerhans in rat pancreatic tissue sections were visualized using antibodies
specific for synaptobrevin2 and Alexa Fluor® 555-conjugated secondary antibodies
(Cy3 filter setup). (C, D) Infiltrating granulocytes in human chronic inflammatory
pancreas were marked by anti-Calgranulin B antibodies and Alexa Fluor® 594-
conjugated secondary antibodies (Texas Red filter setup). A group of infiltrated
granulocytes at higher magnification is displayed in D. (bar: A-C =50 µm, D =10 µm)
Fig. 6
Toluidine Blue treatment to quench autofluorescence of human pancreatic tissue
sections compared to Sudan Black treatment. (A-C) Distribution of autofluorescence
in pancreatic section without treatment, visualized with different filter setups, as
indicated. (D-F) Comparable sections were either stained with Toluidine Blue, or (G-I)
a combined treatment with Toluidine and Sudan Black was performed, or (J-L) the
protocol for Sudan Black treatment was applied. As control for the performance of
specific immunolabeling, zymogene granules marked with anti-Reg3a antibodies and
Alexa Flour® 488-conjugated secondary antibodies were analyzed with FITC filter
setup (B, E, H, K). Note that Toluidine Blue treated sections show enhanced
fluorescence of pancreatic acini with DAPI and Cy3 filter setups (D, F) although
somewhat reduce autofluorescence was observed when using the FITC filter setup
(E). (exposure time: DAPI 50 ms; FITC 500 ms; Cy3 300 ms; objective: 40x; bar=50
µm)
Fig. 7
Quenching of autofluorescence in human pancreatic tissue sections using cupric
sulphate treatment, compared to Sudan Black application. A specific fluorescence
labeling of acinus granules was performed as described in Fig. 4. Images from
microscopic observations using FITC (A, C, E, G, I) or Cy3 (B, D, F, H, J)
epifluorescence filter systems are shown. Images obtained from control specimen
without treatment (A, B) are compared to tissue sections after incubation in 50 mM
(C, D) or 100 mM (E, F) cupric sulphate. Sections after application of the Sudan
Black protocol alone (I, J) exhibit superior results compared to combined treatments
with cupric sulphate following Sudan Black (G, H). Please note that the fluorescence
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