The lung tissue sections were de-paraffinized and rehydrated according to standard protocols. After antigen retrieval in 0.01 M citrate buffer (pH 6.0), the tissue sections were incubated with 1% Triton X-100 for 15 min and incubated with 5% BSA (in TBS, pH=7.4) for 30 min, followed by overnight incubation with the primary antibodies at 4 °C. After a wash with TBS, the sections were incubated with the secondary antibodies at room temperature for 1 h. After being washed three times with TBS, the sections were mounted using the 496-diamidino-2-phenylindole (DAPI) mounting medium. The fluorescence was recorded with a ZEISS Z1 AxioObserver fluorescence microscope (Carl Zeiss, Oberkochen, Germany).
Z1 axioobserver fluorescence microscope
Manufactured by Zeiss
Sourced in Germany
The ZEISS Z1 AxioObserver is a fluorescence microscope designed for live-cell imaging and high-resolution analysis. It features a fully automated platform with motorized components, enabling precise control of sample positioning and imaging parameters. The microscope is equipped with a selection of fluorescence filter cubes, providing the capability to visualize various fluorescently labeled samples.
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Lab products found in correlation
2 protocols using z1 axioobserver fluorescence microscope
1
Immunofluorescence Staining of Lung Tissue
2
Immunofluorescence Analysis of Lung Tissue
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Paraffin-embedded lung tissue sections were deparaffinized with xylene and rehydrated. Antigen retrieval was employed before blocking in PBS with 10% normal goat serum, 0.1% BSA, 0.3% TX-100. The antigen retrieval solution is Tris-EDTA buffer (10 mM Tris Base, 1 mM EDTA solution, 0.05% Tween 20, pH 9.0). The dilutions for rabbit against NAMPT (Bethyl Laboratories, Inc. catalog# A300-372A) and mouse against von Willebrand factor (VWF, catalog# MA5-14029, Invitrogen) are 1:100. Secondary antibodies for NAMPT and VWF were Alexa Fluor® 593 Donkey anti-Rabbit IgG antibody (1:500) and Alexa Fluor® 593 goat anti-mouse IgG antibody (1:500), respectively. An anti-fade mounting media with DAPI (Life Science Inc) was used to fix the coverslip to a slide. The slides were examined using a a ZEISS Z1 AxioObserver fluorescence microscope (Carl Zeiss, Oberkochen, Germany).
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