Complete worms of diverse T. spiralis developmental stages (ML, 6h and 12h IIL, 3d and 6d AW, and NBL) were fixed with 4% formaldehyde for 30 min. The fixed worms were embedded in paraffin and cut into a 2-μm thick cross-section with a microtome. Expression and tissue location of native TsSP1.1 in diverse stage worms was observed using IIFA. Complete worms and their cross-sections were blocked with 5% normal goat serum for 2 h, and then incubated at 37 °C for 2 h with 1:10 diluted various sera. Following washes with PBS, the worms were stained by goat anti-mouse IgG-FITC conjugate (1:100; Santa Cruz, USA). After washes again, complete worms and cross-sections were observed under fluorescence microscopy (11 (link)).
Protocol detail
Immunolocalization of TsSP1.1 in T. spiralis Stages
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Anti igg
Embedded paraffin
Fitc
Fluorescence microscopy
Formaldehyde
Goat
Microtome
Mouse
Sera
Tissue
Worms
Corresponding Organization :
Other organizations : Zhengzhou University
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Variable analysis
independent variables
- Developmental stages of T. spiralis worms (ML, 6h and 12h IIL, 3d and 6d AW, and NBL)
- Expression and tissue location of native TsSP1.1 in diverse stage worms
- Fixation of worms with 4% formaldehyde for 30 min
- Embedding of fixed worms in paraffin
- Cutting of worms into 2-μm thick cross-sections using a microtome
- Blocking of complete worms and their cross-sections with 5% normal goat serum for 2 h
- Incubation of worms with 1:10 diluted various sera at 37 °C for 2 h
- Staining of worms with goat anti-mouse IgG-FITC conjugate (1:100)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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