Home › Immunology › Use of Anti-phospho-girdin Antibodies to Visualize Intestinal Tuft Cells in Free-Floating Mouse Jejunum Cryosections
Immunology JoVE (Open Access) Citable · DOI

Use of Anti-phospho-girdin Antibodies to Visualize Intestinal Tuft Cells in Free-Floating Mouse Jejunum Cryosections

DOI: 10.3791/57475-v
What you'll learn
  • ✓ Dissect and isolate mouse jejunum tissue for immunofluorescence analysis
  • ✓ Perform immunofluorescent staining on free-floating cryosections using anti-phospho-girdin antibodies
  • ✓ Visualize and identify intestinal tuft cells via fluorescence microscopy
Protocol

Biopharma Insights Kuga et al. discovered that phosphorylation-status specific antibodies against the actin binding protein girdin phosphorylated at tyrosine 1798 (pY1798) can be used to label tuft cells (TCs). This protocol allows robust visualization of TCs using immunofluorescent staining of free-floating jejunum cryosections with pY1798 antibodies.

Difficulty
intermediate
Total time
~3-4 hours per mouse (including cryosectioning, immunostaining, and imaging)
Model organism
Mouse
Biosafety
BSL-1

Steps

1
Dissect and isolate mouse jejunum segment

Remove and isolate the jejunum tissue from euthanized mouse small intestine. Prepare tissue for cryosectioning by appropriate handling and preservation.

▶ 00:40
2
Perform immunofluorescence on free-floating cryosections

Incubate cryosections with anti-phospho-girdin (pY1798) antibodies and appropriate secondary fluorescent antibodies to label tuft cells specifically.

▶ 02:40
3
Acquire and analyze fluorescence microscopy images

Image stained jejunum cryosections using fluorescence microscopy to visualize and characterize labeled intestinal tuft cells.

▶ 04:42
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