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Image it fx reagent

Manufactured by Thermo Fisher Scientific

The Image-iT FX reagent is a fluorescence-based labeling solution used for protein visualization in biological samples. It provides a simple and efficient method for labeling and detecting specific proteins in cells or tissues.

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4 protocols using image it fx reagent

1

Histological Analysis of Tissue Samples

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Paraffin embedding and sectioning was performed by the Moores cancer center histology core at UCSD. For OCT embedding, tissue biopsies were first fixed with 4% PFA for 2 hours then cryoprotected by sucrose for 48 hours prior to being embedded in OCT compound and stored in −80 ⁰C. Histological analysis was performed using either paraffin sections or frozen sections by Hematoxylin-Eosin (H&E) staining. Collagen was stained by the Richard-Allan Scientific Gomori Trichrome green collagen staining kit (ThermoFisher Scientific, Waltham, MA, USA). Elastic staining (Thermo Fisher Scientific, Waltham, MA, USA) and hydroxyproline assay (BioVision) were performed on human or mouse tissues according to manufacturer’s instructions. For IHC, fixed and permeabilized tissue sections were blocked with Image-iT FX reagent (invitrogen) before incubating with primary antibodies followed by appropriate 488- or Cy3-coupled secondary antibodies. Nuclei were counter-stained with DAPI. All images were taken with an Olympus BX41 microscope (widefield) or Zeiss LSM510 confocal microscope as indicated.
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2

Multimodal Tissue Analysis Protocol

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Tissue biopsies were directly embedded in the OCT compound or fixed with Carnoy’s fluid. Paraffin-embedded tissues were used for hematoxylin/eosin (H&E) staining, and frozen sections were fixed in 4% PFA for 20 min to immunofluorescence staining. For Gram staining, tissue is fixed with Carnoy’s fluid and embedded in OCT compound, sectioned by 4um and stained with Gram stain (Remel, Lenexa, KS). For Periodic acid-Schiff(PAS) staining, tissue is fixed with Carnoy’s fluid and embedded in Paraffin, and stained with PAS. For TUNEL staining, TUNEL Andy Fluo 594 Apoptosis Detection Kit (#A051, abpbio) is used. For IHC, fixed and permeabilized frozen tissue sections were blocked with Image-iT FX reagent (Invitrogen) before incubating with HABP (#385911, EMD Millipore), Muc2 (#PIMA512345, Fisher) or Reg3g (#PA5-50450, Thermo Fisher) in 1:100 dilution followed by appropriate 488- or 568-coupled secondary antibodies. Nuclei were counterstained with DAPI. All images were taken with an Olympus BX41 microscope (widefield) or Zeiss LSM510 confocal microscope as indicated.
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3

Histological Analysis of Tissue Samples

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Tissue biopsies were directly embedded in OCT compound or paraffin. Paraffin embedded tissues are used for Hematoxylin/Eosin (H&E) staining, and frozen sections were fixed in 4% PFA for 20 mins or 100% acetone prior to immunofluorescence staining. For IHC, fixed and permeabilized frozen tissue sections were blocked with Image-iT FX reagent (Invitrogen) before incubating with primary antibodies followed by appropriate 488- or 568-coupled secondary antibodies. Nuclei were counterstained with DAPI. All images were taken with an Olympus BX41 microscope (widefield) or Zeiss LSM510 confocal microscope as indicated.
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4

Histological Analysis of Tissue Samples

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Tissue biopsies were directly embedded in OCT compound or paraffin. Paraffin embedded tissues are used for Hematoxylin/Eosin (H&E) staining, and frozen sections were fixed in 4% PFA for 20 mins or 100% acetone prior to immunofluorescence staining. For IHC, fixed and permeabilized frozen tissue sections were blocked with Image-iT FX reagent (Invitrogen) before incubating with primary antibodies followed by appropriate 488- or 568-coupled secondary antibodies. Nuclei were counterstained with DAPI. All images were taken with an Olympus BX41 microscope (widefield) or Zeiss LSM510 confocal microscope as indicated.
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