Immunoenzyme Multiple Staining Methods

Noorden, Van

doi:10.1046/j.1525-1314.2002.00206.x-i1

Cited by 1 publication

(1 citation statement)

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“…IHC is commonly used for colocalization, most frequently by using two different antibodies conjugated in different host species and visualized with different colored chromogens . However, using multiple chromogens can lead to nonspecific background staining or incompatibility of colors 11,12 .…”

Section: Introductionmentioning

confidence: 99%

Sequential Immunofluorescence and Immunohistochemistry on Cryosectioned Zebrafish Embryos

Ferguson¹,

Shive²

2019

JoVE

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Investigation of intercellular interactions often requires discrete labeling of specific cell populations and precise protein localization. The zebrafish embryo is an excellent tool for examining such interactions with an in vivo model. Whole-mount immunohistochemical and immunofluorescence assays are frequently applied in zebrafish embryos to assess protein expression. However, it can be difficult to achieve accurate mapping of co-localized proteins in three-dimensional space. In addition, some studies may require the use of two antibodies that are not compatible with the same technique (e.g., antibody 1 is only suitable for immunohistochemistry and antibody 2 is only suitable for immunofluorescence). The purpose of the method described herein is to perform sequential immunofluorescence and/or immunohistochemistry on individual cryosections derived from early-stage zebrafish embryos. Here we describe the use of sequential rounds of immunofluorescence, imaging, immunohistochemistry, imaging for a single cryosection in order to achieve precise identification of protein expression at the single-cell level. This methodology is suitable for any study in early-stage zebrafish embryos that requires accurate identification of multiple protein targets in individual cells.

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Section: Introductionmentioning

confidence: 99%